Objective To investigate the effects and underlying mechanisms of total flavonoids of sarcandra glabra(TFFSG)on bone marrow mesenchymal stem cells(BMSCs)and their derived exosomes in immune thrombo-cytopenia(ITP),with a focus on promoting megakaryocyte differentiation and maturation.Methods BMSCs induced by rabbit anti-rat platelet serum(APS)were divided into five groups:a blank control group,an ITP-BMSCs model group,and three TFFSG intervention groups with low(1.95 μg/mL),medium(3.90 μg/mL),and high doses(7.80 μg/mL).The apoptosis rates and the expression levels of apoptosis-related proteins-B-cell lymphoma 2(Bcl-2),Bcl-2-associated X protein(BAX),and Cysteinyl aspartate-specific proteinase-3(Caspase-3)-were assessed.Exosomes were isolated from the blank control group(NC-BMSCs-Exos),the ITP-BMSCs model group(ITP-BMSCs-Exos),and the medium-dose TFFSG group(TFFSG-BMSCs-Exos).Each group's exosomes(5 μg/mL)were co-cultured with megakaryocytic lineage Dami cells for 96 hours.Flow cytometry was employed to evaluate the expression of megakaryocytic differentiation markers(CD41a,CD42b,CD61)and the proportion of polyploid cells(≥4 N)in each group.Western Blot analysis was conducted to examine the expression of p-MEK1/2,MEK1/2,p-ERK1/2,and ERK1/2 across all groups.Results Compared with the ITP-BMSCs model group,the apoptosis rates in all TFFSG intervention groups were significantly reduced(P<0.01).In the medium-and high-dose TFFSG groups,BAX and Caspase-3 expression levels were markedly downregulated,whereas Bcl-2 expression was upregulated(P<0.05,P<0.01).Compared with the ITP-BMSCs-Exos group,the TFFSG-BMSCs-Exos group demonstrated increased expression of CD41a+,CD42b+,and CD61+,a higher proportion of polyploid cells(≥4 N)(P<0.05),as well as elevated ratios of p-MEK1/2 to MEK1/2 and p-ERK1/2 to ERK1/2(P<0.05).Conclusion TFFSG inhibits apopto-sis of ITP-state BMSCs in vitro and promotes megakaryocyte differentiation and polyploidization maturation through BMSC-derived exosomes by activating the MEK1/2-ERK1/2 signaling pathway.

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 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 investigate the effects and underlying mechanisms of total flavonoids of sarcandra glabra(TFFSG)on bone marrow mesenchymal stem cells(BMSCs)and their derived exosomes in immune thrombo-cytopenia(ITP),with a focus on promoting megakaryocyte differentiation and maturation.Methods BMSCs induced by rabbit anti-rat platelet serum(APS)were divided into five groups:a blank control group,an ITP-BMSCs model group,and three TFFSG intervention groups with low(1.95 μg/mL),medium(3.90 μg/mL),and high doses(7.80 μg/mL).The apoptosis rates and the expression levels of apoptosis-related proteins-B-cell lymphoma 2(Bcl-2),Bcl-2-associated X protein(BAX),and Cysteinyl aspartate-specific proteinase-3(Caspase-3)-were assessed.Exosomes were isolated from the blank control group(NC-BMSCs-Exos),the ITP-BMSCs model group(ITP-BMSCs-Exos),and the medium-dose TFFSG group(TFFSG-BMSCs-Exos).Each group's exosomes(5 μg/mL)were co-cultured with megakaryocytic lineage Dami cells for 96 hours.Flow cytometry was employed to evaluate the expression of megakaryocytic differentiation markers(CD41a,CD42b,CD61)and the proportion of polyploid cells(≥4 N)in each group.Western Blot analysis was conducted to examine the expression of p-MEK1/2,MEK1/2,p-ERK1/2,and ERK1/2 across all groups.Results Compared with the ITP-BMSCs model group,the apoptosis rates in all TFFSG intervention groups were significantly reduced(P<0.01).In the medium-and high-dose TFFSG groups,BAX and Caspase-3 expression levels were markedly downregulated,whereas Bcl-2 expression was upregulated(P<0.05,P<0.01).Compared with the ITP-BMSCs-Exos group,the TFFSG-BMSCs-Exos group demonstrated increased expression of CD41a+,CD42b+,and CD61+,a higher proportion of polyploid cells(≥4 N)(P<0.05),as well as elevated ratios of p-MEK1/2 to MEK1/2 and p-ERK1/2 to ERK1/2(P<0.05).Conclusion TFFSG inhibits apopto-sis of ITP-state BMSCs in vitro and promotes megakaryocyte differentiation and polyploidization maturation through BMSC-derived exosomes by activating the MEK1/2-ERK1/2 signaling pathway.

Objective:To explore the association of frailty and serum C-terminal agrin fragment(CAF)with the prognosis of elderly patients with acute coronary syndrome(ACS).Methods:In this prospective cohort study, clinical data of 207 older patients with ACS between January 2020 and May 2022 were collected.Serum samples were obtained within 24 hours after enrollment to detect CAF levels.Meanwhile, the thrombolysis in myocardial infarction(TIMI)and frailty screening questionnaire(FSQ)scores were assessed on admission.Patients were followed up for major adverse cardiovascular and cerebrovascular events(MACCE)for 90 days.Multivariate logistic regression was used to analyze the influencing factors of MACCE.The receiver operating characteristic(ROC)curve was performed to evaluate the predictive ability of the FSQ score, serum CAF and their combination for MACCE.According to 90-day mortality, patients were divided into a survival group(n=176)and a death group(n=31). The Cox proportional hazards regression model was used for survival analysis.Results:The FSQ score( Z=4.412, P<0.001)and serum CAF( Z=6.702, P<0.001)in the MACCE group were higher than those in the non-MACCE group.Logistic regression analysis showed that after adjusting for age, sex, TIMI score and complete revascularization, frailty defined by FSQ( OR=1.714; 95% CI: 1.059-2.775; P=0.028)and high serum CAF( OR=1.230; 95% CI: 1.122-1.350; P<0.05)were independent risk factors for MACCE.The area under the ROC curve(AUC)of the FSQ score for predicting MACCE was 0.797(95% CI: 0.735-0.850; P<0.001), the predictive cut-off point was an FSQ score >2, and the Youden index(YI)was 0.419, yielding a sensitivity of 0.708 and a specificity of 0.711.In addition, the AUC of serum CAF for predicting MACCE was 0.766(95% CI: 0.701-0.822; P<0.001), the predictive cut-off point was >6.01 μg/L, and YI was 0.460, yielding a sensitivity of 0.750 and a specificity of 0.710.The predictive ability of FSQ combined with CAF for MACCE was higher than FSQ( Z=2.294, P=0.022)or CAF( Z=2.545, P=0.011)alone.Cox regression analysis showed that frailty defined by FSQ( HR=3.487; 95% CI: 1.329-9.153; P=0.011)was independently associated with all-cause mortality within 90 days after ACS. Conclusions:Frailty assessment and serum CAF detection can improve the risk stratification of elderly patients with ACS.

Objective To screen the active components of total flavonoid extracts of Sarcandra glabra to promote megakaryocyte differentiation.Methods(1)A model of megakaryocyte differentiation disorder was established by co-culturing human megakaryocytic leukaemia cells(Dami)with human bone marrow stromal cells(HS-5)as an evaluation system,and the experimental groupings were as follows:the Dami group(Dami),the control group(Dami+HS-5),and the PMA group[Dami+HS-5+5 ng·mL-1 foprolol 12-tetradecanoate 13-acetate(PMA)],and model group[Dami+HS-5+1%rabbit anti-rat platelet serum(APS)+5 ng·mL-1 PMA]were cultured for 48 hours.The expressions of megakaryocyte differentiation and maturation surface marker molecules,CD41a and CD61 were detected by flow cytometry.(2)Forty-nine SD male rats were randomly divided into blank plasma group,15-minute group,30-minute group,60-minute group,90-minute group,120-minute group,and 240-minute group,with 7 rats in each group.The rats in each administration group were gavaged with 1.26 g·kg-1 of total flavonoids extracts of Sarcandra glabra,and blood was collected at six set time points(15,30,60,90,120,240 minutes)for the preparation of time-dependent serum-containing plasma of total flavonoids extracts of Sarcandra glabra.(3)Ultra-high performance liquid chromatography-quadrupole tandem time-of-flight mass spectrometry(UHPLC-Q-TOF/MS)was used to analyze the plasma of the time-dependent serum-containing plasma of the total flavonoids extracts of Sarcandra glabra,and the peak area was used to construct a matrix(X-matrix)of the amount of chemical composition change over time in the time-dependent serum-containing plasma of the total flavonoids extracts of Sarcandra glabra.The collected time-dependent serum-containing plasma of the total flavonoids extracts of Sarcandra glabra at six different time points was used to intervene in the model of megakaryocyte differentiation and maturation disorder,and the expression of cell surface molecules CD41a and CD61 was detected by flow cytometry to construct the matrix of effect of time-dependent serum-containing plasma of the total flavonoids extracts of Sarcandra glabra(Y-matrix).(4)After the data of X and Y matrices were standardized,partial least squares(PLS)was used to calculate and analyze the quantitative and qualitative effect relationship,and variable importance for projection(VIP)>1 was used as the threshold to screen the effect components related to the changes of cell surface molecules CD41a and CD61,and chemical composition identification,as the potential effector components in the total flavonoid extracts of Sarcandra glabra were used to promote the differentiation of megakaryocytes,and finally the regression evaluation system was used to verify the efficacy of its medicinal effect.Results(1)Compared with the Dami group,the expression level of CD41a on the surface of Dami cells in the control group was significantly increased(P＜0.05).Compared with the control group,the expression levels of CD41a and CD61 on the surface of Dami cells in the PMA group were significantly increased(P＜0.01).Compared with the PMA group,the expression levels of CD41a and CD61 on the surface of Dami cells in the model group were significantly reduced(P＜0.01).(2)Compared with the blank plasma group,the expression levels of the molecules CD41a and CD61 on the surface of Dami cells at each time point of 15,30,60,90,120,and 240 minutes were significantly increased(P＜0.01),and the expression levels of CD41a and CD61 were both highest in the 30-minute group.The potential effective components with VIP value greater than 1 were screened out in the positive and negative ion mode,and 540.3638@12.25 and 559.2991@11.53 were selected for pharmacodynamic verification.559.2991@11.53 was identified as daucosterol(Dau),540.3638@12.25 was identified as rosmarinic acid 4-O-β-D-glucoside(Ros).After Ros and Dau intervened in the megakaryocyte differentiation and maturation disorder model respectively,the expression levels of CD41a and CD61 on the surface of Dami cells in the low-,medium-and high-dose groups(40,60 and 80 μg·mL-1)of Ros and Dau were significantly increased compared with the model group(P＜0.05,P＜0.01).Conclusion Ros and Dau may be the active components of the total flavonoids extracts of Sarcandra glabra to promote the differentiation of megakaryocytes.

Objective:To examine the prevalence of frailty among elderly patients in the emergency department and to investigate the potential relationship between vitamin D nutritional status and frailty.Methods:This study collected clinical data from elderly patients aged over 65 years in the emergency intensive care unit and emergency observation ward of Beijing Bo'Ai Hospital from January to September 2021.The data included blood routine, biochemical indicators, circulating interleukin-6, cortisol, thyrotropin, and 25-hydroxyvitamin D[25(OH)D], which were detected within 24 hours after enrollment.Additionally, the Frailty Screening Questionnaire(FSQ), FRAIL scale, and Clinical Frailty Scale(CFS)were used to score the patients.Based on the scores, the patients were divided into frail or non-frail groups, and the prevalence of frailty was reported accordingly using the criteria of the aforementioned scales.The consistency of the three scales was evaluated using the Spearman rank test and Kappa coefficient.We compared the differences in clinical data and laboratory indicators of patients between the frail and non-frail groups.Additionally, we used a multivariable Logistic regression model to analyze the association between vitamin D nutritional status and frailty.We also analyzed the prevalence of frailty in different vitamin D nutritional statuses and evaluated the predictive ability of serum 25(OH)D for frailty using the receiver operating characteristic(ROC)curve.Results:A total of 317 patients were included in the study.The prevalence of frailty in elderly patients in the emergency department was found to be 47.0%, 55.2%, and 69.4% according to the FSQ, FRAIL, and CFS scales, respectively.The study evaluated the consistency of these three scales, revealing a Spearman rank correlation coefficient of 0.761(95% CI: 0.715-0.806, P<0.001)and a Kappa coefficient of 0.536(95% CI: 0.451-0.621, P<0.001)between FSQ and FRAIL, which were the highest correlations observed.Logistic regression analysis, after adjusting for age, gender, BMI, and other factors, indicated that vitamin D deficiency( OR=5.994, 95% CI: 1.232-29.169, P=0.027)was independently associated with an increased prevalence of frailty as defined by FSQ criteria.The prevalence of frailty increased with the severity of vitamin D malnutrition.In the vitamin D deficiency group, the prevalence was higher compared to the vitamin D insufficiency and sufficiency groups( P<0.05 for all).The area under the ROC curves(AUCs)of serum 25(OH)D levels to predict frailty, as defined by FSQ, FRAIL, and CFS, were 0.806(95% CI: 0.744-0.868), 0.748(95% CI: 0.679-0.817), and 0.768(95% CI: 0.701-0.826)( P<0.001 for all).The optimal cut-off values were 12.0, 9.76, and 11.65 μg/L, respectively, yielding a Youden index of 0.553, 0.419, and 0.462. Conclusions:FSQ, FRAIL, and CFS demonstrated a strong level of consistency in assessing frailty.Additionally, serum 25(OH)D can serve as an independent predictor of frailty, aiding in the identification of frail individuals and enhancing the risk stratification of elderly patients in the emergency department.

Objective:To investigate the association between serum interleukin (IL) -6 and silent information regulator (SIRT) -1 and frailty in elderly patients in the emergency department.Methods:This was a cross-sectional study. Patients aged 60 years and above treated in the emergency department of Beijing Bo'Ai Hospital from January to December 2022 were collected. Blood routine, biochemical indicators, and serum IL-6 were detected within 24 h after enrollment. At the same time, fasting venous blood 2 mL was collected and the serum was stored at minus 80℃ after centrifugation. The level of SIRT-1 was detected by enzyme-linked immunosorbent assay. Nutritional risk screening 2002 was performed within 72 h, Barthel index was used to assess the ability of daily living and grip strength was measured. The patients were divided into frailty and non-frailty groups according to Fried frailty phenotype (FP). The differences of clinical data and laboratory indicators were compared between the two groups. Multivariable logistic regression model was used to analyze the association between serum IL-6, SIRT-1 and frailty. The predictive ability of serum IL-6 and SIRT-1 for frailty was evaluated by the receiver operating characteristic (ROC) curve.Results:A total of 316 elderly patients in the emergency department were included in this study and divided into frailty group ( n=156) and non-frailty group ( n=160) according to Fried FP criteria. Univariate analysis showed that serum IL-6 [33.3 (13.0, 69.2) ng/L vs. 20.0 (9.2, 41.3) ng/L, P=0.001] and SIRT-1 [(9.98±1.23) μg/L vs. (8.98±1.65) μg/L, P<0.001] of patients in the frailty group were higher than those in the non-frailty group. Multivariable logistic regression analysis showed that serum IL-6 ( OR=1.006, 95% CI: 1.001-1.011, P=0.036) and SIRT-1 ( OR=1.838, 95% CI: 1.475-2.290, P<0.001) were independently associated with frailty after adjusting for age, sex, body mass index, Barthel index and grip strength. The area under the curve (AUC) of serum IL-6 for predicting frailty was 0.671 (95% CI: 0.604-0.738, P<0.001), the predictive cut-off point was 33.8 ng/L. The AUC of SIRT-1 for predicting frailty was 0.736 (95% CI: 0.674-0.799, P<0.001), the predictive cut-off point was 9.13 μg/L. The AUC of the model of IL-6 combined with SIRT-1 was 0.765 (95% CI: 0.707-0.823, P<0.001), the sensitivity and specificity were 0.776 and 0.726, respectively, and its predictive efficacy was superior to that of IL-6 alone ( Z=2.119, P=0.034). Conclusion:Serum IL-6 and SIRT-1 are independent predictors of frailty in elderly patients in the emergency department.

Objective:To analyze the association between the transverse area of erector spine muscle at T 12 level and prognosis in elderly patients with acute coronary syndrome (ACS). Methods:This was a prospective cohort study. Clinical data of elderly patients with ACS admitted to the coronary care unit or the emergency intensive care unit in Beijing Bo'Ai Hospital from January 2019 to June 2022. Blood routine and biochemical indicators were detected and echocardiography was performed within 24 hours after admission, chest CT examination was completed and the cross-sectional areas of the left and right vertical spinalis muscles were calculated at T 12 level, thrombolysis in myocardial infarction (TIMI) was performed within 24 hours after admission. The occurrence of major adverse cardiovascular and cerebral events (MACCE) within 12 months was followed up. The differences of clinical data and laboratory indexes between the MACCE group and the non-MACCE group were compared. Multivariable logistic regression model was used to analyze the association between the cross-sectional area of vertical spinal muscle at T 12 level and the occurrence of MACCE within 12 months after ACS. Results:A total of 406 patients were enrolled in the study, the incidence of MACCE within 12 months was 22.2%. The cross-sectional area of erector spine muscle at T 12 level [(20.43±3.51) cm 2vs. (24.36±3.66) cm 2, (Z=4.412, P<0.001)] in the MACCE group was lower than that in the non-MACCE group. Logistic regression analysis showed that after adjusting for sex, age, body mass index (BMI), TIMI score and revascularization, the cross-sectional area of erector spine muscle at T 12 level ( OR = 0.883, 95% CI: 0.803-0.971, P=0.010) was independently associated with MACCE within 12 months after ACS. Conclusion:The transverse area of vertical spinal muscle at T 12 level can be used as an independent predictor of MACCE within 12 months in elderly patients with ACS.

ObjectiveTo investigate the association between serum 25-hydroxy vitamin D [25(OH)D] and the occurrence and outcome of stroke-associated pneumonia (SAP) in patients with acute ischemic stroke (AIS) in emergency ward. MethodsThe clinical data of 256 patients with AIS from January, 2019 to December, 2021 were collected in the emergency department of Beijing Bo'ai Hospital. Blood routine, biochemical indicators and serum concentration of 25(OH)D were detected within 24 hours after enrollment; meanwhile, National Institute of Health Stroke Scale (NIHSS) and A2DS2 score were evaluated. The patients were divided into non-SAP group (n = 164) and SAP group (n = 92) according to whether pneumonia occurred during hospitalization. Multivariable logistic regression model was used to analyze the influencing factors of SAP. The predictive ability of serum 25(OH)D and A2DS2 for SAP were evaluated by receiver operating characteristic (ROC) curves. The 28-day survival of patients with SAP was followed up. Multivariable Cox proportional hazard regression model was used to investigate the association between vitamin D nutritional status and 28-day all-cause mortality. ResultsSerum 25(OH)D was significantly lower in the SAP group than that in the non-SAP group (Z = 6.896, P < 0.001). After adjusting age, sex, infarct volume, A2DS2 score and other factors, lower serum 25(OH)D level (OR = 0.934, 95%CI 0.884 to 0.986, P = 0.014) was an independent risk factor for SAP. The areas under curve (95%CI) of serum 25(OH)D, A2DS2 score and their combined model for predicting SAP were 0.774 (0.718 to 0.824), 0.832 (0.781 to 0.876) and 0.851 (0.802 to 0.893) (P < 0.001), respectively; and the optimum cut-off values were 25(OH)D < 10.2 ng/mL, A2DS2 score > 5 points, combined prediction > 0.207, and the Youden index were 0.493, 0.662 and 0.616, respectively. A2DS2 score could improve the prediction efficiency of serum 25(OH)D (Z = 2.106, P = 0.035). After adjusting age, sex, infarct volume and NIHSS score, vitamin D deficiency was an independent risk factor for all-cause mortality after 28 days of SAP (HR = 2.871, 95%CI 1.004 to 8.208, P = 0.049) . ConclusionSerum 25(OH)D is independently associated with the occurrence and outcome of SAP in patients with AIS in emergency ward, which could serve as an independent predictor for SAP.