Objective:To investigate the effect of PD-1/PD-L1 inhibitor BMS-1 on LPS-induced inflammation in mouse microg-lia cells(BV-2 cells).Methods:Bv-2 cells were divided into Control group,LPS group and LPS+BMS-1 group.Bv-2 cells in Control group were cultured in DMEM medium for 78 hours,cells in LPS group were stimulated with 100 ng/ml LPS for 6 hours after 72 hours of normal culture,Bv-2 cells in LPS+BMS-1 group were treated with 50 nmol/ml BMS-1 for 72 hours and then stimulated with 100 ng/ml LPS for 6 hours.Expressions of PD-1 and iNOS mRNA in each group were detected by RT-qPCR,and expressions of PD-1 and iNOS protein in microglia were detected by Western blot.Flow cytometry was used to detect cell apoptosis in each group.Levels of inflamma-tory cytokines IL-1β,IL-6,TNF-α and IL-10 were detected by ELISA.Results:RT-qPCR and Western blot results showed that com-pared with Control group,LPS group had significantly increased expression of PD-1 and iNOS(P<0.05).Compared with LPS group,LPS+BMS-1 group had significantly decreased expression of PD-1(P<0.05)and significantly increased expression of iNOS(P<0.05).Flow cytometry showed that compared with Control group,LPS group had a significantly increased in apoptosis of microglia(P<0.000 1).Compared with LPS group,LPS+BMS-1 group had a significantly increased in apoptosis of microglia(P<0.000 1).ELISA results showed that compared with Control group,LPS group had no significantly increased in pro-inflammatory factors IL-1β and IL-6(P>0.05),while significantly increased in TNF-α(P<0.000 1)and anti-inflammatory factor IL-10(P<0.000 1).Pro-inflammatory cyto-kine IL-1β in LPS+BMS-1 group was significantly higher than that in LPS group(P=0.000 1),IL-6 and TNF-α were also significantly higher than those in LPS group(P<0.000 1),while anti-inflammatory cytokine IL-10 in LPS+BMS-1 group was significantly lower than that in LPS group(P<0.000 1).Conclusion:BMS-1 can promote LPS-induced inflammatory response or impede the recovery of inflammation,and increase apoptosis of microglia.PD-1/PD-L1 pathway may be a potential therapeutic target for neuroinflammation.

Objective The aim of this study was to detect the expression of miR-383-5p in bladder cancer tissues and bladder cancer 5637 cells,BIU-87 cells,TCCSUP cells and HT-1376 cells,and to explore the effects of miR-383-5p on the prolif-eration,invasion,migration and apoptosis of bladder cancer cells by targeting CIP2A.Methods The expression of miR-383-5p was detected by qRT-PCR in human bladder cancer tissues and their corresponding adjacent tissues,5637 cells,BIU-87 cells,TCCSUP cells,HT-1376 cells,human bladder transitional epithelial cells.BIU-87 cells with low miR-383-5p expression were selected for subsequent experiments.BIU-87 cells were divided into the blank group(normal culture),miR-383-5p NC group(negative control,transfected with miR-383-5p negative control),miR-383-5p mimic group(transfected with miR-383-5p mimic),and miR-383-5p mimic+pc-CIP2A group(co-transfected with miR-383-5p mimic and CIP2A overexpression plasmid pc-CIP2A).CCK-8 kit was used to detect the viability of BIU-87 cells in each group;Flow cytometry was used to detect apoptosis of BIU-87 cells;Transwell assay was used to measure cell invasion ability of BIU-87 cells;Scratch assay was used to measure cell migration ability of BIU-87 cells;Western blot was used to determine the expression of proteins related to apoptosis,invasion(MMP-2,MMP-9),and CIP2A/PP2A in BIU-87 cells;The dual luciferase assay was used to verify the targeting relationship between miR-383-5p and CIP2A in BIU-87 cells.Results The expression of miR-383-5p was low in bladder cancer tissues and bladder cancer cells.Compared with the blank group,BIU-87 cells in the miR-383-5p mimic group showed a significant increase the level of miR-383-5p(0.91±0.10 vs.1.67±0.24,P<0.01)and a significant decrease in the expression of CIP2A protein(1.32±0.17 vs.0.45±0.03,P<0.001),the cell viability,invasion,migration abilities,the expression of proteins related to invasion(MMP-2,MMP-9),and the expression of Bcl-2 protein[(100.00±4.36)% vs.(32.15±2.65)% ,(150.20±12.95)vs.(82.35±7.01),(77.91±3.63)% vs.(46.12±2.54)% ,1.02±0.11 vs.0.22±0.04,1.03±0.18 vs.0.21±0.04,1.01±0.14 vs.0.27±0.05,P<0.001];The apoptosis rate,the expression of caspase-3 and Bax proteins related to apoptosis,and PP2A expression were significantly increased[(14.02±2.29)% vs.(38.21±3.20)% ],0.81±0.11 vs.1.78±0.24,0.83±0.12 vs.1.72±0.24,0.27±0.02 vs.0.95±0.16,P<0.001].Compared with the miR-383-5p mimic group,BIU-87 cells in the miR-383-5p mimic+pc-CIP2A group significantly increased the cell viability,invasion,migration abilities,the expression of proteins related to invasion,and the expression of Bcl-2 protein[(32.15±2.65)% vs.(50.18±3.77)% ,(82.35±7.01)% vs.(116.30±13.70),(46.12±2.54)% vs.(58.43±3.15)% ,0.22±0.04 vs.0.60±0.08,0.21±0.04 vs.0.5 8±0.06,0.27±0.05 vs.0.64±0.08,P<0.05];The apoptosis rate,the expression of caspase-3,Bax,and PP2A was signifi-cantly reduced in the miR-383-5p mimic+pc-CIP2A group[(38.21±3.20)% (23.15±2.74)% ,1.78±0.24 vs.1.25±0.21,1.72±0.24 vs.1.23±0.18,0.95±0.16 vs.0.60±0.13,P<0.05].The results of dual luciferase experiments showed a corresponding tar-geting relationship between miR-383-5p and CIP2A.Conclusion Increasing the expression of miR-383-5p can inhibit the prolif-eration,invasion and migration of bladder cancer BIU-87 cells,and enhance the ability of apoptosis,which may be achieved by targe-ted regulation of CIP2A.

Objective:To evaluate the association between heatwaves and fall-related mortality.Methods:A total of 61 421 fall-related mortality from 2013 to 2022 in 7 provinces of China were included in a time-stratified case-crossover design, with daily meteorological data derived from the fifth generation European Reanalysis dataset produced by the European Centre for Medium-Range Weather Forecasts. Conditional logistic regression chimeric distributed lag nonlinear model was used to analyze the association between heatwaves and fall-related mortality and stratified analysis was conducted according to gender and age.Results:Heatwaves were associated with an increased risk of fall-related morality. The risk of fall-related mortality during heatwaves was higher than during non-heatwave periods ( OR=1.11, 95% CI: 1.05-1.18). The attributable fraction of fall-related motality due to heatwaves was 10.25% (95% CI: 4.49%-15.36%). For each 1 ℃ increase above the heatwave threshold, the risk of fall-related mortality increased by 34% ( OR=1.34, 95% CI: 1.02-1.76). The effect of heatwave duration on fall-related mortality was not statistically significant. Stratified analyses indicated that women experienced a higher risk of fall-related mortality during heatwaves ( OR=1.13, 95% CI: 1.04-1.22) compared to man ( OR=1.10, 95% CI: 1.04-1.17). Conclusions:Heatwave increases the risk of fall-related mortality, and the intensity of heatwaves modify this risk. Women are vulnerable populations.

Objective:To evaluate the association between heatwaves and fall-related mortality.Methods:A total of 61 421 fall-related mortality from 2013 to 2022 in 7 provinces of China were included in a time-stratified case-crossover design, with daily meteorological data derived from the fifth generation European Reanalysis dataset produced by the European Centre for Medium-Range Weather Forecasts. Conditional logistic regression chimeric distributed lag nonlinear model was used to analyze the association between heatwaves and fall-related mortality and stratified analysis was conducted according to gender and age.Results:Heatwaves were associated with an increased risk of fall-related morality. The risk of fall-related mortality during heatwaves was higher than during non-heatwave periods ( OR=1.11, 95% CI: 1.05-1.18). The attributable fraction of fall-related motality due to heatwaves was 10.25% (95% CI: 4.49%-15.36%). For each 1 ℃ increase above the heatwave threshold, the risk of fall-related mortality increased by 34% ( OR=1.34, 95% CI: 1.02-1.76). The effect of heatwave duration on fall-related mortality was not statistically significant. Stratified analyses indicated that women experienced a higher risk of fall-related mortality during heatwaves ( OR=1.13, 95% CI: 1.04-1.22) compared to man ( OR=1.10, 95% CI: 1.04-1.17). Conclusions:Heatwave increases the risk of fall-related mortality, and the intensity of heatwaves modify this risk. Women are vulnerable populations.

Objective:To investigate the effect of PD-1/PD-L1 inhibitor BMS-1 on LPS-induced inflammation in mouse microg-lia cells(BV-2 cells).Methods:Bv-2 cells were divided into Control group,LPS group and LPS+BMS-1 group.Bv-2 cells in Control group were cultured in DMEM medium for 78 hours,cells in LPS group were stimulated with 100 ng/ml LPS for 6 hours after 72 hours of normal culture,Bv-2 cells in LPS+BMS-1 group were treated with 50 nmol/ml BMS-1 for 72 hours and then stimulated with 100 ng/ml LPS for 6 hours.Expressions of PD-1 and iNOS mRNA in each group were detected by RT-qPCR,and expressions of PD-1 and iNOS protein in microglia were detected by Western blot.Flow cytometry was used to detect cell apoptosis in each group.Levels of inflamma-tory cytokines IL-1β,IL-6,TNF-α and IL-10 were detected by ELISA.Results:RT-qPCR and Western blot results showed that com-pared with Control group,LPS group had significantly increased expression of PD-1 and iNOS(P<0.05).Compared with LPS group,LPS+BMS-1 group had significantly decreased expression of PD-1(P<0.05)and significantly increased expression of iNOS(P<0.05).Flow cytometry showed that compared with Control group,LPS group had a significantly increased in apoptosis of microglia(P<0.000 1).Compared with LPS group,LPS+BMS-1 group had a significantly increased in apoptosis of microglia(P<0.000 1).ELISA results showed that compared with Control group,LPS group had no significantly increased in pro-inflammatory factors IL-1β and IL-6(P>0.05),while significantly increased in TNF-α(P<0.000 1)and anti-inflammatory factor IL-10(P<0.000 1).Pro-inflammatory cyto-kine IL-1β in LPS+BMS-1 group was significantly higher than that in LPS group(P=0.000 1),IL-6 and TNF-α were also significantly higher than those in LPS group(P<0.000 1),while anti-inflammatory cytokine IL-10 in LPS+BMS-1 group was significantly lower than that in LPS group(P<0.000 1).Conclusion:BMS-1 can promote LPS-induced inflammatory response or impede the recovery of inflammation,and increase apoptosis of microglia.PD-1/PD-L1 pathway may be a potential therapeutic target for neuroinflammation.

Objective The aim of this study was to detect the expression of miR-383-5p in bladder cancer tissues and bladder cancer 5637 cells,BIU-87 cells,TCCSUP cells and HT-1376 cells,and to explore the effects of miR-383-5p on the prolif-eration,invasion,migration and apoptosis of bladder cancer cells by targeting CIP2A.Methods The expression of miR-383-5p was detected by qRT-PCR in human bladder cancer tissues and their corresponding adjacent tissues,5637 cells,BIU-87 cells,TCCSUP cells,HT-1376 cells,human bladder transitional epithelial cells.BIU-87 cells with low miR-383-5p expression were selected for subsequent experiments.BIU-87 cells were divided into the blank group(normal culture),miR-383-5p NC group(negative control,transfected with miR-383-5p negative control),miR-383-5p mimic group(transfected with miR-383-5p mimic),and miR-383-5p mimic+pc-CIP2A group(co-transfected with miR-383-5p mimic and CIP2A overexpression plasmid pc-CIP2A).CCK-8 kit was used to detect the viability of BIU-87 cells in each group;Flow cytometry was used to detect apoptosis of BIU-87 cells;Transwell assay was used to measure cell invasion ability of BIU-87 cells;Scratch assay was used to measure cell migration ability of BIU-87 cells;Western blot was used to determine the expression of proteins related to apoptosis,invasion(MMP-2,MMP-9),and CIP2A/PP2A in BIU-87 cells;The dual luciferase assay was used to verify the targeting relationship between miR-383-5p and CIP2A in BIU-87 cells.Results The expression of miR-383-5p was low in bladder cancer tissues and bladder cancer cells.Compared with the blank group,BIU-87 cells in the miR-383-5p mimic group showed a significant increase the level of miR-383-5p(0.91±0.10 vs.1.67±0.24,P<0.01)and a significant decrease in the expression of CIP2A protein(1.32±0.17 vs.0.45±0.03,P<0.001),the cell viability,invasion,migration abilities,the expression of proteins related to invasion(MMP-2,MMP-9),and the expression of Bcl-2 protein[(100.00±4.36)% vs.(32.15±2.65)% ,(150.20±12.95)vs.(82.35±7.01),(77.91±3.63)% vs.(46.12±2.54)% ,1.02±0.11 vs.0.22±0.04,1.03±0.18 vs.0.21±0.04,1.01±0.14 vs.0.27±0.05,P<0.001];The apoptosis rate,the expression of caspase-3 and Bax proteins related to apoptosis,and PP2A expression were significantly increased[(14.02±2.29)% vs.(38.21±3.20)% ],0.81±0.11 vs.1.78±0.24,0.83±0.12 vs.1.72±0.24,0.27±0.02 vs.0.95±0.16,P<0.001].Compared with the miR-383-5p mimic group,BIU-87 cells in the miR-383-5p mimic+pc-CIP2A group significantly increased the cell viability,invasion,migration abilities,the expression of proteins related to invasion,and the expression of Bcl-2 protein[(32.15±2.65)% vs.(50.18±3.77)% ,(82.35±7.01)% vs.(116.30±13.70),(46.12±2.54)% vs.(58.43±3.15)% ,0.22±0.04 vs.0.60±0.08,0.21±0.04 vs.0.5 8±0.06,0.27±0.05 vs.0.64±0.08,P<0.05];The apoptosis rate,the expression of caspase-3,Bax,and PP2A was signifi-cantly reduced in the miR-383-5p mimic+pc-CIP2A group[(38.21±3.20)% (23.15±2.74)% ,1.78±0.24 vs.1.25±0.21,1.72±0.24 vs.1.23±0.18,0.95±0.16 vs.0.60±0.13,P<0.05].The results of dual luciferase experiments showed a corresponding tar-geting relationship between miR-383-5p and CIP2A.Conclusion Increasing the expression of miR-383-5p can inhibit the prolif-eration,invasion and migration of bladder cancer BIU-87 cells,and enhance the ability of apoptosis,which may be achieved by targe-ted regulation of CIP2A.

OBJECTIVE: To analyze the risk factors of in-hospital death in patients with sepsis in the intensive care unit (ICU) based on machine learning, and to construct a predictive model, and to explore the predictive value of the predictive model. METHODS: The clinical data of patients with sepsis who were hospitalized in the ICU of the Affiliated Hospital of Jining Medical University from April 2015 to April 2021 were retrospectively analyzed,including demographic information, vital signs, complications, laboratory examination indicators, diagnosis, treatment, etc. Patients were divided into death group and survival group according to whether in-hospital death occurred. The cases in the dataset (70%) were randomly selected as the training set for building the model, and the remaining 30% of the cases were used as the validation set. Based on seven machine learning models including logistic regression (LR), K-nearest neighbor (KNN), support vector machine (SVM), decision tree (DT), random forest (RF), extreme gradient boosting (XGBoost) and artificial neural network (ANN), a prediction model for in-hospital mortality of sepsis patients was constructed. The receiver operator characteristic curve (ROC curve), calibration curve and decision curve analysis (DCA) were used to evaluate the predictive performance of the seven models from the aspects of identification, calibration and clinical application, respectively. In addition, the predictive model based on machine learning was compared with the sequential organ failure assessment (SOFA) and acute physiology and chronic health evaluation II (APACHE II) models. RESULTS: A total of 741 patients with sepsis were included, of which 390 were discharged after improvement, 351 died in hospital, and the in-hospital mortality was 47.4%. There were significant differences in gender, age, APACHE II score, SOFA score, Glasgow coma score (GCS), heart rate, oxygen index (PaO₂/FiO₂), mechanical ventilation ratio, mechanical ventilation time, proportion of norepinephrine (NE) used, maximum NE, lactic acid (Lac), activated partial thromboplastin time (APTT), albumin (ALB), serum creatinine (SCr), blood urea nitrogen (BUN), blood uric acid (BUA), pH value, base excess (BE), and K⁺ between the death group and the survival group. ROC curve analysis showed that the area under the curve (AUC) of RF, XGBoost, LR, ANN, DT, SVM, KNN models, SOFA score, and APACHE II score for predicting in-hospital mortality of sepsis patients were 0.871, 0.846, 0.751, 0.747, 0.677, 0.657, 0.555, 0.749 and 0.760, respectively. Among all the models, the RF model had the highest precision (0.750), accuracy (0.785), recall (0.773), and F1 score (0.761), and best discrimination. The calibration curve showed that the RF model performed best among the seven machine learning models. DCA curve showed that the RF model exhibited greater net benefit as well as threshold probability compared to other models, indicating that the RF model was the best model with good clinical utility. CONCLUSIONS The machine learning model can be used as a reliable tool for predicting in-hospital mortality in sepsis patients. RF models has the best predictive performance, which is helpful for clinicians to identify high-risk patients and implement early intervention to reduce mortality.
Humans ; Hospital Mortality ; Retrospective Studies ; ROC Curve ; Prognosis ; Sepsis/diagnosis* ; Intensive Care Units

Objective:To analyze the risk factors of acute kidney injury (AKI) in patients with septic shock in intensive care unit (ICU), construct a predictive model, and explore the predictive value of the predictive model.Methods:The clinical data of patients with septic shock who were hospitalized in the ICU of the Affiliated Hospital of Jining Medical College from April 2015 to June 2019 were retrospectively analyzed. According to whether the patients had AKI within 7 days of admission to the ICU, they were divided into AKI group and non-AKI group. 70% of the cases were randomly selected as the training set for building the model, and the remaining 30% of the cases were used as the validation set. XGBoost model was used to integrate relevant parameters to predict the risk of AKI in patients with septic shock. The predictive ability was assessed through receiver operator characteristic curve (ROC curve), and was correlated with acute physiology and chronic health evaluationⅡ(APACHEⅡ), sequential organ failure assessment (SOFA), procalcitonin (PCT) and other comparative verification models to verify the predictive value.Results:A total of 303 patients with septic shock were enrolled, including 153 patients with AKI and 150 patients without AKI. The incidence of AKI was 50.50%. Compared with the non-AKI group, the AKI group had higher APACHEⅡscore, SOFA score and blood lactate (Lac), higher dose of norepinephrine (NE), higher proportion of mechanical ventilation, and tachycardiac. In the XGBoost prediction model of AKI risk in septic shock patients, the top 10 features were serum creatinine (SCr) level at ICU admission, NE use, drinking history, albumin, serum sodium, C-reactive protein (CRP), Lac, body mass index (BMI), platelet count (PLT), and blood urea nitrogen (BUN) levels. Area under the ROC curve (AUC) of the XGBoost model for predicting the risk of AKI in patients with septic shock was 0.816, with a sensitivity of 73.3%, a specificity of 71.7%, and an accuracy of 72.5%. Compared with the APACHEⅡscore, SOFA score and PCT, the performance of the model improved significantly. The calibration curve of the model showed that the goodness of fit of the XGBoost model was higher than the other scores (the calibration curve had the lowest score, with a score of 0.205).Conclusion:Compared with the commonly used clinical scores, the XGBoost model can more accurately predict the risk of AKI in patients with septic shock, which helps to make appropriate diagnosis, treatment and follow-up strategies while predicting the prognosis of patients.

Objective:To investigate the standardization of implementing enteral nutrition in patients with stroke among nurses in Guangdong province.Methods:The results of 175 valid questionnaires submitted by 175 nurses from 60 hospitals in Guangdong province from June to July 2022 were analyzed. Contents of the questionnaires included status of enteral nutrition management of stroke patients (facility, staffing, risk screening, and formulation, implementation and quality control of the plans), nurses' knowledge of enteral nutrition monitoring and complication prevention, follow-up status of enteral nutrition in stroke patients, and nurses' willingness in participating in special training of enteral nutrition.Results:(1) In terms of enteral nutrition facilities and staffing, only 76% and 36% used enteral infusion pumps and heating infusion devices, and only 3% had multidisciplinary team configuration; in terms of risk screening, only 63% had all-patient-screen for enteral nutrition risk; in terms of plan formulation, the proportion of no enteral nutrition process reached to 44%; in terms of plan implementation, the implementation rate of enteral nutrition≤48 h and gastrointestinal function evaluation using gastrointestinal function evaluation scale was less than 50%; in terms of plan quality control, less than 50% Departments had sensitive indexes of enteral nutrition nursing. (2) Among 14 risk management problems of enteral nutrition monitoring and complication prevention, only 3 had accuracy rate higher than 70%. (3) 78% nurses mentioned that patients had experienced enteral nutrition complications, and 77% nurses mentioned that patients/family members had consulted about enteral nutrition related issues; 81% nurses wanted to participate in training or refresher courses related to enteral nutrition, and 88% nurses wanted to join the enteral nutrition Nursing Alliance for stroke patients to serve patients after referral and discharge.Conclusion:The current situation of enteral nutrition management of stroke patients in Guangdong province is not optimistic; nurses lack knowledge of enteral nutrition monitoring and complication prevention, but are strongly willing to participate in the special training of enteral nutrition; relevant measures to further improve the standardized management of enteral nutrition of stroke patients is suggested.

Vaccine hesitation refers to the refusal or delay of vaccination when vaccination services are available and is one of the top ten threats to global health. Vaccine hesitation varies in different countries，and its influencing factors are very complex.However，domestic research on this phenomenon is still in its infancy.This article reviews the phenomenon of vaccine hesitation，the occurrence and influencing factors of vaccine hesitation in some developed countries since the 21st century，to provide the basis for taking effective measures for the phenomenon of vaccine hesitation in China.