1.Inhibitory effect of miR-146a on high glucose-induced apoptosis of retinal microvascular endothelial cells and its mechanism
Fei GAO ; Zhipeng WU ; Yihua RUAN
Chinese Journal of Experimental Ophthalmology 2021;39(5):398-403
Objective:To investigate the effect of microRNA-146a (miR-146a) on apoptosis of human retinal microvascular endothelial cells (HRMECs) induced by high glucose and its possible molecular mechanism.Methods:HRMECs were cultured in vitro with 5.5 mmol/L D-glucose in the normal control group and 25 mmol/L D-glucose in the high glucose group for 48 hours, respectively.Normally cultured HRMECs were transfected by miR-146a mimics in the high glucose+ miR-146a mimics group or corresponding mimics control in the high glucose+ mimics control group by lipofection and cultured with 25 mmol/L D-glucose for 48 hours, respectively.Real-time fluorescent quantitative polymerase chain reaction (PCR) was performed to detect the expression level of miR-146a.MTT assay and flow cytometry were used to detect the activity and apoptosis of HRMECs, and Western blot was employed to detect the expression levels of apoptosis-associated protein B-cell lymphoma factor-2 (Bcl-2), Bcl-2 related X protein (Bax) and nuclear factor-κB (NF-κB) signaling-related proteins NF-κB p65 and p-NF-κB p65. Results:The relative expression levels of miR-146a were 1.00±0.10, 0.22±0.02, 0.21±0.02 and 0.88±0.09, and the cell viability was (100.00±10.06)%, (68.41±6.67)%, (67.91±6.74)% and (90.46±8.97)%, and the apoptosis rates were (3.11±1.02)%, (27.28±3.56)%, (27.44±4.03)% and (7.29±2.11)% in the normal control group, high glucose group, high glucose+ mimics control group and high glucose+ miR-146a mimics group, respectively.The relative expression levels of miR-146a and the cell viability were significantly lower, and the cell apoptosis rate was significantly higher in the high glucose group than those in the normal control group, with statistical significant differences (all at P<0.05). The relative expression levels of miR-146a and the cell viability were significantly higher, and the cell apoptosis rate was significantly lower in the high glucose+ miR-146a mimics group than those in the high glucose group and the high glucose+ mimics control group, and the differences were statistically significant (all at P<0.05). The relative expression levels of Bax and p-NF-κB p65 protein were significantly higher, the relative expression level of Bcl-2 protein was significantly lower in the high glucose group than those in the normal control group, showing statistically significant differences (all at P<0.05). The relative expression levels of Bax and p-NF-κB p65 protein were significantly lower, and the relative expression level of Bcl-2 protein was significantly higher in the high glucose+ miR-146a mimics group than those in the high glucose group and the high glucose+ mimics control group, and the differences were statistically significant (all at P>0.05). There was no significant difference in the relative expression of NF-κB p65 protein among the groups ( F=0.106, P=0.955). Conclusions:Overexpression of miR-146a may inhibit the apoptosis of HRMECs induced by high glucose, and its mechanism may be related to the inhibition of NF-κB signaling pathway activation.
2.Clinical effect of nebulized acetylcysteine inhalation combined with bronchoscopy in the treatment of elderly patients with severe ventilator-associated pneumonia
Liang WU ; Yihua YU ; Li LI ; Lingyun XU ; Xixi RUAN ; Changbiao LIN ; Jiaping ZHAO
Chinese Journal of Burns 2020;36(4):267-272
Objective:To explore the clinical effect of nebulized acetylcysteine inhalation combined with bronchoscopy in treating elderly patients with severe ventilator-associated pneumonia (VAP).Methods:From January 2016 to December 2017, 80 elderly patients with severe VAP who were hospitalized in Zhejiang Hospital were divided into acetylcysteine+ bronchoscopy group [24 males and 16 females, aged (78±7) years] and bronchoscopy group [26 males and 14 females, aged (80±7) years]using random number table for a prospective cohort study. Patients in bronchoscopy group were treated with bronchoscopy in addition to conventional supportive care of symptoms. Patients in acetylcysteine+ bronchoscopy group received nebulized acetylcysteine inhalation therapy on the basis of the treatment given to patients in bronchoscopy group. Patients in both groups received treatment continuously for 7 days. The simplified clinical pulmonary infection score (CPIS) in both groups was assessed before and after treatment. Venous blood of 10 mL was collected before and after treatment to detect leukocyte count, serum C-reactive protein, and procalcitonin. Arterial blood of 1 mL was collected before and after treatment to detect partial arterial oxygen pressure (PaO 2), partial arterial carbon dioxide pressure (PaCO 2), oxygenation index. The inhalation platform pressure (Pplat), dynamic lung compliance (Cdyn), airway resistance, work of breathing, time of antibiotic use, and time of mechanical ventilation of patients in two groups were recorded before and after treatment. Data were statistically analyzed with chi-square test and t test. Results:(1) The simplified CPIS, leukocyte count, serum C-reactive protein, and procalcitonin of patients in acetylcysteine+ bronchoscopy group were significantly lower than those in bronchoscopy group after treatment ( t=2.32, 2.15, 6.08, 7.12, P<0.05 or P<0.01). The simplified CPIS, leukocyte count, serum C-reactive protein, and procalcitonin of patients in acetylcysteine+ bronchoscopy group and bronchoscopy group after treatment were significantly lower than those before treatment ( t=13.76, 13.60, 12.70, 8.32, 11.44, 14.28, 9.48, 9.50, P<0.01). (2) Compared with bronchoscopy group, patients in acetylcysteine+ bronchoscopy group had significantly higher PaO 2 and oxygenation index ( t=4.14, 2.55, P<0.05 or P<0.01) but significantly lower PaCO 2 ( t=4.36, P<0.01) after treatment. The PaO 2 and oxygenation index of patients in acetylcysteine+ bronchoscopy group after treatment were significantly higher than those before treatment ( t=10.90, 43.72, P<0.01). The PaO 2 and oxygenation index of patients in bronchoscopy group after treatment were also significantly higher than those before treatment ( t=6.55, 43.03, P<0.01). The PaCO 2 of patients in both groups after treatment were significantly lower than those before treatment ( t=21.54, 21.92, P<0.01). (3) The Cdyn of patients in acetylcysteine+ bronchoscopy group after treatment was significantly higher than that in bronchoscopy group ( t=5.41, P<0.01), and Pplat, airway resistance, and work of breathing were significantly lower than those in bronchoscopy group ( t=2.18, 5.46, 2.49, P<0.05 or P<0.01). The Cdyn of patients in both groups after treatment were significantly higher than those before treatment ( t=16.10, 10.90, P<0.01), and Pplat, airway resistance, and work of breathing were significantly lower than those before treatment ( t=21.18, 11.13, 9.32, 15.50, 5.17, 5.97, P<0.01). (4)The time of mechanical ventilation and antibiotic usage of patients in acetylcysteine+ bronchoscopy group were (6.9±1.9)and (8.7±2.8) d, respectively, which were significantly shorter than (10.1±2.2) and (11.6±3.5) d in bronchoscopy group ( t=6.85, 4.09, P<0.01). Conclusions:Nebulized acetylcysteine inhalation combined with bronchoscopy can significantly control the degree of lung infection in elderly patients with severe VAP, improve patients′ respiratory mechanics parameter and blood gas analysis indicator, and shorten the time for mechanical ventilation and antibiotic usage.
3.Predisposing causes and echocardiographic findings of fetal cardiac calcifications
Xiaowei LIU ; Xu YANG ; Xiaoyan HAO ; Yuwei FU ; Yanping RUAN ; Xin WANG ; Xiuqin SONG ; Yihua HE
Chinese Journal of Medical Imaging Technology 2018;34(3):395-398
Objective To discuss the predisposing causes and echocardiographic findings of abnormal cardiac calcifications in fetuses.Methods The echocardiographic and pathological data of 12 fetuses with abnormal cardiac calcifications were retrospectively reviewed,and the pregnancy outcome,predisposing cause and echocardiographic features were analyzed.Results Associated maternal positive anti-SSA/SSB antibody was found in 8 fetuses.The echocardiographic findings of these 8 fetuses were extensive hyper-echogenic thickened atrial wall,interatrial septal and atrioventricular roof.Cardiac myocardium dystrophic calcification was found in 3 fetuses with focal or extensive hyper-echogenic of myocardium.Idiopathic infantile calcification with thickening and hyper-echogenic wall of main pulmonary artery and aorta with obvious stenosis of artery were observed in 1 fetus.One fetus with dystrophic myocardium calcification was continued in pregnancy,1 maternal anti-SSA/SSB antibody (+) baby was born,and 10 fetuses were terminated.Conclusion Maternal anti SSA/ SSB antibody (-) is predominantly associated with fetal cardiac calcification.Fetal abnormal cardiac calcification with different predisposing causes have different echocardiographic features.
4.Application of artificial intelligence in screening the four-chamber view of fetal echocardiography
Xiaoxue ZHOU ; Yingying ZHANG ; Ye ZHANG ; Jiancheng HAN ; Xiaowei LIU ; Xiaoyan GU ; Lin SUN ; Ying ZHAO ; Yanping RUAN ; Yihua HE
Chinese Journal of Ultrasonography 2020;29(8):668-672
Objective:To investigate the value of artificial intelligence in screening normal or abnormal four-chamber view of the fetal heart.Methods:Selecting 3 996 pictures of normal and abnormal end systolic four chamber views and 450 video clips from the database of Beijing Key Laboratory of Fetal Heart Disease Maternal and Fetal Medicine Research in Beijing Anzhen Hospital as training set, test set and verification set to train, test and verify DGACNN model. ①Comparing DGACNN, DGACNN-ALOCC and other classification models(Densenet, Resnet50, InceptionV3, InceptionResnetV2) to detect the model with the most advanced level by recognizing 200 normal pictures and 200 abnormal pictures. ②Fetal echocardiographers were divided into three groups according to their experiences: primary, intermediate and advanced, 3 doctors in each group, and comparing the average score between each group or three groups and DGACNN by recognizing 100 normal pictures and 100 abnormal pictures.Results:①When the the false positive rate(FPR) was in the range of 20%, the recognition accuracy of DGACNN was the highest with 0.850, the recognition accuracy of other models were DGACNN-ALOCC 0.835, Densenet 0.780, Resnet50 0.700, InceptionV3 0.670, InceptionResnetV2 0.650, respectively. ②When FPR was in the range of 20%, the area under ROC curve of DGACNN was the largest with 0.881, the area under ROC curve of other models were DGACNN-ALOCC 0.864, Densenet 0.850, Resnet50 0.822, Inceptionv3 0.779, InceptionResnetV2 0.703, respectively. ③When the FPR was in the range of 20%, the average recognition accuracy of the senior fetal echocardiographer group was the highest with 0.863, followed by DGACNN 0.840, which was higher than the average recognition accuracy of the primary and intermediate groups with 0.760, 0.807; the average recognition accuracy of DGACNN was higher than the total average recognition accuracy of the primary, intermediate and advanced groups with 0.810.Conclusions:Artificial intelligence is accessible in screening four chamber view of fetal echocardiography, with high recognition accuracy.