Purpose To investigate the feasibility of a deep learning framework to automatically analyze echocardiographic color Doppler videos in detecting valvular regurgitation.Materials and Methods This study retrospectively collected echocardiographic images of 1 109 patients with valvular regurgitation in the Fourth Medical Center of PLA General Hospital,from June 2015 to September 2019 as the training and validation sets.A prospective continuous collection of 1 562 echocardiography images was used as the test set in the Fourth Medical Center of PLA General Hospital from May 13 to June 13,2023,including 378 cases of mitral regurgitation and 223 cases of aortic regurgitation.This study developed deep learning networks to establish view classification model and valvular regurgitation recognition model,including the efficiency of section classification of deep learning models.Results The deep learning view classification model in this study could automatically identify two views for diagnosing mitral regurgitation and aortic regurgitation.The recognition accuracy for the parasternal long axis color Doppler view and the apical four chamber mitral color Doppler view was 1.00 and 0.93,respectively.The sensitivity,specificity,accuracy and area under the curve of the deep learning model for diagnosing mitral regurgitation were 0.847,0.852,0.849 and 0.930,respectively.The sensitivity,specificity,accuracy and area under the curve of the deep learning model in diagnosing aortic regurgitation were 0.857,0.861,0.859 and 0.940,respectively.Conclusion Deep learning algorithms can automatically identify valvular regurgitation and have the potential to become a screening tool for valvular heart disease.

Objective:To investigate the effect of tyrosine kinase receptor binding protein B3 (Ephrin-B3) on the expressions of postsynaptic density protein 95 (PSD95), N-methyl-D-aspartic acid (NMDA) receptor subunit 2A and 2B(NR2A/NR2B) and synaptic ultrastructure of hippocampal neurons in the dentate gyrus (DG)area of epileptic rats.Methods:A total of 96 SPF-grade adult male Sprague Dawley(SD) rats were divided into blank group, epilepsy group, lentiviral vectors control group and Ephrin-B3 overexpression group( n=24 in each group) according to the random number table.Another 36 adult male SD rats were randomly divided into recombinant adeno-associated virus control group and Ephrin-B3 suppression group( n=18 in each group). The viruses were stereotaxically injected into bilateral hippocampus DG area of rats and epileptic model was established by intraperitoneal injection of lithium chloride and pilocarpine.Racine standard grading method was used to evaluate seizure rate and the duration and latency of epileptic rats were observed. Immunofluorescence staining was used to observe expression levels of PSD95, NR2A and NR2B protein in hippocampus DG area of rats.PCR and Western blot were used to detect the expression levels of PSD95, NR2A and NR2B mRNA and protein in hippocampus of rats. Transmission electron microscopy was used to observe the ultrastructure of hippocampal neurons in rats. GraphPad Prsim 8 software was used for statistical analysis. Results:(1) The results of Ephrin-B3 overexpression experiment: there were statistically significant differences in the seizure latency and duration among the four groups( F=368.30, 120.00, both P<0.01). The seizure latency in Ephrin-B3 overexpression group was longer than that of the epilepsy group and lentiviral vectors control group((31.32±1.10)d, (24.09±2.02)d, (21.42±0.79)d)(all P<0.01), while the seizure duration in Ephrin-B3 overexpression group was shorter than that of the epilepsy group and lentiviral vectors control group((26.85±1.14)s, (40.40±1.26)s, (36.50±5.50)s)(all P<0.05). Immunofluorescence staining showed the average fluorescence intensity of PSD95, NR2A and NR2B in Ephrin-B3 overexpression group were higher than that in the lentiviral vectors control group(all P<0.01). PCR and Western blot both showed that the Ephrin-B3 overexpression group had higher mRNA expression levels of PSD95, NR2A and NR2B than that in lentiviral vectors control group(all P<0.05). The results of transmission electron microscopy showed the number of synapses in the Ephrin-B3 overexpression group was greater than that in lentiviral vectors control group((9.00±1.00), (6.00±1.00))( P<0.05), synaptic gap was narrower than that in lentiviral vectors control group ((31.60±1.45)nm, (36.43±2.22)nm)(all P<0.05). (2)The results of the Ephrin-B3 suppression experiment: the evaluation of seizures in rats showed that there were statistically significant differences in the seizure latency and duration between recombinant adeno-associated virus control group and Ephrin-B3 suppression groups( t=3.88, 3.93, both P<0.05). The seizure latency in Ephrin-B3 suppression group was shorter than that of recombinant adeno-associated virus control group ((17.10±1.88)d, (23.50±2.15)d)( P<0.05). The seizure duration in Ephrin-B3 suppression group was longer than that of recombinant adeno-associated virus control group((55.16±5.48)s, (42.06±1.83)s)( P<0.05). Western blot results showed that the Ephrin-B3 suppression group had lower protein expression levels of PSD95, NR2A and NR2B than those in recombinant adeno-associated virus control group(all P<0.01). The results of transmission electron microscopy showed the numbers of synapses in the Ephrin-B3 suppression group were less than recombinant adeno-associated virus control group((3.33±0.58), (4.66±0.58))( P<0.05), synaptic gap were narrower than recombinant adeno-associated virus control group((37.27±0.97)nm, (33.33±1.46)nm)( P<0.05). Conclusion:Ephrin-B3 can attenuate seizures and upregulate the synaptic related proteins and glutamate receptor expression, improving synaptic structure.

Objective:To investigate the effect of tyrosine kinase receptor binding protein B3 (Ephrin-B3) on the expressions of postsynaptic density protein 95 (PSD95), N-methyl-D-aspartic acid (NMDA) receptor subunit 2A and 2B(NR2A/NR2B) and synaptic ultrastructure of hippocampal neurons in the dentate gyrus (DG)area of epileptic rats.Methods:A total of 96 SPF-grade adult male Sprague Dawley(SD) rats were divided into blank group, epilepsy group, lentiviral vectors control group and Ephrin-B3 overexpression group( n=24 in each group) according to the random number table.Another 36 adult male SD rats were randomly divided into recombinant adeno-associated virus control group and Ephrin-B3 suppression group( n=18 in each group). The viruses were stereotaxically injected into bilateral hippocampus DG area of rats and epileptic model was established by intraperitoneal injection of lithium chloride and pilocarpine.Racine standard grading method was used to evaluate seizure rate and the duration and latency of epileptic rats were observed. Immunofluorescence staining was used to observe expression levels of PSD95, NR2A and NR2B protein in hippocampus DG area of rats.PCR and Western blot were used to detect the expression levels of PSD95, NR2A and NR2B mRNA and protein in hippocampus of rats. Transmission electron microscopy was used to observe the ultrastructure of hippocampal neurons in rats. GraphPad Prsim 8 software was used for statistical analysis. Results:(1) The results of Ephrin-B3 overexpression experiment: there were statistically significant differences in the seizure latency and duration among the four groups( F=368.30, 120.00, both P<0.01). The seizure latency in Ephrin-B3 overexpression group was longer than that of the epilepsy group and lentiviral vectors control group((31.32±1.10)d, (24.09±2.02)d, (21.42±0.79)d)(all P<0.01), while the seizure duration in Ephrin-B3 overexpression group was shorter than that of the epilepsy group and lentiviral vectors control group((26.85±1.14)s, (40.40±1.26)s, (36.50±5.50)s)(all P<0.05). Immunofluorescence staining showed the average fluorescence intensity of PSD95, NR2A and NR2B in Ephrin-B3 overexpression group were higher than that in the lentiviral vectors control group(all P<0.01). PCR and Western blot both showed that the Ephrin-B3 overexpression group had higher mRNA expression levels of PSD95, NR2A and NR2B than that in lentiviral vectors control group(all P<0.05). The results of transmission electron microscopy showed the number of synapses in the Ephrin-B3 overexpression group was greater than that in lentiviral vectors control group((9.00±1.00), (6.00±1.00))( P<0.05), synaptic gap was narrower than that in lentiviral vectors control group ((31.60±1.45)nm, (36.43±2.22)nm)(all P<0.05). (2)The results of the Ephrin-B3 suppression experiment: the evaluation of seizures in rats showed that there were statistically significant differences in the seizure latency and duration between recombinant adeno-associated virus control group and Ephrin-B3 suppression groups( t=3.88, 3.93, both P<0.05). The seizure latency in Ephrin-B3 suppression group was shorter than that of recombinant adeno-associated virus control group ((17.10±1.88)d, (23.50±2.15)d)( P<0.05). The seizure duration in Ephrin-B3 suppression group was longer than that of recombinant adeno-associated virus control group((55.16±5.48)s, (42.06±1.83)s)( P<0.05). Western blot results showed that the Ephrin-B3 suppression group had lower protein expression levels of PSD95, NR2A and NR2B than those in recombinant adeno-associated virus control group(all P<0.01). The results of transmission electron microscopy showed the numbers of synapses in the Ephrin-B3 suppression group were less than recombinant adeno-associated virus control group((3.33±0.58), (4.66±0.58))( P<0.05), synaptic gap were narrower than recombinant adeno-associated virus control group((37.27±0.97)nm, (33.33±1.46)nm)( P<0.05). Conclusion:Ephrin-B3 can attenuate seizures and upregulate the synaptic related proteins and glutamate receptor expression, improving synaptic structure.

Purpose To investigate the feasibility of a deep learning framework to automatically analyze echocardiographic color Doppler videos in detecting valvular regurgitation.Materials and Methods This study retrospectively collected echocardiographic images of 1 109 patients with valvular regurgitation in the Fourth Medical Center of PLA General Hospital,from June 2015 to September 2019 as the training and validation sets.A prospective continuous collection of 1 562 echocardiography images was used as the test set in the Fourth Medical Center of PLA General Hospital from May 13 to June 13,2023,including 378 cases of mitral regurgitation and 223 cases of aortic regurgitation.This study developed deep learning networks to establish view classification model and valvular regurgitation recognition model,including the efficiency of section classification of deep learning models.Results The deep learning view classification model in this study could automatically identify two views for diagnosing mitral regurgitation and aortic regurgitation.The recognition accuracy for the parasternal long axis color Doppler view and the apical four chamber mitral color Doppler view was 1.00 and 0.93,respectively.The sensitivity,specificity,accuracy and area under the curve of the deep learning model for diagnosing mitral regurgitation were 0.847,0.852,0.849 and 0.930,respectively.The sensitivity,specificity,accuracy and area under the curve of the deep learning model in diagnosing aortic regurgitation were 0.857,0.861,0.859 and 0.940,respectively.Conclusion Deep learning algorithms can automatically identify valvular regurgitation and have the potential to become a screening tool for valvular heart disease.

Objective To investigate the relationship of miRNA gene polymorphisms with blood pressure(BP)responses to the sodium and potassium diet intervention.Methods In 2004,we recruited 514 participants from 124 families in seven villages of Baoji,Shaanxi Province,China.All subjects were given a three-day normal diet,followed by a seven-day low-salt diet,a seven-day high-salt diet,and finally a seven-day high-salt and potassium supplementation.A total of 19 miRNA single nucleotide polymorphisms(SNPs)were selected for analysis.Results Throughout the sodium-potassium dietary intervention,the BP of the subjects fluctuated across all phases,showing a decrease during the low-salt period and an increase during the high-salt period,followed by a reduction in BP subsequent to potassium supplementation during the high-salt diet.MiR-210-3p SNP rs 12364149 was significantly associated with systolic BP(SBP),diastolic BP(DBP)and mean arterial pressure(MAP)responses to low-salt diet.MiR-4638-3p SNP rs6601178 was significantly associated with SBP while miR-26b-3p SNP rs115254818 was significantly associated with MAP responses to low-salt intervention.In addition,miR-26b-3p SNP rs115254818 was significantly correlated with SBP,DBP and MAP responses to high-salt intervention.MiR-1307-5p SNPs rs1 1191676 and rs2292807 were associated with SBP and MAP responses to high-salt diet.MiR-4638-3p SNP rs6601178,miR-210-3p SNP rs12364149,miR-382-5p SNP rs4906032 and rs4143957 were significantly associated with SBP response to high-salt diet.In addition,miR-26b-3p SNP rs115254818 was significantly associated with SBP,DBP and MAP responses to potassium supplementation.MiR-1307-5p SNPs rs11191676,rs2292807,and miR-19a-3p SNP rs4284505 were significantly associated with SBP responses to high-salt and potassium supplementation.Conclusion miRNA gene polymorphisms are associated with BP response to sodium and potassium,suggesting that miRNA genes may be involved in the pathophysiological process of salt sensitivity and potassium sensitivity.

This paper explained the etiology and mechanism of atrial fibrillation based on the theory of "deficiency qi stagnation" for directing the treatment. It is believed that “deficiency qi” is the root of atrial fibrillation， which can be divided into deficiency of pectoral qi in heart and lungs of the upper jiao， deficiency of center qi in spleen and stomach of the middle jiao， and deficiency of original qi in kidney of the lower jiao， and stagnation of stasis， phlegm dampness， cold dampness and other pathogenic qi as the pathological basis of atrial fibrillation， which lead to the development of atrial fibrillation by constraint to heat， or stagnation of cold and dampness， or pathogens stagnation in heart and lung. The therapeutic principles is to supplement deficiency and remove stagnation， and Qingliang Buqi Tiaomai Decoction （清凉补气调脉汤） is often used to supplement the center and boost qi， rectify qi and unblock vessels； Qingliang Huashi Tiaomai Decoction （清凉化湿调脉汤） is often used to strengthen the middle and remove phlegm， dry dampness and unblock vessels； Ziyang Wenhua Tiaomai Decoction （滋养温化调脉汤） is often used to bank up original qi， supplement yin and tonify yang， dissipate cold and unblock vessels.

Myocardial fibrosis is characterized by pathological remodeling of extracellular matrix,which is a common pathological change during the development of various cardiovascular diseases.Qi transformation dysfunction in the zang-fu organs,subtle substance accumulation,and endogenous turbid evil production lead to the occurrence of diseases.The theory of turbid blood is widely used to elucidate the pathological changes of diseases and guide the prevention and treatment.Turbid blood,as a special pathogenic factor and pathological product,plays a crucial role in the oxidative stress process of myocardial fibrosis.Qi deficiency of the heart and spleen,stagnation of turbid blood,impaired blood circulation in the heart,and the inability to maintain the oxidative-reductive system balance of myocardial cells are the root causes of disease onset.Accumulation of turbid blood,intermingled phlegm and blood stasis,blockage of heart vessels,and accumulation of metabolic waste products contribute to disease progression.Prolonged turbidity accumulation leads to cardiac enlargement,scattered mental state,and pathological remodeling of the extracellular matrix,indicating a severe disease stage.Early treatment focuses on strengthening the vital qi and spleen,reducing turbidity and recovering clarity.In the middle stage,the key is to resolve phlegm,eliminate stasis,and promote clarity while removing turbidity.In the late stage,detoxification,turbidity elimination,and restoring clarity are emphasized.By adhering to the characteristics of the pathological mechanism and using traditional Chinese medicine intervention,it is possible to suppress oxidative stress,prevent pathological remodeling of the extracellular matrix,and improve myocardial fibrosis.

Objective To investigate the progress of two-dimensional echocardiographic parameters in patients with different severity of aortic stenosis.Methods A retrospective analysis was per-formed on 96 patients diagnosed with aortic stenosis with at least 2 times of transthoracic echo-cardiography(interval ≥1 year)in Department of Cardiology,Fourth Medical Center of Chinese PLA General Hospital from March 2017 to December 2023.According to aortic stenosis severity,they were divided into a mild group(72 cases),a moderate group(14 cases)and a severe group(10 cases).Peak pressure gradient(PPG)across aortic valve,Vmax,mean aortic valve pressure gradient(ΔPm),pulmonary artery systolic pressure(PASP)were collected,and the changes and annual progress of these echocardiographic parameters at baseline and before and after follow-up were analyzed.Results The values of IVST,LVPWT,Vmax,aortic valve PPG and ΔPm were sig-nificantly increased in the mild,moderate and severe stenosis groups in turn(P＜0.05,P＜0.01).The values of Vmax,PPG and ΔPm were significantly lower in the mild stenosis group than the moderate and severe stenosis groups,and the LVPWT value was obviously lower in the mild ste-nosis group than the severe stenosis group(P＜0.05).The aortic valve PPG and ΔPm values at follow-up were significantly higher than those before the follow-up in the three stenosis groups(P＜0.05,P＜0.01).After follow-up,the Vmax values in mild and moderate stenosis groups were notably higher than before(P＜0.01).The PASP value at follow-up was significantly higher than before in the severe stenosis group(P＜0.05).The annual progression rate of Vmax,PASP,LVEF were gradually increased in the mild,moderate,and severe stenosis groups(P＞0.05).The annual progression rate of ΔPm was gradually increased in the three groups in turn(2.30±1.77 mm Hg/year vs 2.40±1.18 mm Hg/year vs 6.08±1.70 mm Hg/year,P＜0.05).Conclusion As the severity of baseline aortic stenosis increases,obvious changes are observed in cardiac structure and function.Before and after follow-up,the serious the aortic stenosis severity is,the faster the annual progression rates of Vmax,PPG,LVEF and PASP are.

Objective:To investigate the role and mechanism of mitochondrial DNA N6-methyladenine (6mA) in the hippocampal neurons in vascular cognitive impairment.Methods:(1) In vivo experiments: SPF male rats were randomly divided into sham-operated group and chronic cerebral hypoperfusion (CCH) group ( n=12). CCH models in the CCH group were established by ligating bilateral carotid arteries, while rats in the sham-operated group were only bilaterally dissected without ligation. Exploratory ability was detected by open field test 50 d after modeling, cognitive function was evaluated by novel object recognition test 51-53 d after modeling, and learning and memory abilities were tested by Morris water maze 54-59 d after modeling. And then, rats were sacrificed; ATP concentration and reactive oxygen species (ROS) level in the hippocampal tissues were detected, and neuron apoptosis in the hippocampal CA1 area was detected by TUNEL. (2) In vitro experiments: HT-22 cells were divided into normal control (NC) group, oxygen-glucose deprivation (OGD) group, OGD+siControl group, and OGD+siMETTL4 group. Cells in the NC group were cultured routinely, cells in the OGD group were subjected to low sugar and low oxygen for 12 h, and cells in the OGD+siControl group and OGD+siMETTL4 group were, respectively, transfected with NC-siRNA or METTL4-siRNA after being subjected to low sugar and low oxygen for 12 h. Mitochondria morphology was observed by transmission electron microscopy, ROS was detected by flow cytometry, mitochondria membrane potential was detected by JC-1 fluorescent staining, and mitochondrial complex I and III activity was detected by kit. (3) In vivo and in vitro experiments: METTL4 and DNA 6mA expressions in neuronal mitochondria of rat hippocampal tissues and mitochondria of HT-22 cells were detected by immunofluorescent staining and Western blotting. Results:(1) CCH rats had cognitive impairment: compared with the sham-operated group, CCH group had significantly increased frequency of entering the central area and reduced time in exploring new objects in open field experiment,and significantly decreased frequency of crossing the platform and prolonged escape latency in water maze experiment ( P<0.05). Compared with rats in the sham-operated group, rats in the CCH group had significantly decreased hippocampal ATP content ([18.820±1.177] nmol/L vs. [10.190±0.519] nmol/L) and increased ROS content ([4 488.00±255.70] AU vs. [11 644.00±530.20] AU, P<0.05). TUNEL results showed that the number of apoptotic neurons in the hippocampal CA1 area of CCH group was obviously increased than that in sham-operated group. Immunofluorescent staining results showed that 6mA and METTL4 mainly distributed in the mitochondria of hippocampal neurons in CCH group, and the 6mA and METTL4 expressions were obviously increased compared with those in the sham-operated group. Western blotting results showed that METTL4 expression in the hippocampal mitochondria of CCH group was significantly higher than that in the sham-operated group (1.729±0.168 vs. 1.000±0.000). (2) In vitro experiment: under transmission electron microscope, compared with the NC group, HT-22 cells in the OGD group showed obvious mitochondrial ridge disappearance, membrane rupture and vacuolation. Compared with the OGD group, the OGD+siMETTL4 group had significantly increased ATP production, decreased mtROS production, increased mitochondrial membrane potential, and increased mitochondrial complex I and III activities ( P<0.05). Immunofluorescent staining results showed that the mtDNA 6mA and METTL4 expressions in the OGD group were obviously higher than those in the NC group, and both mainly expressed in the mitochondria; mtDNA 6mA expression in the OGD+siMETTL4 group was obviously lower than that in OGD group. Western blotting results showed that METTL4 expression in the OGD+siMETTL4 group was significantly higher than that in the OGD group (1.578±0.261 vs. 2.970±0.280). Conclusion:Specific high expression of methylase METTL4 in hippocampal neurons of rats with cognitive impairment after CCH promotes the increased mtDNA 6mA expression and leads to mitochondrial energy metabolism disorders and increased ROS, which is speculated to be one of the mechanisms causing vascular cognitive impairment.

【Objective】 Corin, a transmembrane serine protease that can cleave atrial natriuretic peptide precursor (pro-ANP) into atrial natriuretic peptide with smaller bioactive molecules, participates in the pathophysiological process of hypertension and cardiac hypertrophy. The purpose of this study was to explore the relationship of Corin gene variation with blood pressure responses to sodium and potassium dietary interventions. 【Methods】 In 2004, we recruited 514 participants from 124 families in 7 villages of Baoji, Shaanxi Province, China. All the subjects received a 3-day normal diet, a 7-day low-salt diet, a 7-day high-salt diet, and finally a 7-day high-salt and potassium supplementation. Fifteen single nucleotide polymorphisms (SNPs) of Corin gene were selected for final analysis. 【Results】 SNPs rs12509275 were significantly associated with diastolic blood pressure (DBP) response to low-salt diet, while rs3749584 was associated with pulse pressure (PP) response to low-salt diet.SNP rs3749584 and rs10517195 were significantly associated with PP response to high-salt diet. In addition,rs17654278 were significantly associated with systolic blood pressure (SBP) response to high-salt and potassium supplementation, rs2271037 was significantly correlated with DBP responses to high-salt and potassium supplementation, and rs4695253, rs12509275, rs2351783, rs36090894 were significantly associated with PP response to high-salt and potassium supplementation. 【Conclusion】 Corin gene polymorphisms were associated with blood pressure response to sodium and potassium, suggesting that Corin gene may be involved in pathophysiological process of salt sensitivity and potassium sensitivity.