Objective To establish primary and simian virus 40(SV40)T antigen transformed human vascular en-dothelial cell models,and provide available resources for endothelial research.Methods Human umbilical vein endothelial cells(HUVEC),human umbilical artery endothelial cells(HUAEC),great saphenous vein endothelial cells(GSVEC)and endothelial cells form endometrium and liver tissue were isolated and cultured respectively.Then,the primary endothelial cells were transformed by lentivirus containing SV40 big T and small T antigens,and continuously subcultured in vitro.The expression of CD31 was detected by flow cytometry,species identification-and mycoplasma detection by PCR,and cell identity was identified by STR detection.The transformed ECs were checked for HLA types.Some of them were tested for RNA expression profile and infected by Cas9 lentivirus to es-tablish stable clones.Results Totally 187 cell lines of transformed HUVEC,1 of transformed HUAEC,5 of trans-formed GSVEC,1 of transformed endothelial cells from endometrium and 1 of transformed endothelial cells from liv-er tissue,and 9 monoclonal HUVEC cell lines stably expressing Cas9 protein were established.All the transformed umbilical endothelial cells were CD31 positive ranging from 20%-90%for 20 cases,while for the rest 168 cases the positive rate was more than 90%.RNA expression revealed stable activation of cell proliferation(cell cycle and DNA synthesis).Their species were identified as human origin.The STR results were consistent with those of the primary culture and unique,and there was no mycoplasma contamination.All these cells could be obtained with the sharing services of National Science and Technology Infrastructure,the National Biomedical Cell-line Resource cen-ters(NSTI-BMCR).Conclusions A series of primary and SV40 T antigen transformed human vascular endothelial cell models have been established,which provide a tool for the study of cardiovascular diseases,inflammation,tumors and immune-related diseases.

Objective:To investigate the diagnostic performance of CT-derived fractional flow reserve (CT-FFR) derived from standard images (STD), images processed by first-generation (SSF1) and second-generation (SSF2) whole-heart motion correction algorithm, respectively.Methods:Patients who underwent both coronary CT angiography (CCTA) and invasive coronary angiography (ICA) with FFR examination within 3 months in Shanghai General Hospital, Shanghai Jiao Tong Univerisity School of Medicine from January 2020 to December 2022 were screened in this retrospective study. Totally of 121 patients (134 lesions) were finally included in the study. CCTA images were reconstructed using iterative reconstruction, iterative reconstruction plus SSF1 and SSF2 algorithms. All images were divided into three groups: STD group, SSF1 group, and SSF2 group. The image quality of the CCTA images was assessed using the Likert scale, and differences between the two groups were compared using the Mann-Whitney U and Kruskal-Wallis test. The correlation and consistency between CT-FFR and FFR were evaluated using Spearman correlation coefficient and Bland-Altman plots. The diagnostic performance of CCTA and CT-FFR from three groups was compared by receiver operating characteristic (ROC) curves. The area under the curve (AUC) was compared using the DeLong test. Results:Compared to the STD group and SSF1 group, the SSF2 group showed the best performance in image quality score (median=3.7). Best correlation ( r=0.652, P<0.001) and consistency (mean difference=0.03) between CT-FFR and FFR were observed in SSF2 group. ROC analysis results revealed that, at the per-lesion level, in the diagnosis of ischemic lesions, the diagnostic performance of CT-FFR in the SSF2 group was significantly better than that of the SSF1 group (AUC=0.88 vs. 0.76, P=0.003), while no significant difference was observed between STD group and SSF1 group ( P=0.125). At the per-patient level, the SSF2 group also demonstrated the highest diagnostic performance. Conclusion:The SSF2 algorithm significantly improved CCTA image quality and enhanced its diagnostic performance for evaluating stenosis severity and CT-FFR calculations.

Objective: To investigate the incidence of microvascular myocardial ischemia in diabetic patients without obstructive coronary artery disease (CAD) and its relationship with angina. Materials and Methods: Diabetic patients and an intermediate-to-high pretest probability of CAD were prospectively enrolled. Non-diabetic patients but with an intermediate-to-high pretest probability of CAD were retrospectively included as controls. The patients underwent dynamic computed tomography-myocardial perfusion imaging (CT-MPI) and coronary computed tomography angiography (CCTA) to quantify coronary stenosis, myocardial blood flow (MBF), and extracellular volume (ECV). The proportion of patients with microvascular myocardial ischemia, defined as any myocardial segment with a mean MBF ≤ of 100 mL/min/100 mL, in patients without obstructive CAD (Coronary Artery Disease–Reporting and Data System [CAD-RADS] grade 0–2 on CCTA) was determined. Various quantitative parameters of the patients with and without diabetes without obstructive CAD were compared. Multivariable analysis was used to determine the association between microvascular myocardial ischemia and angina symptoms in diabetic patients without obstructive CAD. Results: One hundred and fifty-two diabetic patients (mean age: 59.7 ± 10.7; 77 males) and 266 non-diabetic patients (62.0 ± 12.3; 167 males) were enrolled; CCTA revealed 113 and 155 patients without obstructive CAD, respectively. For patients without obstructive CAD, the mean global MBF was significantly lower for those with diabetes than for those without (152.8 mL/min/100 mL vs. 170.4 mL/min/100 mL, P < 0.001). The mean ECV was significantly higher for diabetic patients (27.2% vs. 25.8%, P = 0.009). Among the patients without obstructive CAD, the incidence of microvascular myocardial ischemia (36.3% [41/113] vs. 10.3% [16/155], P < 0.001) and interstitial fibrosis (69.9% [79/113] vs. 33.3% [8/24], P = 0.001) were significantly higher in diabetic patients than in the controls. The presence of microvascular myocardial ischemia was independently associated with angina symptoms (adjusted odds ratio = 3.439, P = 0.037) in diabetic patients but without obstructive CAD. Conclusion Dynamic CT-MPI + CCTA revealed a high incidence of microvascular myocardial ischemia in diabetic patients without obstructive CAD. Microvascular myocardial ischemia is strongly associated with angina.

Objective: The present study aimed to investigate the association between myocardial blood flow (MBF) quantified by dynamic CT myocardial perfusion imaging (CT-MPI) and the increments in heart rate (HR) after stress in patients without obstructive coronary artery disease. Materials and Methods: We retrospectively included 204 subjects who underwent both dynamic CT-MPI and coronary CT angiography (CCTA). Patients with more than minimal coronary stenosis (diameter ≥ 25%), history of myocardial infarction/ revascularization, cardiomyopathy, and microvascular dysfunction were excluded. Global MBF at stress was measured using hybrid deconvolution and maximum slope model. Furthermore, the HR increments after stress were recorded. Results: The median radiation dose of dynamic CT-MPI plus CCTA was 5.5 (4.5–6.8) mSv. The median global MBF of all subjects was 156.4 (139.8–180.4) mL/100 mL/min. In subjects with HR increment between 10 to 19 beats per minute (bpm), the global MBF was significantly lower than that of subjects with increment between 20 to 29 bpm (153.3 mL/100 mL/min vs. 171.3 mL/100 mL/min, p = 0.027). This difference became insignificant when the HR increment further increased to ≥ 30 bpm. Conclusion The global MBF value was associated with the extent of increase in HR after stress. Significantly higher global MBF was seen in subjects with HR increment of ≥ 20 bpm.

Objective: To investigate the diagnostic performance of CT fractional flow reserve (CT-FFR) for myocardial bridging-related ischemia using dynamic CT myocardial perfusion imaging (CT-MPI) as a reference standard. Materials and Methods: Dynamic CT-MPI and coronary CT angiography (CCTA) data obtained from 498 symptomatic patients were retrospectively reviewed. Seventy-five patients (mean age ± standard deviation, 62.7 ± 13.2 years; 48 males) who showed myocardial bridging in the left anterior descending artery without concomitant obstructive stenosis on the imaging were included. The change in CT-FFR across myocardial bridging (ΔCT-FFR, defined as the difference in CT-FFR values between the proximal and distal ends of the myocardial bridging) in different cardiac phases, as well as other anatomical parameters, were measured to evaluate their performance for diagnosing myocardial bridging-related myocardial ischemia using dynamic CT-MPI as the reference standard (myocardial blood flow < 100 mL/100 mL/min or myocardial blood flow ratio ≤ 0.8). Results: ΔCT-FFRsystolic (ΔCT-FFR calculated in the best systolic phase) was higher in patients with vs. without myocardial bridging-related myocardial ischemia (median [interquartile range], 0.12 [0.08–0.17] vs. 0.04 [0.01–0.07], p < 0.001), while CT-FFR systolic (CT-FFR distal to the myocardial bridging calculated in the best systolic phase) was lower (0.85 [0.81–0.89] vs. 0.91 [0.88–0.96], p = 0.043). In contrast, ΔCT-FFRdiastolic (ΔCT-FFR calculated in the best diastolic phase) and CT-FFR diastolic (CT-FFR distal to the myocardial bridging calculated in the best diastolic phase) did not differ significantly. Receiver operating characteristic curve analysis showed that ΔCT-FFRBsystolic had largest area under the curve (0.822; 95% confidence interval, 0.717–0.901) for identifying myocardial bridging-related ischemia. ΔCT-FFRsystolic had the highest sensitivity (91.7%) and negative predictive value (NPV) (97.8%). ΔCT-FFRdiastolic had the highest specificity (85.7%) for diagnosing myocardial bridging-related ischemia. The positive predictive values of all CT-related parameters were low. Conclusion ΔCT-FFR systolic reliably excluded myocardial bridging-related ischemia with high sensitivity and NPV. Myocardial bridging showing positive CT-FFR results requires further evaluation.

Objective To explore a more minimally invasive and economical treatment for atrial septal defect (ASD) through comparing the efficacy and safety in percutaneous transcatheter closure of atrial septal defect procedure,guided by transthoracic echocardiography(TTE) and transesophageal echocardiography(TEE).Methods From March 2014 to September 2017,197 patients with ASD who were feasible to treated with percutaneous transcatheter closure procedure evaluated by preoperative ~ were recruited.TTE showed ASD belonged to secundum(central type) with a maximal diameter range from 2 mm to 35mm,including 82 cases of the ASD without aortic rim.There were 106 patients(47males and 59 females including 37 ASD without aortic rim) with a mean age of 14.2 years(6 months-59 years) old and a mean body weight of 29.5(8.5-64.0)kg were performed percutaneous transcatheter closure of ASD guided by TEE under general anesthesia and endotracheal intubation,and 91 patients(41males and 50 females with 45 ASD without aortic rim inside) with a mean age of 13.8 years(9 months-65 years) old and a mean body weight of 30.4 (9.5-61.0)kg were treated with the percutaneous transcatheter procedure guided by TTE when patients in waking state of local anesthesia(general anesthesia were adopted in patients under 12 years old without intubation).The size of the occluder was selected on the basis of the maximal diameter plus 2-6mm.All 197 cases intraoperation and postoperation data were collected,including complications 、operation time 、operation room stay time and total cost.Results 1 patients in 106 cases of the the TEE group were transferred to small incision on the chest performing transthoracic transcatheter closure of ASD because the difficult stuck of the occluder.3 patients in TEE group transferred to repair under cardiopulmonary bypass(CPB) via small incision on the chest because the difficult stuck of the occluder even in using transthoracic transcatheter closure way.86 patiens in TTE group successfully treated with percutaneous transcatheter closure,and there were 5 failed cases including 2 patients who transferred to TEE guided because of the poor imaging of TTE,another 2 cases treated with transthoracic transcatheter closure of ASD because the difficult stuck of the occluder,and 1 patient performed ASD repair procedure under CPB via small incision on the chest because of the huge ASD without aortic rim and difficult stuck of the occluder.All the 197 patients were cured and discharge from hospital,and there were no complications.There was no significant difference in age,weight,and maximum diameter of ASD between TEE group and TTE group (P ＞ 0.05).Follow-ups were conducted by TTE at month 3 post-operation,and all 197 cases performed no residual shunt of ASD,there were no difference between 2 group.The stay time in the operation room was(68.2 ± 17.3) min in group TEE and (34.7 ± 16.8) min in group TTE,there was obviously shortened the stay time in operation room(P ＜0.01).The total cost of the TTE group was(24.2 ± 2.1) thousand yuan,and the group TEE was(29.3 ± 1.4) thousand yuan,and the cost was significantly reduced in TTE group (P ＜ 0.01).Conclusion The treatment of percutaneous transcatheter closure of ASD guided by TTE is effective and safe,and the feature of more non-invasive and socioeconomic benefits show a broad application prospect.

Zebrafish, a new type of model animal , has been widely used in many fields of biological research be-cause of its low cost , ability of external fertilization , high fecundity , allowance of embryo transplant , and ectogenesis .Re-cently, zebrafish and its embryos have been widely used in ecotoxicological studies and environmental monitoring .Further-more, with the maturation of zebrafish transgenic techniques , a new era has come for environmental pollution monitoring .

OBJECTIVETo observe the effects of Porphyromonas gingivalis (P. gingivalis) ATCC 33277 infection on expression of intercellular adhesion molecule-1 (ICAM-1) in rat vascular smooth muscle cells(VSMC).

METHODSAn infection model of rat VSMC invaded by P. gingivalis was established in vitro. The mRNA of ICAM-1 was measured through reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSCompared with the control group, an apparent and statistically significant increase in expression of ICAM-1 mRNA was observed after 8, 16, and 24 h in P. gingivals-infected rat VSMC (P<0.05). The expression reached its peak at 16 h. Statistically significant differences were observed in the 8 h group and in the other two experimental groups (P<0.05).

CONCLUSIONInfection of P. gingivals in rat VSMC can cause increased expression of ICAM-1, which may have an important function in the progression of atherosclerosis.

Objective To observe the effects of Porphyromonas gingivalis (P. gingivalis) ATCC 33277 infection on ex-pression of intercellular adhesion molecule-1 (ICAM-1) in rat vascular smooth muscle cells(VSMC). Methods An infection model of rat VSMC invaded by P. gingivalis was established in vitro. The mRNA of ICAM-1 was measured through reverse transcription-polymerase chain reaction (RT-PCR). Results Compared with the control group, an apparent and statistically significant increase in expression of ICAM-1 mRNA was observed after 8, 16, and 24 h in P. gingivals-infected rat VSMC (P<0.05). The expression reached its peak at 16 h. Statistically significant differences were observed in the 8 h group and in the other two experimental groups (P<0.05). Conclusion Infection of P. gingivals in rat VSMC can cause increased expres-sion of ICAM-1, which may have an important function in the progression of atherosclerosis.