Objective:To investigate the effects of ghrelin on oxidative stress and ferroptosis in retinal microvascular endothelial cells (HRMEC) under high glucose conditions.Methods:HRMEC were divided into control group, high glucose group, high glucose+ ghrelin group and cultured with conventional medium, 30 mmol/L D-glucose medium, and 30 mmol/L D-glucose+ 10 nmol/L ghrelin medium in vitro for 24 hours accordingly.The cell proliferation was identified by cell counting kit-8 assay.The reactive oxygen species (ROS) levels were detected by flow cytometry.The oxidative stress indexes glutathione (GSH) concentration, malondialdehyde (MDA) superoxide dismutase (SOD) activity and Fe 2+ concentration were detected by the corresponding kit.The mitochondrial structure was observed by transmission electron microscopy.The expression levels of glutathione peroxidase 4 (GPX4) and recombinant solute carrier family 7 member 11 (SLC7A11) proteins were detected by Western blot. Results:The cell proliferation rates of control group, high glucose group and high glucose+ ghrelin group were (100.62±3.40)%, (63.74±4.25)% and (88.19±4.65)%, respectively.The ROS fluorescence intensity of control group, high glucose group and high glucose+ ghrelin group was 15 512.20±1 347.53, 46 457.00±1 072.65 and 22 220.87±1 669.20, GSH concentration was (68.52±7.61), (21.45±1.57) and (55.68±5.15)μmol/L, MDA concentration was (0.79±0.10), (2.47±0.27) and (1.08±0.15)μmol/L, SOD activity was (111.67±10.32), (37.75±5.92) and (97.45±9.12)U/ml, Fe 2+ concentration was (3.02±0.30), (9.45±0.71) and (4.63±0.32)mmol/mgprot, respectively.There were statistically significant differences in cell proliferation rate, ROS fluorescence intensity, GSH, MDA, and Fe 2+ concentrations and SOD activity among the three groups ( F=61.82, 414.59, 61.28, 67.24, 61.64, 146.14; all at P<0.001).Compared with the control group, the cell proliferation rate, GSH concentration and SOD activity were reduced, ROS fluorescence intensity, MDA and Fe 2+ concentrations were increased in the high glucose group, with statistically significant differences (all at P<0.05).Compared with the high glucose group, the cell proliferation rate, GSH concentration and SOD activity were significantly increased, ROS fluorescence intensity, MDA and Fe 2+ concentrations were decreased in the high glucose+ ghrelin group, with statistically significant differences (all at P<0.05).Compared with the control group, the ferroptosis of mitochondria in high glucose group was obvious.Compared with the high glucose group, the mitochondrial status of the high glucose+ ghrelin group was significantly improved.There were significantly differences in the relative expression levels of GPX4 and SLC7A11 proteins in cells among the three groups ( F=63.94, 182.84; both at P<0.001).Compared with the control group, the relative expression levels of GPX4 and SLC7A11 proteins were significantly decreased in the high glucose group and high glucose+ ghrelin group (all at P<0.05).Compared with the high glucose group, the relative expression levels of the two proteins in the high glucose+ ghrelin group were increased, with statistically significant differences (both at P<0.01). Conclusions:Ghrelin can promote proliferation of HRMEC under high glucose conditions, and inhibit high glucose-induced oxidative stress and ferroptosis.

Objective: To explore the clinical and molecular genetic features of a Chinese patient with catecholaminergic polymorphic ventricular tachycardia (CPVT). Methods: Clinical data including resting electrocardiography, echocardiography and treadmill exercise testing of a patient with CPVT admitted to our department in March 2013 were analyzed, and the peripheral venous blood samples of the patient and his family members and 400 ethnicity-matched healthy controls were obtained. All exons and exon-intron boundaries of the six CPVT-related genes including RYR2, CASQ2, TRDN, CALM1, KCNJ2 and ANKB were sequenced to detect the variants related to CPVT. The relationship between the genotypes and phenotypes was analyzed to direct the target therapy. Results: Recurrent syncope induced either by exercise or extreme frightened fear was observed in this patient. There was no positive family history of syncope or sudden death. The resting electrocardiography and echocardiography of the patient were normal, while the exercise testing revealed bidirectional and polymorphic ventricular tachycardia. A cardiac ryanodine receptor gene mutation (R2401H) was identified in this patient, while this mutation was absent in his parents and sister and 400 controls. No variant was detected in the remaining five candidate genes. Treatment with high dose of metoprolol succinate (118.75 mg/d) was effective and patient was free of syncopal attack during the 2 years follow-up. Conclusion This is the first report on RyR2-R2401H mutation in Chinese patient with CPVT, and high dose of metoptolol is the effective therapy option for CPVT related to RyR2 mutation.

Objective: To analyze the clinical features in patients with hypertrophic cardiomyopathy (HCM) combining left ventricular apical aneurysm (LVAA) . Methods: A total of 1194 HCM patients treated in our hospital from 2007-01 to 2013-01 were studied. There were 23 (1.93%) patients combining with LVAA including 19 male and 4 female; coronary artery disease (CAD) was excluded and the patients received echocardiography and coronary angiography (CAG) examinations. Results: There were 21/23 LVAA patients having left mid-ventricular obstruction and 7 of them combining simultaneous left ventricular outflow obstruction.The average pressure gradient in those 21 patients was (56.8 ± 12.9) mmHg and the rest 2 patients suffered from apical hypertrophic cardiomyopathy.The mean maximum thickness of left ventricular wall was (21.8 ± 6.3) mm and the dimension of left ventricle was (39.4 ± 5.2) mm. Electrocardiography showed that 3 patients had paroxysmal ventricular tachycardia;CAG indicated that 6 patients combined with coronary artery muscular bridge at left anterior descending (LAD) artery. The patients were followed-up for (2.7 ± 1.3) years and adverse cardiovascular events occurred in 5 patients during that period. Conclusion: HCM combining LVAA was most frequently happened in patients with left mid-ventricular hypertrophic cardiomyopathy, some of them combining simultaneous left ventricular outflow obstruction and had the higher occurrence rate of adverse cardiovascular events. Early and accurate diagnosis is very important for guiding clinical treatment.

Objective To investigate the expression of BRAF-activated long non-coding RNA (BANCR) in colorectal cancer,and the influence of BANCR on the biological function of HCT116 cells.Methods Fifty-six samples of colorectal cancer specimen (including the cancer tissues and precancerous tissues) were obtained at the First Affiliated Hospital of Nanjing Medical University from March 2012 to June 2013.The expressions of BANCR in all the specimens were detected by qRT-PCR (28 cases in the BANCR-high expression group and 28 cases in the BANCR-low expression group).The relationship between the expressions of BANCR and the clinicopathological factors of colorectal cancer was analyzed.The HCT116 cells were divided into 4 groups after interfering BANCR with lentiviral-mediated shRNA-1 and shRNA-2:interference group 1 (HCT116 cells transfected with LV-shRNA-1),interference group 2 (HCT116 cells transfected with LV-shRNA-2),negative control group (HCT116 cells transfected with lentivirus vector with nonsense sequence) and blank control group (HCT116 cells cultured in RPMI 1640 medium).The proliferation,apoptosis and migration of HCT116 cells in the 4 groups were detected by CCK-8 assay,flow cytometry and Transwell assay,respectively.The comparison between the 2 groups was analyzed by u test,and multiple groups were compared by one-way analysis of variance,repeated measurement analysis of variance and LSD-t test.Multivariate analysis was done by Logistic regression model.The difference between categorical data was compared by chi-square test.Results The relative expression of BANCR in the cancer tissues was 1.6 ± 0.4,which was significantly higher than 0.9 ± 0.7 of the precancerous tissues (u =1 020.000,P ＜ 0.05).The result of univariate analysis showed that the high expression of BANCR was correlated with the lymph node metastasis and tumor stage (x2 =4.595,7.487,P ＜ 0.05).The result of multivariate analysis showed that lymph node metastasis and tumor stage (stage Ⅲ-Ⅳ) were the independent risk factors influencing the high expression of BANCR(OR =4.000,5.914,95％ CI:1.230-12.900,1.685-20.760,P ＜ 0.05).The relative expressions of BANCR of the interference group 1,interference group 2,negative control group and the blank control group were 0.25 ±0.04,0.20±0.06,0.96 ±0.04,0.98 ±0.03,with significant difference among the 4 groups (F =271.610,P ＜ 0.05).The cell proliferation rates at day 6 of the interference group 1,interference group 2 and the negative control group were 80.6％ ± 7.6％,81.2％ ± 5.1％ and 87.9％ ± 13.6％,with no significant difference among the 3 groups (F =0.559,P ＞ 0.05).The apoptotic rates of the interference group 1,interference group 2,negative control group and the blank control group were 4.7％ ± 1.7％,5.1％ ± 1.1％,3.1％ ± 0.6％ and 2.8％ ± 0.9％,with no significant difference among the 4 groups (F =2.881,P ＞ 0.05).The numbers of transmembrane cells of the interference group 1,interference group 2,negative control group and the blank control group were 135 ± 29,107 ± 18,240 ± 24 and 245 ± 22,with significant difference among the 4 groups (F =45.194,P ＜ 0.05).Conclusions BANCR was overexpressed in the HCT116 cells,and the BANCR overexpression was correlated with lymph node metastasis and tumor stage.BANCR can promote the migration of HCT116 cells.BANCR could be an important biomarker for the diagnosis and prognosis of colorectal cancer.