Chaofeng Sun

Drug-Induced Long QT Syndrome in Women

Congenital long QT syndromes (LQTS) are inherited heart diseases that can present as palpitations, syncope (fainting), seizures, cardiac arrest, and sudden death. Acquired LQTS mostly occurs as a result of exposure to an environmental stressor that is responsible for the excessive prolongation of the QT interval. The most common environmental stressor is adverse drug reactions, which can lead to drug-induced LQTS (di-LQTS). Female gender has been increasingly recognized as an independent risk factor for di-LQTS, which in females is influenced by other factors, including age, menstrual cycle, and hormone replacement therapy. The estrogen-mediated reduced repolarization reserve in women is believed to be responsible for their higher susceptibility to di-LQTS. More studies, especially randomized trials, should be carried out to confirm these findings, and elucidate the clinical impact of gender disparity in modifying the risk of di-LQTS in women, with the ultimate goal of promoting the clinical safety of medication. In this article, we review current knowledge about di-LQTS, specifically in women, and discuss methods for the prevention of di-LQTS in females.

Early use of granulocyte colony stimulating factor improves survival in a rabbit model of chronic myocardial ischemia

BACKGROUND Granulocyte colony stimulating factor (G-CSF) improves the survival of animals with myocardial infarction by inducing bone marrow stem cell mobilization and homing to infarcted areas. However, its precise mechanisms and direct effects on the ischemic myocardium remain unclear. In this study we investigated the direct effects and mechanisms of G-CSF in a rabbit model of chronic myocardial ischemia. METHODS Myocardial ischemia models were created by partial ligation of the left anterior descending coronary artery in Japanese white male rabbits. Rabbits were subcutaneously injected with 10 μg/kg of G-CSF (G-CSF group) or saline (control group) for 6 days after myocardial ischemia. Direct effects of G-CSF were analyzed by immunohistochemistry and terminal dUTP nick end-labeling (TUNEL). RESULTS Rabbits in the G-CSF group exhibited 75% survival compared to 40% in the control group (p<0.05). Immunohistochemistry of the ischemic myocardium showed increased homing of CD34+ cells on day 7 post-surgery and more vessels on day 28 post-surgery by anti-von Willebrand factor staining in the G-CSF group compared with the control group. Furthermore, an increased percentage of CD34+ cells were observed in peripheral blood and upregulation of vascular endothelial growth factor expression in ischemic tissue in the G-CSF group compared with the control group. TUNEL showed that the apoptotic index in the ischemic myocardium decreased in the G-CSF group compared with the control group on day 28 post-surgery. CONCLUSIONS In addition to increasing stem cell mobilization and homing to ischemic myocardium, G-CSF treatment after myocardial ischemia improves survival by accelerating neovascularization and reducing apoptosis.

CD34+ cell mobilization and upregulation of myocardial cytokines in a rabbit model of myocardial ischemia

BACKGROUND Studies have suggested that myocardial infarction may induce bone marrow stem cell mobilization and homing to the infarcted area, contributing to myocardial repair and tissue regeneration. Despite some encouraging results using stem cell therapy for myocardial regeneration in humans and animals, the mechanisms behind this activity remain unclear. In this study, we investigate stem cell mobilization and homing in ischemic myocardium, and investigate the involvement of cytokines TNFα and VEGF in this process. METHODS AND RESULTS Myocardial ischemia models were created by partial ligation of the left anterior descending coronary artery in Japanese white male rabbits. Immunohistochemistry analysis of ischemic myocardium showed the presence of VEGF and TNFα along with homing of CD34 positive (CD34+) cells to the region in the 7 days following surgery. During the same period, an increase in percentage of CD34+ cells in peripheral blood and in VEGF and TNFα mRNA expression in ischemic tissue was observed in animals that underwent partial LAD ligation. Terminal dUTP nick end-labeling (TUNEL) showed that cell apoptosis in the ischemic myocardium decreased between days 7 and 28 following surgery. None of these changes were observed in animals that underwent sham operation. CONCLUSIONS In the early stages of myocardial ischemia, bone marrow stem cells are mobilized and home to ischemic myocardium with a concomitant increase in expression of cytokines VEGF and TNFα. Furthermore, cell apoptosis occurs in the ischemic myocardium, possibly due to the activity of TNFα which is thought to induce cardiomyocyte apoptosis.

New iPSC for old long QT syndrome modeling: Putting the evidence into perspective

Induced pluripotent stem cells (iPS cells or iPSCs) are typically derived by transfection of certain stem cell-associated genes into non-pluripotent cells, such as adult fibroblasts (typically adult somatic cells). Various diseases can be modeled through iPSC technology. The important implication of iPSCs to offer an unprecedented opportunity to recapitulate pathologic human tissue formation in vitro has generated great excitement and interest in the whole biomedical research community. Long QT syndrome (LQTS), an inherited heart disease, is characterized by prolonged QT interval on a surface electrocardiogram. LQTS presents with life-threatening cardiac arrhythmias, which can lead to fainting, syncope, and sudden death. The iPSC-derived cardiomyocytes from LQTS patients offer a potentially unlimited source of materials for biomedical study. They can be used to recapitulate complex physiological phenotypes, probe toxicological testing and drug screening, clarify the novel mechanistic insights and may also rectify gene defects at the cellular and molecular level. Despite the emerging challenges, iPSC technology has been increasingly recognized as a valuable and growing toolkit for modeling LQTS over other various models of human diseases.

Targeting Amino Acids Metabolic Profile to Identify Novel Metabolic Characteristics in Atrial Fibrillation

Background: Atrial fibrillation (AF) is the most common cardiac arrhythmia whose incidence is on the rise globally. However, the pathophysiologic mechanism of AF remains poorly understood and there has been a lack of circulatory markers to diagnose and predict prognosis of AF. In the present study, by measuring metabolic profile and analyzing plasma amino acid levels in AF patients, we sought to determine whether amino acid metabolism was correlated to the occurrence of AF. Methods: Consecutive patients admitted to hospital for AF were enrolled. Plasma samples were obtained after overnight fast and a profile of 61 amino acids was then measured using gas chromatography/mass spectrometry (GC/MS). Results: Twenty-three AF and thirty-seven control patients were enrolled in the study. A number of plasma amino acids were altered in AF, which showed significant prediction value for AF. Intriguingly, circulating 4-hydroxypyrrolidine-2-carboxylic was gradually lowered with the persistence of AF. Plasma amino acid levels were more strongly correlated with each other in AF as compared with control. Conclusion: By utilizing non-target metabolic profile surveys, we have found a number of altered amino acids, which exhibit diagnostic value for AF. Enhanced amino acids correlation network further identified AF as a metabolism disorder.

Construction of a HERG mutant L539fs/47-*558W pEGFP vector and the expression of the fusion protein in HEK293 cells

Abstract Objective To construct a human ether-a-go-go-related gene (HERG) nonsense mutant L539fs/47-*558W into the autonomously fluorescent, eukaryotic expression vector pEGFP-C2, and to verify expression of the reconstruct in human embryonic kidney-293 (HEK293) cells. Methods The mutational fragment was subcloned into pEGFP-C2-HERG by double digestion of Sbf I, Eco91 I and rejoining of T4 ligase. After verification, the recombinant pEGFP-C2-L539fs/47-*558W and pEGFP-C2-HERG were respectively transfected into HEK293 cells for 48 h by the Lipofect method to observe the expression location of the fusion protein by laser confocal imaging scanning in vivo. pcDNA3 -L539fs/47-*558W and pcDNA3-HERG were transfected to observe the expression location of the HERG protein by immunofluoresceoce. The mutant protein size was determined by Western blotting. Results The about 1 kb-sized mutation region cDNA fragment from pcDNA3-L539fs/47-*558W and the about 7.2 kb-sized target vector fragment from pcDNA3-HERG were ligated after purification and gel recovery. pEGFP-C2-L539fs/47*-558W, approximately 8.2 kb, was demonstrated successfully been constructed under agarose gel electrophoresis and further sequencing. Laser confocal imaging showed that pEGFP-C2-HERG was mainly expressed in the membrane, whereas truncated mutant-type HERG in the pEGFP-C2 vector was partially located in the cytoplasm, the others were transported to the cell membrane in living HEK293 cells. The same as the immunofluoresceoce results after transfection of pcDNA3-HERG and pcDNA3-L539fs/47-558W. Wild-type HERG-GFP fusion protein expressed 160 and 180 kDa bands. The mutant and mutant-GFP fusion proteins were 70 and 100 kDa, respectively. Conclusion pEGFP-C2-L539fs/47-*558W was successfully constructed by double digestion method GFP had no effect on its protein expression and trafficking in HEK293 cells, which laid a foundation for the further study on L539fs/47-*558W.

Induced pluripotent stem cell models of long QT syndrome

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