Comparison of two methods for culturing neonatal rat cardiac fibroblasts

AIM:To establish and compare the methods for culturing neonatal rat cardiac fibroblasts culture. METHODS:Neonatal rat hearts were isolated by collagenase+trypsin or trypsin digestion. The cardiomyocytes and cardiac fibroblasts were isolated by different attachment techniques. The cellular morphology, purity and reactions to the reagent were tested. RESULTS:For morphology and purity, no difference between the 2 methods was observed, though more myocytes were harvested by the method of collagenase+trypsin digestion. For the cellular responses to reagent, the cardiac fibroblasts harvested with trypsin digestion had more potent proliferative ability than those with collagenase+trypsin digestion. CONCLUSION:There is no difference of cellular morphology and purity in the cardiac fibroblasts isolated by collagenase+trypsin digestion and trypsin digestion, but the fibroblasts with trypsin digestion have more potent proliferative ability.     

5年生种植人参对成年大鼠心功能和血流动力学的影响

目的观察5年生种植人参食用对成年大鼠心功能和血流动力学的影响,探讨人参食用对机体心血管系统功能的调节作用。方法正常成年Wistar大鼠随机分为10组,即:对照组,5年生种植人参原粉小、中、大剂量(0.25、0.5、1.0g生药/kg)组,5年生种植人参水提物小、中、大剂量(0.25、0.5、1.0g生药/kg)组,5年生种植人参醇提物小、中、大剂量(0.25、0.5、1.0g生药/kg)组,每组14只。各组大鼠连续灌胃给药3个月,观察心功能和血流动力学变化。结果与对照组比较,人参原粉和人参醇提物可使正常成年大鼠动脉收缩压(SBP)、左室收缩压(LVSP)、左心室内压最大上升和下降速率(±dp/dtmax)均明显增高(P0.05或P0.01),对舒张压(DBP)、平均动脉压(MAP)、左心室舒张末压(LVEDP)及心率(HR)均无明显影响(P0.05)。人参水提物各剂量组对SBP、DBP、MAP、LVSP、±dp/dtmax、LVEDP及HR均无明显影响(P0.05)。结论人参原粉和人参醇提物对正常成年大鼠具有明显的正性肌力(强心)作用,人参水提物对正常成年大鼠的血压及心脏心收缩和舒张功能均无增强或降低作用。     

The indirect immunofluorescence assay using cardiac tissue from chickens, quails and ducks for identification of influenza A virus during an outbreak of highly pathogenic avian influenza virus (H5N1): a rapid and simple screening tool for limited resource settings

Here we describe the diagnostic utility of the indirect immunofluorescence assay (IFA) during a recent outbreak of highly pathogenic avian influenza (HPAI) subtype H5N1 virus in southern Thailand and demonstrate the usefulness of the cardiac tissue from infected chickens, quail, and ducks for diagnosis. The most reliable sample for IFA diagnosis of influenza A virus was cardiac tissue (83.0%; 44/53) which when divided by species (chicken, quail and duck cardiac tissues) gave respective positivity rates of 88% (22/25), 88.9% (16/18) and 60.0% (6/10). Cardiac tissue also gave the highest IFA intensity for the three species. We believe that the IFA method has wide applicability in developing countries or remote settings where clinically similar avian diseases with high morbidity and mortality such as Newcastle disease and fowl cholera are common and could be rapidly excluded thereby conserving valuable reference laboratory capacity for true HPAI outbreaks.     

Cx43 is involved in electrical remodeling of atrial myocytes through regulating L-type calcium current

AIM: To investigate whether the association of connexin 43(Cx43) and L-type calcium channel involved in the pathogenesis of atrial fibrillation (AF). METHODS: The biochemical assays and whole-cell patch-clamp technique were used to study the expression of Cx43 in human atrial tissue. The co-localization of Cx43 and L -type calcium channel, and the regulation of L-type calcium current in atrial myocytes were investigated. RESULTS: The expression of Cx43 at mRNA and protein levels was decreased in human atrial tissues of AF patients. In cultured atrium-derived myocytes (HL-1 cells), knockdown of Cx43 significantly inhibited the mRNA expression of L-type calcium channel α1c subunit, as well as L-type calcium current. Co-localization of Cx43 with L-type calcium channel α1c subunit in mouse atrial myocytes was observed. CONCLUSION: The decrease in Cx43 is involved in the pathogenesis of AF, probably through reducing the L-type calcium current in atrial myoctyes by co-localization with L-type calcium channel, thus representing the potential pathogenesis in atrial fibrillation.     

肺癌术后并发心律失常的相关因素分析

目的：探讨诱发肺癌术后心律失常的相关因素,为其防治提供依据。方法：对2002年3月～2006年8月期间的肺癌手术患者212例进行回顾性分析,以可能诱发术后心律失常的相关因素分组,比较术后心律失常的发生率。结果：年龄≥60岁、术前肺功能差（FEV1/FVC〈70%）、全肺切除术、术中失血量≥400ml、术后未安装镇痛泵等病例术后心律失常的发生率较高（P〈0.05）。结论：年龄≥60岁、术前肺功能FEV1/FVC〈70%、全肺切除术、术中失血量≥400ml等可能是肺癌术后心律失常的危险因素;术前改善心肺功能,严格掌握手术适应症,术中减少出血、避免损伤自主神经,术后纠正低氧、镇痛等可能会有效减少术后心律失常发生。     

Effects of serum from patients with atrial fibrillation on chemotaxis of cardiac fibroblasts

AIM: To investigate the effects of the serum from the patients with atrial fibrillation (AF) on the chemotaxis of rat cardiac fibroblasts.METHODS: Cardiac fibroblasts were isolated from the ventricles of neonatal Sprague-Dawley rats and primarily cultured with digestion and differential adhesion. The cells in 3 to 4 passages were used for Transwell chamber assay to determine the chemotatic effects of the sera.RESULTS: Compared with control group, the cells that migrated through the polycarbonate membrane were obviously increased in AF group. The strongest chemotaxis was induced by the serum from the patients with persistent atrial fibrillation(pers-AF).The number of migrated cells in non-AF atrial arrhythmia(AA) group was higher than that in paroxysmal atrial fibrillation(paro-AF) group, and that in control group was the lowest. The results of multiple Logistic regression analysis showed that the migrated cells were related to AF and left atrial diameter.CONCLUSION: The chemotactic effect of AF serum is obviously higher than that of control serum. The differences of AF sera among groups show that myocardial fibrosis is a chronic, insidious and delayed process. The migration and infiltration of cardiac fibroblasts indirectly reflect the presence, severity and extent of the myocardial damage. The changes of migrated cells precede the changes of left atrial diameter, indicating that the cause of fibrosis is more important, and the positive correlation between AF and left atrial diameter may not be the direct causality.     

Use of transesophageal atrial pacing to provide temporary chronotropic support in a dog undergoing permanent pacemaker implantation

A 14.5-kg, 13-year-old female spayed Cocker spaniel was evaluated because of episodic hind limb weakness. Results of examination were consistent with sick sinus syndrome with intermittent second-degree atrioventricular block. Transesophageal atrial pacing was successful in providing chronotropic support during permanent pacemaker implantation. Transesophageal atrial pacing appears to be a viable option for temporary atrial pacing in dogs with hemodynamically marked bradycardia without significant atrioventricular blockade.     

A COMPLICATION OF CARDIAC CATHETERIZATION IN THE DOG:

This case history describes a fatal complication of cardiac catheterization in a dog. A 2-year-old intact female miniature Schnauzer presented with a one month history of coughing, tachypnea, and dyspnea that was unresponsive to medical therapy. On clinical examination, a 4/6 systolic murmur was auscultated over the left and right fourth intercostal spaces. Lung sounds were diffusely increased. Survey radiographs revealed cardiomegaly and pulmonary edema. Cardiac catheterization was undertaken to clarify the cause of congestive cardiac failure but was abandoned when contrast medium was seen in the pericardial sac following an attempted injection of the contrast medium into the left ventricle. During recovery from anesthesia progressive pallor, hemothorax, and respiratory distress developed. The dog died 10 hours later despite aggressive support therapy. Gross necropsy revealed hemorrhage into the pericardial sac and pleural space, thrombus formation around and perforation of the right coronary artery.     

Cardiac troponin-I concentration in dogs with cardiac disease

Cardiac troponin-I (cTnI) is a highly sensitive and specific marker of myocardial injury and can be detected in plasma by immunoassay techniques. The purpose of this study was to establish a reference range for plasma cTnI in a population of healthy dogs using a human immunoassay system and to determine whether plasma cTnI concentrations were high in dogs with acquired or congenital heart disease, specifically cardiomyopathy (CM), degenerative mitral valve disease (MVD), and subvalvular aortic stenosis (SAS). In total, 269 dogs were examined by physical examination, electrocardiography, echocardiography, and plasma cTnI assay. In 176 healthy dogs, median cTnI was 0.03 ng/mL (upper 95th percentile = 0.11 ng/mL). Compared with the healthy population, median plasma cTnI was increased in dogs with CM (0.14 ng/mL; range, 0.03-1.88 ng/mL; P < .001; n = 26), in dogs with MVD (0.11 ng/mL; range, 0.01-9.53 ng/mL; P < .001; n = 37), and in dogs with SAS (0.08 ng/mL; range, 0.01-0.94 ng/mL; P < .001; n = 30). In dogs with CM and MVD, plasma cTnI was correlated with left ventricular and left atrial size. In dogs with SAS, cTnI demonstrated a modest correlation with ventricular wall thickness. In dogs with CM, the median survival time of those with cTnI >0.20 ng/mL was significantly shorter than median survival time of those with cTnI <0.20 ng/mL (112 days versus 357 days; P = .006). Plasma cTnI is high in dogs with cardiac disease, correlates with heart size and survival, and can be used as a blood-based biomarker of cardiac disease.     

瓜萎建白半夏汤对大鼠缺血再灌注 心肌细胞凋亡及Bcl-2 , Bax蛋白表达影响

目的研究瓜萎燕白半夏汤对缺血再灌注损伤大鼠心肌细胞凋亡及Bcl-2 , Bax蛋白表达影响〔方法通过 结扎大鼠冠状动脉左前降支造成心肌缺血再灌注损伤模型,各组动物至实验时限心肌缺血30 min、再灌注90 min后, 取出心脏、采用TUNEL检测心肌细胞凋亡,免疫组化方法检测心肌Bcl-2 , Bax蛋白表达、结果与假手术组对照,模型 组细胞凋亡率及Bax表达水平均明显升高、Bcl-2表达水平降低,组间比较差异有统计学意义(P<0.01);瓜萎燕白半夏 能有效降低Bax表达、升高Bcl-2表达水平,抑制细胞凋亡的发生,与模型组比较,差异有统计学意义(P<0.01)、结论瓜 萎燕白半夏汤可能通过上调Bcl-2 ,卜调Bax蛋白表达而有效抑制心肌缺血再灌注损伤大鼠心肌细胞凋亡的发生