肌浆网钙ATP酶基因转导对慢性心力衰竭犬心肌蛋白质组影响的初步研究

目的 分析心肌肌浆网Ca2+-ATP酶(sarcoplasmic reticulum Ca2+ ATPase 2a,SERCA2a)基因转导对慢性心力衰竭(HF)犬心肌蛋白质组的影响,探讨SERCA2a基因转导改善心功能的机制.方法 快速右心室起搏建立HF犬模型并随机分为HF组、HF+绿色荧光蛋白(enhanced green fluorescent pmtein,EGFP)组、HF+SERCA2a组.后两组分别向心肌内注射携带EGFP和SERCA2a基因的rAAV载体.于基因转导30 d时停止起搏后进行超声心动图和血流动力学检查并制备心室肌双向电泳蛋白样品和心肌双向电泳图谱,图像分析软件分析蛋白表达差异点,MALDI-TOF-MS数据库搜索鉴定蛋白质.结果 基因转导30 d时,HF+SERCA2a组犬的症状、超声心动图和血流动力学指标与HF+EGFP组相比有显著好转(P＜0.05);与对照组相比差异无统计学意义(P＞0.05).挑选SERCA2a基因转导后表达量发生明显改变的10个蛋白点进行分析,经质谱鉴定分别为心肌收缩相关蛋白、线粒体能量代谢酶类和应激相关蛋白.结论 以rAAV为载体介导SERCA2a基因转导能够改善HF犬心脏的收缩和舒张功能,其可能的机制是恢复了心肌收缩相关蛋白正常表型或正常表达量,增加了心肌能量的产生,改变了应激相关蛋白的表达.     

Clinical Importance of Left Atrial Infiltration in Cardiac Transthyretin Amyloidosis

ObjectivesThe aim of this study was to characterize left atrial (LA) pathology in explanted hearts with transthyretin amyloid cardiomyopathy (ATTR-CM); LA mechanics using echocardiographic speckle-tracking in a large cohort of patients with ATTR-CM; and to study the association with mortality.BackgroundThe clinical significance of LA involvement in ATTR-CM is of great clinical interest.MethodsCongo red staining and immunohistochemistry was performed to assess the presence, type, and extent of amyloid and associated changes in 5 explanted ATTR-CM atria. Echo speckle tracking was used to assess LA reservoir, conduit, contractile function, and stiffness in 906 patients with ATTR-CM (551 wild-type (wt)-ATTR-CM; 93 T60A-ATTR-CM; 241 V122I-ATTR-CM; 21 other).ResultsThere was extensive ATTR amyloid infiltration in the 5 atria, with loss of normal architecture, vessels remodeling, capillary disruption, and subendocardial fibrosis. Echo speckle tracking in 906 patients with ATTR-CM demonstrated increased atrial stiffness (median [25th-75th quartile] 1.83 [1.15-2.92]) that remained independently associated with prognosis after adjusting for known predictors (lnLA stiff: HR: 1.23; 95% CI: 1.03-1.49; P = 0.029). There was substantial impairment of the 3 phasic functional atrial components (reservoir 8.86% [5.94%-12.97%]; conduit 6.5% [4.53%-9.28%]; contraction function 4.0% [2.29%-6.56%]). Atrial contraction was absent in 22.1% of patients whose electrocardiograms showed sinus rhythm (SR) “atrial electromechanical dissociation” (AEMD). AEMD was associated with poorer prognosis compared with patients with SR and effective mechanical contraction (P = 0.0018). AEMD conferred a similar prognosis to patients in atrial fibrillation.ConclusionsThe phenotype of ATTR-CM includes significant infiltration of the atrial walls, with progressive loss of atrial function and increased stiffness, which is a strong independent predictor of mortality. AEMD emerged as a distinctive phenotype identifying patients in SR with poor prognosis.     

Effect of clot removal on cerebrovascular contraction after subarachnoid hemorrhage in the monkey: Pharmacological study

Summary Clot removal at early surgery has been reported to be clinically effective for the prevention of cerebral vasospasm following subarachnoid hemorrhage (SAH) due to rupture of an intracranial aneurysm. We examined the most efficacious timing of mechanical clot removal on pharmacological responses in a monkey SAH model. Cynomolgus monkeys (Macaca fascicularis) were randomized into five groups: sham-operated, clot removal in which the clot was removed 48, 72, or 96h after SAH, and clot groups. An autologous blood clot was placed around the bilateral major cerebral arteries after craniectomy to mimic the hemorrhage. Seven days after the SAH, proximal and successively distal parts of the middle cerebral arteries were cut into rings for isometric tension measurement. The contractile responses to potassium chloride, 5-hydroxytryptamine, norepinephrine, adenosine triphosphate, prostaglandin F₂, and hemoglobin were greater in the proximal parts than in the distal parts in each group. Compared with the sham-operated group, the responses of the clot-removal and clot groups to the drugs were progressively attenuated. The maximum responses to 5-hydroxytryptamine in the proximal parts and to adenosine triphosphate in the distal parts started to decrease, significantly, in the clot-removal group 48h after SAH, while most of the responses to the other agonists began to decrease in the clot-removal groups later than 72h after SAH. These results suggest that the attenuation of cerebrovascular contractile responses 7 days after SAH is pharmacologically inevitable, even if the clot is removed as early as 48h after the SAH. Clot removal may thus be recommended within 48h after SAH to ameliorate the severity of cerebral vasospasm following SAH.     

Electrocardiographic characteristics of repetitive monomorphic right ventricular tachycardia originating near the His-bundle

INTRODUCTION: Most idiopathic nonreentrant ventricular tachycardia (VT) and ventricular premature contractions (VPCs) arise from the right or left ventricular outflow tract (OT). However, some right ventricular (RV) VT/VPCs originate near the His-bundle region. The aim of this study was to investigate ECG characteristics of VT/VPCs originating near the His-bundle in comparison with right ventricular outflow tract (RVOT)-VT/VPCs. METHODS AND RESULTS: Ninety RV-VT/VPC patients underwent catheter mapping and radiofrequency ablation. ECG variables were compared between VT/VPCs originating from the RVOT and near the His-bundle. Ten patients had foci near the His-bundle (HIS group), with the His-bundle local ventricular electrogram preceding the QRS onset by 15-35 msec (mean: 22 msec) and His-bundle pacing produced a nearly identical ECG to clinical VT/VPCs. The HIS group R wave amplitude in the inferior leads (lead III: 1.0 +/- 0.6 mV) was significantly lower than that of the RVOT group (1.7 +/- 0.4 mV, P < 0.05). An R wave in aVL was present in 6 of 10 HIS group patients, while almost all RVOT group patients had a QS pattern in aVL. Lead I in HIS group exhibited significantly taller R wave amplitudes than RVOT group. HIS group QRS duration in the inferior leads was shorter than that of the RVOT group. Eight of 10 HIS group patients exhibited a QS pattern in lead V1 compared to 14 of 81 RVOT group patients. HIS group had larger R wave amplitudes in leads V5 and V6 than RVOT group. CONCLUSION: VT/VPCs originating near the His-bundle have distinctive ECG characteristics. Knowledge of the characteristic QRS morphology may facilitate catheter mapping and successful ablation.     

Dissecting the Roles of Supervised and Unsupervised Learning in Perceptual Discrimination Judgments

Our ability to compare sensory stimuli is a fundamental cognitive function, which is known to be affected by two biases: choice bias, which reflects a preference for a given response, and contraction bias, which reflects a tendency to perceive stimuli as similar to previous ones. To test whether both reflect supervised processes, we designed feedback protocols aimed to modify them and tested them in human participants. Choice bias was readily modifiable. However, contraction bias was not. To compare these results to those predicted from an optimal supervised process, we studied a noise-matched optimal linear discriminator (Perceptron). In this model, both biases were substantially modified, indicating that the “resilience” of contraction bias to feedback does not maximize performance. These results suggest that perceptual discrimination is a hierarchical, two-stage process. In the first, stimulus statistics are learned and integrated with representations in an unsupervised process that is impenetrable to external feedback. In the second, a binary judgment, learned in a supervised way, is applied to the combined percept.SIGNIFICANCE STATEMENT The seemingly effortless process of inferring physical reality from the sensory input is highly influenced by previous knowledge, leading to perceptual biases. Two common ones are contraction bias (the tendency to perceive stimuli as similar to previous ones) and choice bias (the tendency to prefer a specific response). Combining human psychophysical experiments with computational modeling we show that they reflect two different learning processes. Contraction bias reflects unsupervised learning of stimuli statistics, whereas choice bias results from supervised or reinforcement learning. This dissociation reveals a hierarchical, two-stage process. The first, where stimuli statistics are learned and integrated with representations, is unsupervised. The second, where a binary judgment is applied to the combined percept, is learned in a supervised way.     

Validity of a portable hanging scale to assess the isometric muscle strength during knee movement

Inexpensive and practical equipment to assess muscle strength can disseminate objective measures, which can provide valid information to implement effective treatment and exercise training. The purpose of the study was to examine the instrumental validity of the hanging scale (HS) to assess the muscle strength during knee flexion and extension by comparing the peak force values to a standard equipment (laboratory-grade load cell [LLC]). Knee isometric strength was assessed on thirty-two subjects (16 women and 16 men, 22 [5] years, 171 [9] cm, 69 [15] kg, 23 [4] kg/m²). The knee flexion was tested by placing the volunteer's body in prone with the knee flexed at 90°. Knee extension was assessed with the volunteer seated on a chair with the feet resting on the floor, knees, and hips flexed at 90°. No differences were observed comparing the isometric peak forces between a laboratory-grade load cell and a HS (p > 0.05). The intraclass correlation coefficient (ICC)_1,1 returned values above 0.90. The Cronbach's α test also returned values above 0.90 for all comparisons. Bland-Altman results showed high levels of agreement with low risk of bias. The HS appears to be a valid method to assess the knee isometric strength. The portability, the cost-effectiveness and the simple user-friendly system provides an effective way to assess the knee isometric strength.     

Collagen gel contraction as a measure of fibroblast function in an animal model of subsynovial connective tissue fibrosis

Carpal tunnel syndrome (CTS) is a peripheral neuropathy characterized by non‐inflammatory fibrosis of the subsynovial connective tissues (SSCT). A rabbit model of CTS was developed to test the hypothesis that SSCT fibrosis causes the neuropathy. We used a cell‐seeded collagen‐gel contraction model to characterize the fibrosis in this model in terms of cellular mechanics, specifically to compare the ability of SSCT cells from the rabbit model and normal rabbits to contract the gel, and to assess the effect of transforming growth factor‐β1,which is upregulated in CTS, on these cells. SSCT fibrosis was induced in six retired breeder female rabbits which were sacrificed at 6 weeks (N = 3) and 12 weeks (n = 3). An additional two rabbits served as controls. SSCT was harvested according to a standard protocol. Gels seeded with SSCT cells from rabbits sacrificed at 6 weeks had significantly higher tensile strength (p < 0.001) and Young's modulus (p < 0.001) than gels seeded with cells from rabbits sacrificed at 12 weeks or control animals. TGF‐β1 significantly increased the decay time constant (p < 0.001), tensile strength (p < 0.001), and Young's modulus (p < 0.001) regardless of the cell source. This model may be useful in screening therapeutic agents that may block SSCT fibrosis, identifying possible candidates for CTS treatment. © 2015 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 33:668–674, 2015.     

起源点邻近二尖瓣环的频发室性早搏体表心电图特征及射频消融治疗

目的评估起源点邻近二尖瓣环的频发室性早搏(简称室早)体表心电图特点及射频消融治疗效果。方法10例频发室早患者接受常规电生理检查及射频消融治疗,对所有病例12导联体表心电图进行分析。结果10例室早均消融成功,并证实起源点邻近二尖瓣环的不同部位,根据成功消融靶点将本组病例分为3组,二尖瓣环前侧壁组(4例)、二尖瓣环后侧壁组(3例)、二尖瓣环后间隔组(3例)。所有病例胸前导联R波移行区位于V1～V2导联,绝大部分病例V6导联可见s波。对各组二尖瓣环室早心电图做进一步比较,可概括出系列心电图判断指标用以估计消融靶点的部位。结论射频消融治疗起源点邻近二尖瓣环的频发室早可取得良好效果,掌握其体表心电图特点有助于判定室早的起源部位。     

Cyclic adenosine monophosphate modulation of contractility via slow Ca2+ channels in chick heart

The catecholamines exert a positive inotropic effect associated with elevated tissue cyclic AMP levels and possibly with increase in the number of membrane slow cationic channels available for voltage activation. In the present study, catecholamines (isoproterenol, dopamine and dobutamine) were tested for their ability to affect the maximum upstroke velocity (+ V_max) of the slow action potentials, the first derivative ($\text{d}\text{T}\text{d}\text{t}$) of developed tension accompanying the slow responses, and the tissue cyclic AMP levels in the ventricular myocardium of isolated perfused chick hearts. To study the slow channels exclusively, the fast Na⁺ channels were voltage inactivated by elevated (25 mm) K⁺. In this condition of functional removal of the fast channels, the heart could not be excited by intense electrical stimulation. It was found that these catecholamines induced slow action potentials accompanied by contractions. Elevation of the concentration of these agents produced increases in + V_max, $\text{d}\text{T}\text{d}\text{t}$, and cyclic AMP in a dose-dependent fashion; a close correlation was obtained between the cyclic AMP level, + V_max and $\text{d}\text{T}\text{d}\text{t}$. These results support the hypothesis that the increases in + V_max of the slow action potentials and in contraction are explained by increase in the number of available slow channels mediated by intracellular cyclic AMP levels, and the resulting increase in the Ca²⁺ influx.     

Role of cholecystokinin in the regulation of liquid gastric emptying and gastric motility in humans: studies with the CCK antagonist loxiglumide

Background—Exogenous cholecystokinin (CCK)inhibits antral motility and slows gastric emptying (GE) but the effectof endogenous CCK on the gastric motor mechanisms responsible for GEremains unclear.
Methods—The effect of the CCK-A antagonistloxiglumide (LOX) on GE and motility was studied using magneticresonance imaging in six healthy volunteers after ingestion of 500 mlIntralipid 10% (550 kcal). Subjects were studied in the supineposition on two occasions during intravenous infusion of LOX(66 µmol/kg/h for 10 min followed by 22 µmol/kg/h) or placebo. GEwas determined every 15 minutes using transaxial abdominal scans andmotility was studied by means of 120 coronal scans, 1.2 seconds apart. For each coronal image the proximal and distal (antral) diameters weremeasured at a fixed point in the stomach to determine contraction frequency (ACF) and amplitude (AMP).
Results—GE was faster during LOX infusion thanplacebo (t_1/2 31 (22) versus 115 (67) minutes, p<0.03).There was little variation in the diameter of the proximal stomach witheither LOX or placebo. In the distal stomach marked contractileactivity was observed during LOX (ACF 2.9 (0.2) versus 1.5 (2.9) duringplacebo, p<0.01). AMP also increased during LOX compared with placebo(56 (22)% versus 27 (16)%, p<0.001).
Conclusion—The increases in antral motility arelikely to contribute to the acceleration of GE and suggest that CCK mayregulate GE by acting on the distal stomach although an effect on theproximal stomach cannot be excluded.

Keywords:loxiglumide; magnetic resonance imaging; gastricemptying; gastric motility; antral contraction