目的 探究血清簇集蛋白(Clusterin)、组织激肽释放酶1(KLK1)与老年急性心力衰竭(AHF)病情关系及对患者预后的评估价值.方法 选取2019 年7 月—2021 年10 月河北医科大学第二医院心血管内四科收治老年AHF患者80 例为研究对象(AHF组),另选取同期在医院行健康体检者 80 例为健康对照组.根据心功能将AHF患者分为轻度亚组(n =37)和重度亚组(n =43),依据患者是否发生终点事件分为终点事件亚组(n =36)和非终点事件亚组(n =44).比较各组间的基本资料及血清Clusterin、KLK1 水平;Spearman相关性分析AHF患者血清Clusterin、KLK1 水平与病情程度之间的关系;Logistic 回归分析 AHF 患者发生终点事件的影响因素;受试者工作特征曲线(ROC)分析血清Clusterin、KLK1 对AHF患者预后的评估价值.结果 AHF组SCr、TnI、CK-MB、NT-proBNP、LVEDD、BNP、Clusterin水平显著高于健康对照组(t/P =8.439/<0.001、31.607/<0.001、5.212/<0.001、15.676/<0.001、11.106/<0.001、17.847/<0.001、12.792/<0.001),LVEF、KLK1 水平显著低于健康对照组(t/P =3.900/<0.001、8.963/<0.001);终点事件亚组 TnI、CK-MB、NT-proBNP、BNP、Clusterin 水平均显著高于非终点事件亚组(t/P = 8.389/<0.001、3.517/0.001、10.993/<0.001、5.626/<0.001、7.303/<0.001),LVEF、KLK1 水平显著低于非终点事件亚组(t/P =2.749/0.007、6.225/<0.001);与轻度亚组患者比较,重度亚组患者血清Clusterin水平显著升高,KLK1水平显著降低(t/P =8.236/<0.001、12.703/<0.001);Spearman相关性分析结果显示,AHF患者血清Clusterin水平与病情程度呈显著正相关(r =0.613,P<0.01),而血清KLK1 水平与病情程度呈显著负相关(r =-0.680,P<0.01);Logistic回归分析结果显示,BNP高、Clusterin高、KLK1 低是影响AHF患者发展为不良终点事件的危险因素[OR(95%CI)=1.401(1.132～1.735)、1.545(1.219～1.959)、1.624(1.246～2.116)];ROC曲线结果显示,血清 Clusterin、KLK1 及二者联合评估AHF患者预后的曲线下面积(AUC)分别为0.879、0.834、0.964,二者联合评估AHF患者预后的AUC大于单项预测(Z/P =2.001/0.045、2.712/0.007).结论 血清Clusterin、KLK1 对老年AHF病情及预后具有良好的评估价值.

目的 探讨lncRNA PVT1对缺氧复氧(H/R)诱导的心肌细胞凋亡、氧化应激的影响及其对miR-761的调控作用.方法 采用H/R诱导大鼠心肌细胞H9c2建立细胞损伤模型,分别将si-NC、si-PVT1、miR-mimics-NC、miR-761 mimics、si-PVT1与miR-inhibitor-NC、si-PVT1与miR-761 inhibitor转染至H/R诱导的心肌细胞;采用qRT-PCR法检测PVT1、miR-761的表达量;采用流式细胞术检测细胞凋亡率;采用试剂盒检测丙二醛(MDA)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)的含量;双荧光素酶报告实验检测PVT1与miR-761的靶向关系;Western blot法检测B淋巴细胞瘤2(Bcl-2)、B淋巴细胞瘤2相关蛋白(Bax)的蛋白表达量.结果 与对照组比较,H-/R组PVT1的表达水平升高(P＜0.05),miR-761的表达水平降低(P＜0.05).与H/R+si-NC组比较,H/R+si-PVT1组凋亡率及Bax蛋白水平降低(P＜0.05),Bcl-2蛋白水平及SOD、CAT的活性升高(P＜0.05),MDA的含量降低(P＜0.05).与H/R+miR-mimics-NC组比较,H/R+miR-761 mimics组凋亡率及Bax蛋白水平降低(P＜0.05),Bcl-2蛋白水平及SOD、CAT的活性升高(P＜0.05),MDA的含量降低(P＜0.05).双荧光素酶报告实验证实PVT1可靶向结合miR-761;干扰miR-761可明显逆转沉默PVT1对H/R诱导的心肌细胞氧化应激及凋亡的作用.结论 沉默PVT1可通过上调miR-761而抑制细胞凋亡及氧化应激从而减轻H/R诱导的心肌细胞损伤.

目的 探讨冠心病患者血浆脂蛋白相关磷脂酶A2、血尿酸、高敏C-反应蛋白水平变化.方法 选取2016年09月～2018年11月收治的研究对象为60例冠心病患者,按照冠心病类型分为三组,即为观察1组(急性心肌梗死)、观察2组(不稳定型心绞痛)、观察3组(稳定性型心绞痛),同期选择20例健康体检人员作为研究对象——对照组;且对各组受检人员的脂蛋白相关磷脂酶A2、血尿酸、高敏C-反应蛋白水平进行观察及评估.结果 观察1组患者的血浆脂蛋白相关磷脂酶A2、血尿酸、高敏C-反应蛋白高于观察2、3组及对照组数据,差异有统计学意义(P<0.05).结论 血浆脂蛋白相关磷脂酶A2、血尿酸及高敏C-反应蛋白水平变化可用于预测冠心病,以便于为治疗提供正确方向,与此同时还能反映冠状动脉进程.

Objectives: To explore that if mid-regional sequence of pro-A-type natriuretic peptide (MR-proANP) may have a good value of diagnosis in heart failure with preserved ejection fraction (HFpEF) compared with N-terminal pro-B-type natriuretic peptide (NT-proBNP). Methods: Participants with cardiovascular disease who were enrolled in this study were divided into the nonheart failure (non-HF) group (n=75), HFpEF group (n=65), and HF with reduced ejection fraction (HFrEF) group (n=50). The MR-proANP and NT-proBNP levels in plasma from all patients were measured by enzyme-linked immunosorbent assay. Results: The plasma levels of MR-proANP and NT-proBNP in HFpEF and HFrEF groups were higher than those in non-HF group (P<.05). MR-proANP levels were significantly different (P<.05) in different New York Heart Association class patients with HFpEF. In the diagnostic analysis area under the curve of MR-proANP (0.844) was higher than that of NT-proBNP (0.518, P<.001). The left atrial volume index in the HFrEF group was higher than HFpEF group (P<.05); however, both of these groups had a higher index than non-HF group (P<.05). Conclusion: Results indicated that MR-proANP may be more sensitive and specific than NT-proBNP in diagnosing HFpEF. It may be used as a potential diagnostic biomarker in patients with HFpEF.

The purpose of this study was to investigate whether atrial overexpression of angiotensin-converting enzyme 2 (ACE2) by homogeneous transmural atrial gene transfer can reverse atrial remodeling and its mechanisms in a canine atrial-pacing model. Twenty-eight mongrel dogs were randomly divided into four groups: Sham-operated, AF-control, gene therapy with adenovirus-enhanced green fluorescent protein (Ad-EGFP) and gene therapy with Ad-ACE2 (Ad-ACE2) (n = 7 per subgroup). AF was induced in all dogs except the Sham-operated group by rapid atrial pacing at 450 beats/min for 2 weeks. Ad-EGFP and Ad-ACE2 group then received epicardial gene painting. Three weeks after gene transfer, all animals except the Sham group underwent rapid atrial pacing for another 3 weeks and then invasive electrophysiological, histological and molecular studies. The Ad-ACE2 group showed an increased ACE2 and Angiotensin-(1–7) expression, and decreased Angiotensin II expression in comparison with Ad-EGFP and AF-control group. ACE2 overexpression attenuated rapid atrial pacing-induced increase in activated extracellular signal-regulated kinases and mitogen-activated protein kinases (MAPKs) levels, and decrease in MAPK phosphatase 1(MKP-1) level, resulting in attenuation of atrial fibrosis collagen protein markers and transforming growth factor-β1. Additionally, ACE2 overexpression also modulated the tachypacing-induced up-regulation of connexin 40, down-regulation of connexin 43 and Kv4.2, and significantly decreased the inducibility and duration of AF. ACE2 overexpression could shift the renin–angiotensin system balance towards the protective axis, attenuate cardiac fibrosis remodeling associated with up-regulation of MKP-1 and reduction of MAPKs activities, modulate tachypacing-induced ion channels and connexin remodeling, and subsequently reduce the inducibility and duration of AF.

目的探讨血管紧张素（-1-7）[Angiotensin（-1-7）,Ang（-1-7）]对血管紧张素Ⅱ（AngiotensinⅡ,AngⅡ）在大鼠心脏成纤维细胞（Cardiac fibroblasts,CFs）中信号转导的影响及其机制。方法原代分离培养并鉴定新生SD大鼠的CFs,将细胞分为8组:空白对照组、AngⅡ组、Ang（-1-7）组、AngⅡ+Ang（-1-7）组、AngⅡ+Ang（-1-7）+A-779组、AngⅡ+Ang（-1-7）+PAO组、Ang（-1-7）+PAO组和AngⅡ+PAO组,Western blot法检测细胞外信号调节激酶1/2（Extracellular signal regulated kinase 1/2,ERK1/2）和磷酸化的ERK1/2（p-ERK1/2）的表达;免疫沉淀捕捉分析法检测蛋白酪氨酸磷酸酶-1（Src-homology domain 2 containing protein tyrosine phosphatase-1,SHP-1）的酶活性;实时荧光定量PCR检测转化生长因子-β1（Transforming growth factor-β1,TGF-β1）、I型胶原蛋白（CollagenⅠ,ColⅠ）和Ⅲ型胶原蛋白（CollagenⅢ,ColⅢ）基因mRNA的转录水平。结果 AngⅡ可增加细胞内p-ERK1/2的表达水平和p-ERK/ERK值（磷酸化的p-ERK1/2与总的ERK1/2的比值）,Ang（-1-7）通过与Mas受体结合可拮抗AngⅡ引起的上述效应;Ang（-1-7）通过与Mas受体结合而激活胞内SHP-1的活性,并可拮抗AngⅡ所致的SHP-1活性降低;抑制SHP-1的活性后,Ang（-1-7）拮抗AngⅡ诱导的p-ERK1/2表达水平和p-ERK/ERK值增高的效应被抑制。Ang（-1-7）对TGF-β1、ColⅠ和ColⅢ基因mRNA的转录水平无显著影响,但可抑制AngⅡ所诱导的上述基因mRNA转录水平的增加。结论 Ang（-1-7）通过与Mas受体结合而激活SHP-1,该效应与Ang（-1-7）拮抗AngⅡ诱导的p-ERK1/2的表达水平和p-ERK/ERK值增高密切相关;Ang（-1-7）可抑制AngⅡ所诱导的TGF-β1、ColⅠ和ColⅢ基因表达上调,这些现象可能体现了一种Ang（-1-7）拮抗AngⅡ所致的不良效应的保护性机制。

在肾素-血管紧张素系统(RAS)中,血管紧张素转化酶2(ACE2)与血管紧张素(AngⅡ)之间的代谢关系在ACE2-Ang(1-7)-Mas轴保护心房颤动(房颤)的心肌细胞作用中发挥着重要作用。作为ACE的唯一同系物,ACE2可以水解AngⅡ为七肽Ang(1-7),而Ang(1-7)可以特异性激活Mas受体,发挥抗纤维化、抗增殖、抗炎、扩血管及保护内皮等作用,从多方面拮抗RAS系统中AngⅡ的病理性作用。目前研究表明,ACE2-Ang(1-7)-Mas轴中ACE2及Ang(1-7)可以分别通过抑制促分裂原活化蛋白激酶信号通路从而阻断AngⅡ的病理过程,发挥保护心肌细胞的功能。基于目前对房颤治疗的局限性和房颤发病机制的逐渐认识,ACE2-Ang(1-7)-Mas轴对心血管病理过程特别是房颤发生的逆转作用逐渐得到重视,该轴的心肌保护作用有可能成为房颤治疗的一个重要分支。

Gene transfer in atria myocardium with high efficiency has been technically challenging.Developing new methods for atrial gene transfer can be difficult in many aspects,such as the shortage of blood supply and the thinness of atrial wall.However,myocardial gene therapy remained to be an important approach for the investigation and treatment of cardiac disease.We explored the feasibility of casting mixture solution on the atrial epicardial to improve the efficiency and targeting in canine myocardial gene transfer.The mixture solution contained rAd-EGFP(1×109 pfu/mL),trypsin(0.5%)and poloxamer F407(35%) and was spread on the right atrial epicardial via median sternotomy.The fluorescence intensity of atrial tissue at different transfer time points was observed by fluorescent microscopy.Real time PCR was used to analysis EGFP mRNA expression level positions from different part of the heart.HE straining was used to estimate the inflammatory and anatomical changes.Masson's trichrome straining was used to examine fibrosis.The EGFP mRNA expression showed an initial increase followed by a decrease with a peak at week 3,where almost every myocardial layers of right atrium anterior wall(RAAW) showed the transgene expression with the highest level at the right atrium anterior(RAA).Similar level of EGFP expression was observed in RAAW and RAPW;and the expression in interatrial septum(IAS) was significantly lower than right atrium(RA),but higher than that in left atrium(LA).The lowest levels of EGFP mRNA expression was found in ventricular and other organs such as lung,liver,spleen,skeletal muscle,stomach,and small intestine,etc.(* P<0.01 compared with RA).No obvious inflammatory and fibrosis changes were found in RA tissues.Our gene transfer method by casting mixture solution on atrial epicardial showed potentials of high efficiency,targeting ability and safety.

肾素-血管紧张素系统(RAS)在机体维持心血管正常功能及电解质平衡中发挥着关键作用.过度激活RAS后,将会引起身体的一系列病理改变,而RAS的重要组成部分--血管紧张素(Ang)Ⅱ通过激活血管紧张素Ⅱ(AT)1型受体在介导心房颤动(房颤)的发生和发展中的病理作用已被广泛认同.随着血管紧张素转换酶(ACE)的同系物ACE2的被发现,AngⅡ的代谢调节方式又出现了新的路径.

在肾素-血管紧张素系统（RAS）中,血管紧张素转化酶2（ACE2）与血管紧张素（AngⅡ）之间的代谢关系在ACE2-Ang（1-7）-Mas轴保护心房颤动（房颤）的心肌细胞作用中发挥着重要作用。作为ACE的唯一同系物,ACE2可以水解AngⅡ为七肽Ang（1-7）,而Ang（1-7）可以特异性激活Mas受体,发挥抗纤维化、抗增殖、抗炎、扩血管及保护内皮等作用,从多方面拮抗RAS系统中AngⅡ的病理性作用。目前研究表明,ACE2-Ang（1-7）-Mas轴中ACE2及Ang（1-7）可以分别通过抑制促分裂原活化蛋白激酶信号通路从而阻断AngⅡ的病理过程,发挥保护心肌细胞的功能。基于目前对房颤治疗的局限性和房颤发病机制的逐渐认识,ACE2-Ang（1-7）-Mas轴对心血管病理过程特别是房颤发生的逆转作用逐渐得到重视,该轴的心肌保护作用有可能成为房颤治疗的一个重要分支。