冠心病相关循环miRNA

冠心病诊断中最主要的挑战就是从常规的血液样本中鉴定出可靠的临床生物标志物.循环miRNA是一种可以稳定存在于体液中的小分子RNA,具有较高的组织、疾病特异性及敏感性,具有作为新的冠心病非侵入性生物标志物的潜力.通过综述血液样本（全血、血浆、血清、外周血单核细胞（PBMC）、内皮祖细胞（EPC）及血小板）中冠心病相关循环miRNA,及探讨循环miRNA研究中存在的一些问题,为未来筛选出真正具有临床应用价值的循环miRNA生物标志物奠定基础.     

川崎病患儿并发冠状动脉损伤的危险因素及预测模型的构建与评价

摘要 目的：分析川崎病患儿并发冠状动脉损伤（CAL）的危险因素,并构建和评价川崎病患儿并发CAL的预测模型。方法：选取2019年1月～2022年5月我院收治的342例川崎病患儿,根据是否并发CAL分为CAL组和非CAL组。收集所有患儿临床资料,采用单因素和多因素Logistic回归分析川崎病患儿并发CAL的影响因素,并构建预测模型,H-L检验和受试者工作特征（ROC）曲线检验预测模型拟合优度和对川崎病患儿并发CAL的预测价值。结果：342例川崎病患儿CAL发生率为16.67％（57/342）。单因素分析显示,CAL组发热持续时间≥10 d、静脉注射免疫球蛋白（IVIG）治疗延迟、IVIG无反应比例和单核细胞比例（MO％）、嗜酸性粒细胞比例（EO％）、C反应蛋白（CRP）、红细胞沉降率（ESR）、降钙素原（PCT）、心肌肌钙蛋白I（cTnI）、肌酸激酶同工酶（CK-MB）水平高于非CAL组（P均＜0.05）。多因素Logistic回归分析显示,发热持续时间≥10 d、IVIG治疗延迟、IVIG无反应和MO％、CRP、ESR、PCT、cTnI升高为川崎病患儿并发CAL的独立危险因素（P均＜0.05）。H-L检验川崎病患儿并发CAL的预测模型拟合效果良好。ROC曲线分析显示,该模型预测川崎病患儿并发CAL的曲线下面积（AUC）为0.911（95％CI：0.876～0.939）。结论：发热持续时间≥10 d、IVIG治疗延迟、IVIG无反应和MO％、CRP、ESR、PCT、cTnI升高为川崎病患儿并发CAL的危险因素,根据危险因素构建的川崎病患儿并发CAL预测模型价值较高。     

Coronary Artery Disease with Single Coronary Artery

The authors have reviewed the literature in search of the coexistence of single coronary artery with significant coronary artery disease. Two cases of single right coronary artery are described. In both, the anomalies were unsuspected and diagnosed roentgenographically in life. Both patients had angina pectoris, positive graded-exercise stress tests, and hemodynamically significant obstruction or occlusion to the coronary arteries. In neither case was the stenosis proximal or amenable to bypass surgery.     

Gene-Gene Associations with the Susceptibility of Kawasaki Disease and Coronary Artery Lesions

Kawasaki disease (KD) is a systemic vasculitis primarily affecting children < 5 years old. Genes significantly associated with KD mostly involve cardiovascular, immune, and inflammatory responses. Recent studies have observed stronger associations for KD risk with multiple genes compared to individual genes. Therefore, we investigated whether gene combinations influenced KD susceptibility or coronary artery lesion (CAL) formation. We examined 384 single-nucleotide polymorphisms (SNPs) for 159 immune-related candidate genes in DNA samples from KD patients with CAL (n = 73), KD patients without CAL (n = 153), and cohort controls (n = 575). Individual SNPs were first assessed by univariate analysis (UVA) and multivariate analysis (MVA). We used multifactor dimensionality reduction (MDR) to examine individual SNPs in one-, two-, and three-locus best fit models. UVA identified 53 individual SNPs that were significantly associated with KD risk or CAL formation (p < 0.10), while 35 individual SNPs were significantly associated using MVA (p ≤ 0.05). Significant associations in MDR analysis were only observed for the two-locus models after permutation testing (p ≤ 0.05). In logistic regression, combined possession of PDE2A (rs341058) and CYFIP2 (rs767007) significantly increased KD susceptibility (OR = 3.54; p = 4.14 x 10⁻⁷), while combinations of LOC100133214 (rs2517892) and IL2RA (rs3118470) significantly increased the risk of CAL in KD patients (OR = 5.35; p = 7.46 x 10⁻⁵). Our results suggest varying gene-gene associations respectively predispose individuals to KD risk or its complications of CAL.     

血清网膜素-1 与绝经后女性冠心病的相关性

目的：网膜素是最近发现的脂肪因子,肥胖或2 型糖尿病（diabetes mellitus, DM）患者血清网膜素-1 较正常者明显降低。本 次研究主要为观察绝经后女性血清网膜素-1 水平与冠心病的相关性。方法：选取我院心内科住院有心绞痛症状,并行冠脉造影的 105 例绝经后女性患者。依据冠脉造影结果分为冠心病组(67 例)和对照组例（3）,常规收集临床资料,包括年龄、体重指数（body mass index, BMI）、吸烟史、高血压病史、糖尿病史及血液生化和血脂指标;酶联免疫吸附剂测定(enzyme linked immunosorbent assay, ELISA)法检测血清网膜素-1 浓度。结果：冠心病组血清网膜素-1 水平显著低于对照组（205.62± 73.31 vs 401.64± 146.79, P<0.001）。单因素logistic回归分析示吸烟、高血压病史、糖尿病史、高脂血症史、网膜素-1 水平降低是冠心病组的独立危险因素 (P<0.05)。多因素logistic 回归分析示血清网膜素-1 水平降低是冠心病组的独立危险因素(P<0.001)。结论：绝经后女性血清网膜素 -1 水平下降是冠心病的独立危险因素,可能可成为绝经后女性冠心病的预测指标。     

血清网膜素-1与绝经后女性冠心病的相关性

目的：网膜素是最近发现的脂肪因子,肥胖或2型糖尿病（diabetesmellitus,DM）患者血清网膜素-1较正常者明显降低。本次研究主要为观察绝经后女性血清网膜素-1水平与冠心病的相关性。方法：选取我院心内科住院有心绞痛症状,并行冠脉造影的105例绝经后女性患者。依据冠脉造影结果分为冠心病组（67例）和对照组例（3）,常规收集临床资料,包括年龄、体重指数（bodvmassindex,BMI）、吸烟史、高血压病史、糖尿病史及血液生化和血脂指标;酶联免疫吸附剂测定（enzymelinkedimmunosorbentassay,ELISA）法检测血清网膜素-1浓度。结果：冠心痛组血清网膜素-1水平显著低于对照组（205．62±73．31vs401．64±146．79．P〈0．001）。单因素logistic回归分析示吸烟、高血压痛史、糖尿病史、高脂血症史、网膜素-1水平降低是冠心病组的独立危险因素（P〈0．05）。多因素logistic回归分析示血清网膜素-1水平降低是冠心病组的独立危险因素（P〈0．001）。结论：绝经后女性血清网膜素-1水平下降是冠心痛的独立危险因素,可能可成为绝经后女性冠心病的预测指标。     

The IL-33-ST2L Pathway Is Associated with Coronary Artery Disease in a Chinese Han Population

The effects of interleukin-33 (IL-33) on the immune system have been clearly demonstrated; however, in cardiovascular diseases, especially in coronary artery disease (CAD), these effects have not yet been clarified. In this study, we investigate the genetic role of the IL-33-ST2L pathway in CAD. We performed three-stage case-control association analyses on a total of 4,521 individuals with CAD and 4,809 controls via tag SNPs in the genes encoding IL-33 and ST2L-IL-1RL1. One tag SNP in each gene was significantly associated with CAD (rs7025417^T in IL33, p_adj = 1.19 × 10⁻²⁸, OR = 1.39, 95% CI: 1.31–1.47; rs11685424^G in IL1RL1, p_adj = 6.93 × 10⁻³⁰, OR = 1.40, 95% CI: 1.32–1.48). Combining significant variants in two genes, the risk for CAD increased nearly 5-fold (p_adj = 8.90 × 10⁻²¹, OR = 4.98, 95% CI: 3.56–6.97). Traditional risk factors for CAD were adjusted for the association studies by SPSS with logistic regression analysis. With the two variants above, both located within the gene promoter regions, reporter gene analysis indicated that the rs7025417 C>T and rs11685424 A>G changes resulted in altered regulation of IL33 and IL1RL1 gene expression, respectively (p < 0.005). Further studies revealed that the rs7025417 genotype was significantly associated with plasma IL-33 levels in the detectable subjects (n = 227, R² = 0.276, p = 1.77 × 10⁻¹⁷): the level of IL-33 protein increased with the number of rs7025417 risk (T) alleles. Based on genetic evidence in humans, the IL-33-ST2L pathway appears to have a causal role in the development of CAD, highlighting this pathway as a valuable target for the prevention and treatment of CAD.     

A Replication Study for Association of ITPKC and CASP3 Two-Locus Analysis in IVIG Unresponsiveness and Coronary Artery Lesion in Kawasaki Disease

Single-nucleotide polymorphisms (SNPs) in inositol 1,4,5-trisphosphate 3-kinase C (ITPKC, rs28493229) and caspase-3 (CASP3, rs113420705) are associated with susceptibility to KD in Japanese and Taiwanese populations. This study was conducted to investigate the involvement of these 2 SNPs in the risk for intravenous immunoglobulin (IVIG) resistance and coronary artery lesion (CAL) in Taiwanese population. A total of 340 KD patients were subjected to assess by the identification of 2-locus genes model. A combinatorial association between ITPKC (rs28493229) and CASP3 (rs113420705) was found in CAL formation (P = 0.0227, OR: 3.06). KD patients with high-risk genotype had a trend of overrepresentation in IVIG resistance compared with individual SNPs. Our findings suggest the existence of genetic factors affecting patients’ risk for CAL formation and IVIG responsiveness in a Taiwanese population.     

冠心病合并2型糖尿病的冠脉病变特征及危险因素分析

目的:探讨冠心病合并2型糖尿病的冠状动脉病变特征及其相关危险因素.方法:选择2010年1月至2012年1月我院经冠状动脉造影确诊为冠心病合并2型糖尿病的患者227例(DM组)和同期不合并2型糖尿病的冠心病患者229例(NDM组)为研究对象,回顾性分析其血脂、血糖及冠状动脉造影结果,比较两组患者冠状动脉病变的特点,探讨血糖水平对糖尿病合并冠心病患者冠状动脉病变的影响.结果:DM组患者总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)显著高于NDM组(P＜0.05),高密度脂蛋白胆固醇(HDL-C)显著低于NDM组(P＜0.05);DM组患者三支血管病变、弥漫性病变以及狭窄程度大于75％的血管的病例数百分率显著高于NDM组(P＜0.05);在DM患者中,血糖水平控制理想组(A组)的冠状动脉血管狭窄程度大于75％以及发生弥漫性病变的病例数百分率均显著低于血糖控制较差组(B组,P＜0.05).结论:2型糖尿病合并冠心病患者冠状动脉多表现为弥漫和多支病变,狭窄程度严重;血糖和血脂水平异常是其冠脉病变的危险因素;控制患者的血糖水平于正常范围可改善其冠状动脉病变程度并减小其病变范围.     

Oxidative Stress is Associated with Genetic Polymorphisms in One-Carbon Metabolism in Coronary Artery Disease

In view of growing body of evidence favouring the association of aberrations in one-carbon metabolism and oxidative stress in the aetiology of coronary artery disease (CAD), we investigated the risk associated with polymorphisms regulating the folate uptake and transport such as the glutamate carboxypeptidase II (GCPII) C1561T, reduced folate carrier 1 (RFC1) G80A and cytosolic serine hydroxymethyltransferase (cSHMT) C1420T. We further evaluated the impact of seven putatively functional polymorphisms of this pathway on oxidative stress markers. Genotyping was performed on 288 CAD cases and 266 healthy controls along with the dietary folate assessment. GCPII C1561T polymorphism was found to be an independent risk factor (OR 2.71, 95% CI 1.47–4.98) for CAD, whereas cSHMT C1420T conferred protection (OR 0.51, 95% CI 0.37–0.70). Oxidative stress markers like the plasma levels of malondialdehyde, protein carbonyls and 8-oxo-deoxyguanosine were significantly increased and total glutathione was significantly decreased in CAD cases. Elevated oxidative stress was observed in subjects carrying GCPII 1561T and MTRR 66A-variant alleles and low oxidative stress was observed in the subjects carrying cSHMT 1420T and TYMS 5′-UTR 2R allele. GCPII C1561T, MTHFR C677T and MTRR A66G polymorphisms were observed to influence the homocysteine levels (P < 0.05). SHMT and TYMS variants were found to decrease oxidative stress by increasing the folate pool (r = 0.38, P = 0.003) and also by increasing the antioxidant status (r = 0.28, P = 0.03). Influence of dietary folate status was not observed. Overall, this study revealed elevated oxidative stress that was associated with the aberrations in one-carbon metabolism which could possibly influence the CAD risk.     

Plasma Levels of microRNA-145 Are Associated with Severity of Coronary Artery Disease

MethodsA total of 195 consecutive subjects who underwent coronary angiography for chest pain evaluation were enrolled in this study. In CAD patients severity of coronary lesions was assessed by the number of diseased vessels and the Synergy between PCI with Taxus and Cardiac surgery score (SYNTAX score). Plasma levels of miRNA-145 were quantified by real-time quantitative polymerase chain reaction test, and logarithmic transformation of miRNA-145 levels (Ln_miRNA-145) was used for analyses due to its skewed distribution.ResultsOf the 195 total subjects 167 patients were diagnosed as having CAD. Ln_miRNA-145 was significantly lower in CAD patients compared with the non-CAD group (-6.11±0.92 vs. -5.06±1.25; p <0.001). In multivariable linear regression analyses CAD was significantly associated with lower Ln_miRNA-145 (Estimate, -0.50; standard error (SE), 0.11; p <0.0001). Furthermore, among CAD patients, three-vessel disease, higher SYNTAX scores and STEMI were significantly associated with lower Ln_miRNA-145 ([Estimate, -0.40; SE, 0.07; p <0.0001]; [Estimate, -0.02, SE, 0.10; p = 0.005] and [Estimate, -0.35, SE, 0.10; p <0.001] respectively).ConclusionsLower plasma levels of miRNA-145 were significantly associated with the presence as well as severity of CAD. As a potential biomarker for CAD, plasma miRNA-145 may be useful in predicting CAD and its severity in patients presenting with chest pain.     

Variations in ORAI1 Gene Associated with Kawasaki Disease

Kawasaki disease (KD; MIM#61175) is a systemic vasculitis syndrome with unknown etiology which predominantly affects infants and children. Recent findings of susceptibility genes for KD suggest possible involvement of the Ca²⁺/NFAT pathway in the pathogenesis of KD. ORAI1 is a Ca²⁺ release activated Ca²⁺ (CRAC) channel mediating store-operated Ca²⁺ entry (SOCE) on the plasma membrane. The gene for ORAI1 is located in chromosome 12q24 where a positive linkage signal was observed in our previous affected sib-pair study of KD. A common non-synonymous single nucleotide polymorphism located within exon 2 of ORAI1 (rs3741596) was significantly associated with KD (P = 0.028 in the discovery sample set (729 KD cases and 1,315 controls), P = 0.0056 in the replication sample set (1,813 KD cases vs. 1,097 controls) and P = 0.00041 in a meta-analysis by the Mantel-Haenszel method). Interestingly, frequency of the risk allele of rs3741596 is more than 20 times higher in Japanese compared to Europeans. We also found a rare 6 base-pair in-frame insertion variant associated with KD (rs141919534; 2,544 KD cases vs. 2,414 controls, P = 0.012). These data indicate that ORAI1 gene variations are associated with KD and may suggest the potential importance of the Ca²⁺/NFAT pathway in the pathogenesis of this disorder.     

Neutrophil/Lymphocyte Ratio Is Associated with Non-Calcified Plaque Burden in Patients with Coronary Artery Disease

Background

Elevations in soluble markers of inflammation and changes in leukocyte subset distribution are frequently reported in patients with coronary artery disease (CAD). Lately, the neutrophil/lymphocyte ratio has emerged as a potential marker of both CAD severity and cardiovascular prognosis.

Objectives

The aim of the study was to investigate whether neutrophil/lymphocyte ratio and other immune-inflammatory markers were related to plaque burden, as assessed by coronary computed tomography angiography (CCTA), in patients with CAD.

Methods

Twenty patients with non-ST-elevation acute coronary syndrome (NSTE-ACS) and 30 patients with stable angina (SA) underwent CCTA at two occasions, immediately prior to coronary angiography and after three months. Atherosclerotic plaques were classified as calcified, mixed and non-calcified. Blood samples were drawn at both occasions. Leukocyte subsets were analyzed by white blood cell differential counts and flow cytometry. Levels of C-reactive protein (CRP) and interleukin(IL)-6 were measured in plasma. Blood analyses were also performed in 37 healthy controls.

Results

Plaque variables did not change over 3 months, total plaque burden being similar in NSTE-ACS and SA. However, non-calcified/total plaque ratio was higher in NSTE-ACS, 0.25(0.09–0.44) vs 0.11(0.00–0.25), p<0.05. At admission, levels of monocytes, neutrophils, neutrophil/lymphocyte ratios, CD4+ T cells, CRP and IL-6 were significantly elevated, while levels of NK cells were reduced, in both patient groups as compared to controls. After 3 months, levels of monocytes, neutrophils, neutrophil/lymphocyte ratios and CD4+ T cells remained elevated in patients. Neutrophil/lymphocyte ratios and neutrophil counts correlated significantly with numbers of non-calcified plaques and also with non-calcified/total plaque ratio (r = 0.403, p = 0.010 and r = 0.382, p = 0.024, respectively), but not with total plaque burden.

Conclusions

Among immune-inflammatory markers in NSTE-ACS and SA patients, neutrophil counts and neutrophil/lymphocyte ratios were significantly correlated with non-calcified plaques. Data suggest that these easily measured biomarkers reflect the burden of vulnerable plaques in CAD.     

Pericardial Fat Is Associated with Coronary Artery Calcification in Non-Dialysis Dependent Chronic Kidney Disease Patients

Pericardial fat (PF) a component of visceral adipose tissue has been consistently related to coronary atherosclerosis in the general population. This study evaluated the association between PF and coronary artery calcification (CAC) in non-dialysis dependent chronic kidney disease (CKD) patients. This is a post-hoc cross sectional analysis of the baseline of a prospective cohort of 117 outward CKD patients without manifest coronary artery disease (age, 56.9±11.0 years, 64.1% males, 95.1% hypertensives, 25.2% diabetics, 15.5% ever smokers, CKD stage 2 to 5 with estimated glomerular filtration rate 36.8±18.1 ml/min). CAC scores, PF volume and abdominal visceral fat (AVF) areas were measured by computed tomography. The association of PF as a continuous variable with the presence of CAC was analyzed by multivariate logistic regression. CAC (calcium score >0) was present in 59.2% patients. Those presenting CAC were on average 10 years older, had a higher proportion of male gender (78.7% vs. 42.9%, p<0.001), and had higher values of waist circumference (95.9±10.7 vs. 90.2±13.2 cm, p = 0.02), PF volumes (224.8±107.6 vs. 139.1±85.0 cm³, p<0.01) and AVF areas (109.2±81.5 vs. 70.2±62.9 cm², p = 0.01). In the multivariate analysis, adjusting for age, gender, diabetes, smoking and, left ventricular concentric hypertrophy, PF was significantly associated with the presence of CAC (OR: 1.88 95% CI: 1.03–3.43 per standard deviation). PF remained associated with CAC even with additional adjustments for estimated glomerular filtration rate or serum phosphorus (OR: 1.85 95% CI: 1.00–3.42, p = 0.05). PF is independently associated with CAC in non-dialysis dependent CKD patients.