Oxidised Low-Density Lipoprotein and Its Receptor-Mediated Endothelial Dysfunction Are Associated with Coronary Artery Lesions in Kawasaki Disease.

The study aimed to investigate the role of oxidised low-density lipoprotein (oxLDL)/lectin-like-oxLDL receptor-1 (LOX-1) in coronary artery lesions (CALs) in Kawasaki disease (KD) and of plasma oxLDL concentration in the early prediction of CALs in KD. This prospective study included 80 KD patients, 20 febrile and 20 healthy children. oxLDL, LOX-1 and other parameters were analysed in the acute phase. Plasma oxLDL concentration and LOX-1 mRNA expression in peripheral blood mononuclear cells (PBMCs) were significantly increased in KD patients compared with febrile and healthy children (P < 0.001 and P = 0.022, respectively), particularly in the group with CALs (P < 0.001 and P = 0.027, respectively). Coronary Z-score was significantly correlated with plasma oxLDL concentration and LOX-1 mRNA expression (r = 0.739 and 0.637, respectively; P < 0.01). The sensitivity and specificity of predicting CALs were 71.4% and 77.2%, respectively, at plasma oxLDL concentration ≥ 12.38 mU/L. oxLDL/LOX-1 may be involved in CAL development. The plasma oxLDL concentration in the acute phase is a potentially useful biological indicator for predicting CAL in KD patients.

Microarray analysis reveals key genes and pathways in Tetralogy of Fallot.

The aim of the present study was to identify key genes that may be involved in the pathogenesis of Tetralogy of Fallot (TOF) using bioinformatics methods. The GSE26125 microarray dataset, which includes cardiovascular tissue samples derived from 16 children with TOF and five healthy age­matched control infants, was downloaded from the Gene Expression Omnibus database. Differential expression analysis was performed between TOF and control samples to identify differentially expressed genes (DEGs) using Student's t­test, and the R/limma package, with a log2 fold­change of >2 and a false discovery rate of <0.01 set as thresholds. The biological functions of DEGs were analyzed using the ToppGene database. The ReactomeFIViz application was used to construct functional interaction (FI) networks, and the genes in each module were subjected to pathway enrichment analysis. The iRegulon plugin was used to identify transcription factors predicted to regulate the DEGs in the FI network, and the gene­transcription factor pairs were then visualized using Cytoscape software. A total of 878 DEGs were identified, including 848 upregulated genes and 30 downregulated genes. The gene FI network contained seven function modules, which were all comprised of upregulated genes. Genes enriched in Module 1 were enriched in the following three neurological disorder­associated signaling pathways: Parkinson's disease, Alzheimer's disease and Huntington's disease. Genes in Modules 0, 3 and 5 were dominantly enriched in pathways associated with ribosomes and protein translation. The Xbox binding protein 1 transcription factor was demonstrated to be involved in the regulation of genes encoding the subunits of cytoplasmic and mitochondrial ribosomes, as well as genes involved in neurodegenerative disorders. Therefore, dysfunction of genes involved in signaling pathways associated with neurodegenerative disorders, ribosome function and protein translation may contribute to the pathogenesis of TOF.

[Effects of carvedilol on cardiomyocyte apoptosis in autoimmune myocarditis in mice].

OBJECTIVE: To observe the effects of carvedilol on the expression of Bcl-2, Bax and Fas in autoimmune myocarditis (AM). METHODS: A total of 60 inbred male BALB/C mice 4 - 5 weeks of age were divided at random into 3 groups as follows: AM group (n = 20), carvedilol group (n = 20) and control group (n = 20). The mice were sacrificed after gathering blood specimens by taking out the eyeballs and hearts tissue. The histological and ultrastructural changes were observed under light microscope and electron microscope. The concentrations of cardiac troponin I (cTn I) were detected by chemiluminescence immunoassay (CLIA). Immunohistochemistry (IHC) was performed to analyze the contents of Bcl-2, Bax and Fas, TUNEL to detect the apoptotic index in myocardial cells. RESULTS: There were large number of lymphocyte and monocyte infiltrates under light microscope and karyopyknosis and chromatin gathered along the nuclear membrane under electron microscope in AM group. There were no inflammations and chromatin gathering in group C. Compared with control group, the Bcl-2, Bax and Fas protein expression significantly elevated in AM group (23.48 ± 2.24 vs. 6.64 ± 1.60, 26.15 ± 2.02 vs. 5.09 ± 0.85, 21.22 ± 3.62 vs. 5.86 ± 1.37, P < 0.01). The histopathologic scores (2.60 ± 0.31 vs. 2.02 ± 0.26, P < 0.05) and karyopyknosis of carvedilol group decreased as compared with AM group. The Bcl-2, Bax and Fas protein expression (17.13 ± 1.94 vs. 23.48 ± 2.24, 17.66 ± 2.62 vs. 26.15 ± 2.02, 16.79 ± 2.83 vs. 21.22 ± 3.62, P < 0.05), AI [(16.61 ± 4.67)% vs. (24.51 ± 4.70)%, P < 0.05] and contents of cTnI [(1.878 ± 0.48) ng/ml vs. (1.102 ± 0.23) ng/ml, P < 0.05] also decreased in carvedilol group compared with AM group. CONCLUSION: Carvedilol could protect against AM by alleviating cardiomyocyte apoptosis.

[Implication of N-terminal pro-B-type natriuretic peptide and Tei index of left ventricle changes in children with ventricular septal defect treated by transcatheter interventional therapy].

OBJECTIVE: To explore the implication of the dynamic changes of plasma N-terminal pro-B-type natriuretic peptide (NT-proBNP) level and Tei index of left ventricle (LV) in children with ventricular septal defect (VSD) treated by transcatheter closure. METHODS: Sixty children with VSD treated by transcatheter closure with VSD occluder (Group VSD) and 30 healthy children (Group C) were included in this study. The plasma concentration of NT-proBNP, Tei index of LV and left ventricle ejection fraction (LVEF) were measured in Group C and at before, 5th minute, 4th hour, 1st month, 3rd month and 6th month after VSD closure in Group VSD. RESULTS: (1) The concentration of plasma NT-proBNP was significantly increased in children with VSD before transcatheter closure compared with Group C [(229.45 ± 57.75) ng/L vs. (99.21 ± 46.86) ng/L, P < 0.01], significantly increased at 5th minute and 24th hour after transcatheter closure [(356.27 ± 96.78) ng/L and (356.38 ± 91.95) ng/L vs. (229.45 ± 57.75) ng/L, all P < 0.01], and significantly decreased at 1st month, 3rd months and 6th months after transcatheter closure [(131.33 ± 34.79) ng/L, (96.56 ± 31.55) ng/L and (93.39 ± 29.46) ng/L vs. (229.45 ± 57.75) ng/L, P < 0.05 or P < 0.01]. (2) The Tei indexes of LV in Group VSD before transcatheter closure were significantly higher than in Group C (0.45 ± 0.05 vs. 0.33 ± 0.08, P < 0.01) and Tei index was significantly increased at 24th hour, 1st month after transcatheter closure (P < 0.01) while significantly decreased at 3rd and 6th month compared with those before transcatheter closure (0.34 ± 0.07 and 0.34 ± 0.06 vs. 0.45 ± 0.05, all P < 0.01). (3) There is a positive correlation between the changes of the plasma concentration of NT-proBNP and the change of Tei index of LV before and after transcatheter closure (r = 0.653, P < 0.05). CONCLUSION: Tei index of LV and NT-proBNP can monitor cardiac function changes in children with VSD before and after transcatheter closure.

[Heart rate variability, adrenomedullin and B-type natriuretic peptide before and after transcatheter closure in children with patent ductus arteriosus].

OBJECTIVE: To observe the changes of heart rate variability (HRV), adrenomedullin (ADM) and B-type natriuretic peptide (BNP) before and after transcatheter closure in children with patent ductus arteriosus. METHODS: HRV spectral values (TF, VLF, LF, HF, LF/HF) were detected by 24 dynamic electrocardiogram and the concentrations of plasma ADM and BNP were measured in 55 children with patent ductus arteriosus (Group PDA, n = 55) before and 3(rd) day, 3(rd) month after transcatheter closure therapy, and in 60 normal children (Group C). RESULTS: (1) Compared with Group C, the HRV spectral values (TF, VLF, HF) were significantly lower (all P < 0.01), LF/HF and the concentrations of plasma ADM, BNP were significantly higher in patients with PDA before transcatheter closure (all P < 0.01). (2) Compared with the values before transcatheter closure values, plasma ADM were significantly reduced at 3(rd) day and 3(rd) month after transcatheter closure (P < 0.01), the HRV spectral values (TF, VLF, HF) were significantly increased while LF/HF and plasma BNP were significantly decreased at 3(rd) month after transcatheter closure (P < 0.01 or P < 0.05). CONCLUSIONS: HRV and plasma ADM, BNP improved significantly post transcatheter closure in children with patent ductus arteriosus.

[Changes of serum leptin and vascular endothelial growth factor in children with congenital heart disease].

OBJECTIVE: To study the changes of serum leptin (LEP) and vascular endothelial growth factor (VEGF) in children with congenital heart disease(CHD) and their roles in CHD. METHODS: Forty-eight children with acyanotic CHD (ACHD group), 20 age-matched children with cyanotic CHD (CCHD group) and 20 healthy children (control group) were enrolled. The ACHD group was subdivided into two groups with (n=20) or without concurrent heart failure (n=28). Serum LEP, VEGF, total protein and albumin levels and body mass index (BMI) were measured. RESULTS: Serum total protein and albumin levels were not apparently different in all CHD children from healthy controls, but there was a significant difference in the BMI between them (p<0.01). Serum LEP and VEGF levels and the ratio of LEP/BMI in all CHD children were significantly higher than those in healthy controls (p<0.01). Compared with the ACHD group without heart failure, the serum LEP and VEGF levels and the ratio of LEP/BMI in the CCHD and the ACHD with heart failure groups increased significantly (p<0.01). In the ACHD group, serum LEP level was positively correlated with BMI (p<0.01). In the CCHD group, there were positive correlations between serum LEP level and serum VEGF level (p<0.01) and between hemoglobin concentration and serum VEGF level (p<0.01). Arterial oxygen saturation was negatively correlated with serum VEGF (p<0.01) and LEP levels (p<0.01) in the CCHD group. CONCLUSIONS: Both VEGF and LEP play roles in the pathophisiological process of CHD. VEGF and LEP are associated with the development of heart failure in children with ACHD.

[Reducing plasma homocysteic acid lowers serum C-reactive protein level in children with Kawasaki disease].

OBJECTIVE: To investigate the effect of plasma homocysteic acid (HCA) reduction on serum C-reactive protein (CRP) level in children with Kawasaki disease (KD). METHODS: Seventy-six children with KD were divided into 2 equal groups for treatment with aspirin and IVIG, or with vitamin B6 and folic acid besides in addition to aspirin and IVIG. Serum CRP level was tested before and after the treatments, and plasma HCA level was also measured after the treatments. RESULTS: Serum CRP level was comparable between the two groups before the treatment, but significantly reduced after vitamin B6 and folic acid treatment (7.56-/+2.94 mg/L vs 12.23-/+4.16 mg/L, P<0.05). Additional vitamin B6 and folic acid treatment significantly lowered plasma HCA level (4.56-/+1.14 micromol/L vs 7.79-/+1.79 micromol/L, P<0.05), and correlation analysis demonstrated a positive correlation between plasma HCA and serum CRP levels (r=0.697, P<0.01). CONCLUSION: Lowering plasma HCA can decrease serum CRP in children with KD to enhance the therapeutic effect.

[Effects of hemoglobin oxygenase-1 inhibitor zinc protoporphyrin IX on acute viral myocarditis in mice].

OBJECTIVE: To investigate the role of heme oxygenase-1 (HO-1) and its catalyst carbon monoxide (CO) in the development of myocardial damage and the effects of zinc protoporphyrin IX (ZnPPIX), an inhibitor of HO-1 on myocardium of mice with acute viral myocarditis. METHODS: A total of 112 inbred male Balb/C mice 4 - 6 weeks of age were divided randomly into 3 groups: the control group (C group, n = 32), the viral myocarditis group (V group, n = 40) and ZnPPIX group (Z group, n = 40). The Z and V groups were inoculated intraperitoneally (i.p.) with 0.1 ml of 10(-4.36) tissue culture infectious dose 50% (TCID(50))/ml Coxsackie virus B3 (CVB(3)) to produce viral myocarditis model on day 0, C group was injected i.p. with virus-free 1640 culture culture medium 0.1 ml at the same time, then operation was done as follows: the mice of group C and group V were injected i.p. with 0.1 ml NS each day. The mice of group Z were injected i.p. with 40 micromol per kilogram of body weight ZnPPIX (HO-1 inhibitor) qod. Eight mice of each group were sacrificed on days 4, 8, 15 and 21, respectively. The blood specimens were collected by taking out the eyeballs to test for the content of carboxyhemoglobin (COHb) using spectrophotometry and cardiac troponin I (cTnI) using chemiluminescent immunoassay. The hearts tissue slides were also stained by immunohistochemistry (IHC) for HO-1 and in situ hybridization (ISH) for HO-1 mRNA. The histological and ultrastructural changes were observed under light and electron microscopes. RESULTS: (1) The histopathological changes of myocardial cells: in the V and Z groups myocardial inflammatory cells infiltration reached the peak on day 8, the Z group histopathological scores were significantly lower than those in V group on day 8 (2.40 +/- 0.31 vs. 1.73 +/- 0.24, P < 0.01) and on day 15 (1.78 +/- 0.29 vs. 1.43 +/- 0.23, P < 0.05). No inflammation was present in group C. (2) The changes of serum cTnI level in both V and Z groups were significantly higher than those in C group on day 4, 8 and 15 (P < 0.01). The level in Z group was significantly lower than that in V group on day 4 [(6.074 +/- 1.475) ng/ml vs (7.911 +/- 1.225) ng/ml, P < 0.05] and day 8 [(0.821 +/- 0.294) ng/ml vs (1.480 +/- 0.454) ng/ml, P < 0.05]. (3) The changes of blood COHb level: compared with V group, in Z group the COHb level was lower on day 4 (P < 0.05) and day 15 (P < 0.01) after CVB(3) inoculation. Surprisingly, in Z group COHb level elevated suddenly on day 8 and showed conspicuously higher than that of V group (P < 0.01). (4) The result of HO-1 IHC staining: in both V and Z group myocardial cells had positive expression, while C group did not. (5) The results of HO-1 ISH were similar to those of HO-1 IHC, the A values of group Z was significantly lower than that of group V on day 4, 15 and 21(P < 0.01), but on day 8 it was higher than that of group C (P < 0.05). CONCLUSION: HO-1 inhibitor, ZnPP not only could inhibit HO-1 overexpression but also could induce HO-1 expression temporarily and protect against myocardial injury at the early stage of acute viral myocarditis.

[Expression and pathological implication of transforming growth factor-beta1 mRNA and endothelin-1 mRNA in intraacinar pulmonary arterioles of congenital heart disease accompanied with pulmonary hypertension].

OBJECTIVE: To study the expression and pathological implication of transforming growth factor-1 (TGF-1) and endothelin-1 (ET-1) in intraacinar pulmonary arterioles of children with congenital heart disease and pulmonary hypertension (HP). METHODS: Forty-one children with left-to-right shunt congenital heart disease were studied including 25 cases of HP (group A), 16 cases without HP (group B) and 10 children without congenital heart disease as the contols (group C). Expression of TGF-beta1 mRNA and ET-1 mRNA in intraacinar pulmonary arteriolar (IAPA) was studied using in-situ hybridization and image pattern analysis of their absorption values (A value). Changes of the intraacinar arterioles and lung tissue were studied by elastic fiber staining and electronic microscopy respectively. RESULTS: (1) There was a significant difference in the amount of intraacinar pulmonary arterioles (partial-muscular and muscular) counted in either group A or B in comparing with that of group C (F values 149.96 and 142.01 respectively, P < 0.01); (2) Electronic microscopy demonstrated endothelial proliferation of the small arteries, thickening of arteriolar wall, increased density of collagen fibers at adventitia and increased thickness of the capillary basal membrane; (3) The A value of TGF-beta1 mRNA expressed in the pulmonary arterioles of groups A and B by in-situ hybridization were 0.1988 +/- 0.0498 and 0.1098 +/- 0.0428 respectively, however, the expression was weak in group C (A value: 0.0578 +/- 0.0096). There were all significant between each two groups (F = 45.95, P < 0.01). The expression of ET-1 mRNA was markedly increased as well in the endothelial cells of pulmonary arterioles in both groups A and B, with A values of 0.1692 +/- 0.0205 and 0.1004 +/- 0.0140 respectively, whereas the expression was weak in group C (A value of 0.0746 +/- 0.0119). There were all significant between each two groups (F = 139.996, P < 0.01). CONCLUSIONS: The number of intraacinar pulmonary partial-muscular and muscular arterioles in patients with left-to-right shunt congenital heart defect is drastically increased, along with marked restructuring of the pulmonary vasculatures. In addition, there seems a correlation present between the overexpression of TGF-beta1 mRNA and ET-1 mRNA in intraacinar pulmonary arterioles and the occurrence of pulmonary hypertension in patients with congenital heart disease.