Clinicopathological and genetic characteristics of gastric neuroendocrine tumour (NET) G3 and comparisons with neuroendocrine carcinoma and NET G2.

AIMS: To characterise the clinicopathological and genetic characteristics of gastric neuroendocrine tumour G3 (gNET G3) and to compare them with those of gastric neuroendocrine carcinoma (gNEC) and gNET G2. METHODS AND RESULTS: A total of 115 gastric neuroendocrine neoplasms (NENs) were included, of which gNET G3 was different from gNET G1/G2 in terms of tumour location (P = 0.029), number (P = 0.003), size (P = 0.010), the Ki67 index (P < 0.001), lymph node metastasis (P < 0.001) and TNM stage (P = 0.011), and different from gNEC/gastric mixed neuroendocrine-non-neuroendocrine neoplasm (gMiNEN) in terms of tumour size (P = 0.010) and the Ki67 index (P = 0.001). High-resolution copy number (CN) profiling and validation experiments showed CN gains and high expression of DLL3 in gNET G3. Hierarchical clustering analysis based on CN characteristics showed that gNET G3 was separated from gNEC but mixed with gNET G2. In gene set enrichment analysis, eight pathways were significantly enriched in gNEC when comparing gNET G3 and gNEC (P < 0.05), while no pathways were enriched when comparing gNET G3 and gNET G2. Whole-exome sequencing and validation experiments showed nonsense mutation of TP53 in one gNET G3, with wild-type staining for p53. In gNEC, TP53 mutations were detected in four of eight cases, and abnormal expression of p53 was detected in all cases. CONCLUSION: Gastric NET G3 is a distinct entity with unique genetic characteristics, which are different from those of gNEC than gNET G2. Our results provide insight into some molecular alterations that may contribute to the development and progression of gNET G3 and serve as potential therapeutic targets.

Cleavage of IκBα by calpain induces myocardial NF-κB activation, TNF-α expression, and cardiac dysfunction in septic mice.

Recent studies in septic models have shown that myocardial calpain activity and TNF-α expression increase during sepsis and that inhibition of calpain activation downregulates myocardial TNF-α expression and improves cardiac dysfunction. However, the mechanism underlying this pathological process is unclear. Thus, in the present study, we aimed to explore whether IκBα/NF-κB signaling linked myocardial calpain activity and TNF-α expression in septic mice. Adult male mice were injected with LPS (4 mg/kg ip) to induce sepsis. Myocardial calpain activity, IκBα/NF-κB signaling activity, and TNF-α expression were assessed, and myocardial function was evaluated using the Langendorff system. In septic mice, myocardial calpain activity and TNF-α expression were increased and IκBα protein was degraded. Furthermore, NF-κB was activated, as indicated by increased NF-κB p65 phosphorylation, cleavage of p105 into p50, and its nuclear translocation. Administration of the calpain inhibitors calpain inhibitor Ш and PD-150606 prevented the LPS-induced degradation of myocardial IκBα, NF-κB activation, and TNF-α expression and ultimately improved myocardial function. In calpastatin transgenic mice, an endogenous calpain inhibitor and cultured neonatal mouse cardiomyocytes overexpressing calpastatin also inhibited calpain activity, IκBα protein degradation, and NF-κB activation after LPS treatment. In conclusion, myocardial calpain activity was increased in septic mice. Calpain induced myocardial NF-κB activation, TNF-α expression, and myocardial dysfunction in septic mice through IκBα protein cleavage.

Heat shock transcription factor-1 inhibits H2O2-induced apoptosis via down-regulation of reactive oxygen species in cardiac myocytes.

Heat shock transcription factor-1 (HSF1) protects against cardiac diseases such as ischemia/reperfusion injury and myocardial infarction. However, the mechanisms have not yet been fully characterized. In this study, we investigated the effects of reactive oxygen species (ROS) and apoptosis signal-regulating kinase-1 (ASK1) in HSF1-regulated cardiomyocyte protection. Cultured cardiomyocytes of neonatal rats were transfected with HSF1, ASK1 or both of them before exposure to H(2)O(2), and the ROS generation, c-Jun N-terminal kinase (JNK) activity and apoptosis were examined. H(2)O(2) significantly increased intracellular ROS generation and apoptotic cells as expected, and all these cellular events were greatly inhibited by overexpression of HSF1. However, H(2)O(2)-induced increases in JNK phosphorylation and cell apoptosis were largely enhanced by ASK1 overexpression whereas the similar transfection did not affect the ROS generation in the cells. Moreover, inhibition of H(2)O(2)-increased ROS generation, JNK phosphorylation, and cellular apoptosis by overexpression of HSF1 tended to be disappeared, when the cells were co-transfected with ASK1. These results suggest that HSF1 protects cardiomyocytes from apoptosis under oxidative stress via down-regulation of intracellular ROS generation and inhibition of JNK phosphorylation. Although ASK1 itself has no effect on intracellular ROS generation, it may affect the inhibitory effects of HSF1 on ROS generation, JNK activity, and cardiomyocyte injury.

A modified formula for calculating low-density lipoprotein cholesterol values.

BACKGROUND: The Friedewald formula (FF) is useful for calculating serum low-density lipoprotein cholesterol (LDL-C) values, but has a remarkable deviation and limitation especially in hypertriglyceridemia. We modify the formula which is now more suitable for LDL-C calculation. METHODS: 2180 cases were classified into three groups according to their TG concentrations (A: < 200 mg/dl, n = 1220; B: 200-400 mg/dl, n = 480; C: 400-1000 mg/dl, n = 480). The concentrations of LDL-C were measured or estimated by 1) a direct measurement (DM); 2) the FF; and 3) our modified Friedewald formula (MFF): LDL-C (mg/dl) = Non-HDL-C x 90% - TG x 10%. RESULTS: Linear regression showed a significant correlation (P < 0.001) between the measured and calculated LDL-C values. Bland-Altman plots indicated that the methods (DM/MFF) were in better agreement than those (DM/FF). The LDL-C/Non-HDL-C ratio in FF calculated values was significantly lower (P < 0.05) than that in MFF or DM values, while no significant difference between MFF and DM was found. In Group A and Group B, 4.26% and 14.79% of the MFF calculated values had more than 20% deviation from those measured by DM. These percentages were significantly lower than those calculated by FF, where 7.30% and 25.63% were observed, respectively (P < 0.01 and P < 0.001). The MFF calculated values were all positive even in Group C. CONCLUSIONS: Compared with the FF calculation, serum LDL-C values estimated by our modified formula are closer to those measured by a direct assay. The modification significantly diminishes the interference caused by hypertriglyceridemia.

[Association between myocardial calpain activation and apoptosis in lipopolysaccharide-induced septic mouse model].

OBJECTIVE: in septic mice, myocardial calpain was activated and induced caspase-3 activation, the association between calpain activation and apoptosis was explored in this experiment. METHODS: in in vivo model, adult C57 mice were injected with lipopolysaccharide (LPS, 4 mg/kg, i.p.) to induce sepsis. Myocardial calpain and caspase-3 activities, protein levels of calpain-1, calpain-2, calpastatin, Bcl-2 and Bid were detected by Western blot analysis and myocardial apoptosis was detected by TUNEL, myocardiac function was evaluated by Langendorff system. In in vitro model, adult rat cardiomyocytes were incubated with LPS (1 microg/ml) or co-incubated with calpain inhibitor-III (10 micromol/L), calpain activity, caspase-3 activity, protein levels of Bcl-2 and Bid, and cardiomyocyte apoptosis were detected. RESULTS: in septic mice, myocardial calpain and caspase-3 activity were increased up to 2.7- and 1.8-folds, respectively. Both calpain inhibitor-III and PD150606 significantly attenuated the increase of caspase-3 activity. Myocardial protein levels of calpain-1, calpain-2, calpastatin, Bcl-2 and Bid were similar between control and septic mice, and no cleavage of both Bcl-2 and Bid was found in septic mice. Calpain inhibitor-III significantly improved myocardial function in septic mice. In in vitro model, calpain and caspase-3 activities were increased after 4 h LPS treatment, co-treatment with calpain inhibitor-III prevented caspase-3 activity increase, protein Bcl-2 and Bid were similar between normal cardiomyocytes and LPS-treated cardiomyocytes. Cardiomyocyte apoptosis was similar in in vivo and in vitro septic models. CONCLUSION: myocardial calpain activity is increased in LPS induced septic mice, subsequent caspase-3 activation may contribute to myocardial dysfunction in septic mice without aggravating myocardial apoptosis and Bcl-2 and Bid are not involved on calpain induced caspase-3 activation in our model.

[Overexpression of endothelin-1 induces hypertrophy of rat pulmonary arterial microvascular smooth muscle cells via Akt/mTOR pathway].

OBJECTIVE: To investigate the effects of endothelin-1 (ET-1) overexpression on hypertrophy of the rat pulmonary arterial microvascular smooth muscle cells (RPMCs) in vitro. METHODS: Lung tissue perfusion method was used to obtain the primary RPMCs, and then the cells were transiently transfected with ET-1 plasmids or empty vectors via Lipofectamine. Flow cytometry was used to determine the distributions of the cell cycle and cell size of RPMCs. Immunofluorescence staining and Western blotting were used to determine the level of alpha-smooth muscle actin. The ratio of the protein/DNA of the transfected RPMCs was also calculated. Western blotting was used to examine the levels of phosphorylations of protein kinase B/Akt, mTOR, and ERK1/2. RESULTS: Primary RPMCs were isolated successfully. Overexpression of ET-1 resulted in a significant increase in total protein synthesis, expression of alpha-SMA, as well as increased cell size. Western blotting results showed that overexpression of ET-1 resulted in increased phosphorylation of Akt, mTOR and a decreased phosphorylation of ERK1/2 in RPMC. CONCLUSIONS: ET-1 overexpression induces RPMC hypertrophy and activation of Akt/PKB-mTOR pathway may be involved in the mechanism, with important implications for the pathogenesis of vascular remodeling in pulmonary arterial hypertension.

[Link between cardiac myosin binding protein-C gene mutation of Pro1208fs and Gly507 Arg and hypertrophic cardiomyopathy in Chinese patients].

OBJECTIVE: To detect gene mutations associated with hypertrophic cardiomyopathy (HCM) in Chinese patients and possible correlations between genotype and phenotype. METHODS: Twenty-one unrelated patients with hypertrophic cardiomyopathy were studied. The clinical data including symptoms, physical examination, echocardiography and electrocardiography were collected. The full ecoding exons of cardiac myosin-binding protein C gene (cMYBPC3) were amplified with PCR and the products were sequenced. RESULTS: Two mutations were identified in probands from two families. One mutation was frame shift mutation Pro1208fs in the exon 32 of the cMYBPC3 gene. Pro1208fs mutation was identified in a 59 years old female patient with familial hypertrophic cardiomyopathy. Symptom onset was late and a favorable clinical course was evidenced in this patient. Another mutation was missence mutation Gly507Arg in the exon 17 of the MYBPC3 gene identified in a 24 years old male patient. Diffuse thickness of left ventricular wall, impaired diastolic function and enlarged left atria were evidenced in echocardiography. No mutation was identified in the 80 control healthy individuals. CONCLUSION: cMYBPC3 might be the disease-causing genes in Chinese patients with hypertrophic cardiomyopathy.

Evacetrapib reduces preß-1 HDL in patients with atherosclerotic cardiovascular disease or diabetes.

BACKGROUND AND AIMS: Cholesteryl ester transfer protein (CETP) inhibitor-mediated induction of HDL-cholesterol has no effect on the protection from cardiovascular disease (CVD). However, the mechanism is still unknown. Data on the effects of this class of drugs on subclasses of HDL are either limited or insufficient. In this study, we investigated the effect of evacetrapib, a CETP inhibitor, on subclasses of HDL in patients with atherosclerotic cardiovascular disease or diabetes. METHODS: Baseline and 3-month post-treatment samples from atorvastatin 40 mg plus evacetrapib 130 mg (n = 70) and atorvastatin 40 mg plus placebo (n = 30) arms were used for this purpose. Four subclasses of HDL (large HDL, medium HDL, small HDL, and preß-1 HDL) were separated according to their size and quantified by densitometry using a recently developed native polyacrylamide gel electrophoresis (PAGE) system. RESULTS: Relative to placebo, while evacetrapib treatment dramatically increased large HDL and medium HDL subclasses, it significantly reduced small HDL (27%) as well as preß-1 HDL (36%) particles. Evacetrapib treatment reduced total LDL, but also resulted in polydisperse LDL with LDL particles larger and smaller than the LDL subclasses of the placebo group. CONCLUSION: Evacetrapib reduced preß-1 HDL and small HDL in patients with ASCVD or diabetes on statin. Preß-1 HDL and medium HDL are negatively interrelated. The results could give a clue to understand the effect of CETP inhibitors on cardiovascular outcomes.

[Endothelin-1 overexpression inhibits rat pulmonary arterial microvascular smooth muscle cell apoptosis via Akt/PKB pathway].

OBJECTIVE: To investigate the effects of endothelin-1 (ET-1) overexpression on apoptosis of the rat pulmonary arterial microvascular smooth muscle cells (RPMC) in vitro. METHODS: Primary RPMC obtained from the pulmonary artery and lung microvasculature were identified by immunofluorescence staining and electron microscope technique. The RPMC was transient transfected with the pMEXneo-ET1 and pCDNA5-FRT-TO-ET1-3'UTR plasmids as well as the empty vector respectively via lipofectamine. Flow cytometry was used to assess the cell cycle and apoptosis of RPMC. Akt and Caspase-3 expressions were detected by Western blot and real time RT-PCR. RESULTS: The mRNA of ET(A) expression was significantly higher than that of ET(B) receptor in primary RPMC. Flow cytometry analysis revealed significantly reduced apoptosis in ET-1 transfected RPMC compared to that in vehicle transfected RPMC. Overexpression of ET-1 in RPMC also significantly increased the phosphorylation of Akt and reduced the cleaved Caspase-3 expression. CONCLUSIONS: Overexpression of the ET-1 inhibited RPMC apoptosis and activated Akt/PKB-Caspase-3 signaling pathway, which might be responsible for ET-1 induced the pulmonary microvascular arteries remodeling.

Endothelial­to­mesenchymal transition in human idiopathic dilated cardiomyopathy.

Dilated cardiomyopathy (DCM) is characterized by left ventricular dilation and cardiac fibrosis. Emerging evidence indicated that endothelial­to­mesenchymal transition (Endo­MT) is a crucial event during organ fibrosis. This study was performed to clarify whether Endo­MT contributed to the progression of cardiac fibrosis in DCM. Cardiac samples from patients with DCM and control were obtained. The presence of endothelial markers, cluster of differentiation (CD)31 and vascular endothelial (VE)­cadherin, and mesenchymal markers, α smooth muscle actin (SMA) and fibroblast­specific protein 1 (FSP1) was performed using immunohistochemistry. Co­localization of endothelial markers and mesenchymal markers were identified using confocal immunofluorescence staining. Serum procollagen type I carboxy­terminal propeptide (PICP) and procollagen type III amino­terminal propeptide (PIIINP) were measured by ELISA. Protein levels of Wnt, ß­catenin and Snail were determined using western blot analysis. Immunohistochemistry and double­immunofluorescence staining demonstrated that the expression of CD31 and VE­cadherin were significantly decreased in DCM samples, whereas the FSP­1, and αSMA were significantly increased. CD31 and VE­cadherin labeling indexes were respectively negatively correlated with left ventricular end­diastolic diameter (LVEDD) (CD31 r=­0.82, P<0.01; VE­cadherin r=-0.73, P<0.01), while FSP­1 and αSMA were positively associated with LVEDD (αSMA r=0.65, P<0.01, FSP1 r=0.53, P<0.01) and left ventricular ejection fraction (αSMA r=­0.18, P<0.05; FSP1 r=­0.21, P<0.05). Furthermore, PICP and PIIINP levels were positively associated with the co­expression labeling indexes (CD31/SMA co­labeling index and PICP r=0.727, P<0.01; CD31/SMA co­labeling index and PIIINP r=0.741, P<0.01; VE­Cadherin/FSP­1 co­labeling index and PICP r=0.716, P<0.01; VE­cadherin/FSP­1 co­labeling index and PIIINP r=0.648, P<0.05). Western blot analysis indicated that proteins levels of Wnt signaling and snail were significantly increased in DCM samples. These results suggested that Endo­MT is potentially implicated in the pathogenesis of myocardial fibrosis and remodeling during the development of DCM, indicating a potential therapeutic target for DCM treatment.

Real-time myocardial contrast echocardiography can predict functional recovery and left ventricular remodeling after revascularization in patients with ischemic heart disease.

BACKGROUND: Previous studies showed that preservation of microvascular integrity after myocardial ischemia was associated with myocardial viability. Real-time myocardial contrast echocardiography (RT-MCE) is a promising modality for non-invasive evaluation of microcirculation perfusion. Thus, it provides a unique tool to detect myocardial viability. We sought in this study to investigate the role of RT-MCE in predicting left ventricular (LV) functional recovery and remodeling after revascularization in patients with ischemic heart disease. METHODS: Thirty-one patients with ischemic heart disease and resting regional LV dysfunction were included. LV volume, global and regional function were evaluated by echocardiography before and 6 - 9 months after revascularization. RT-MCE was performed before revascularization using low mechanical index power modulation imaging. Myocardial contrast opacification of dysfunctional segments was scored on a 3-point scale and mean contrast score in dysfunctional segments was calculated. Patients were divided into 2 groups according to mean contrast score in dysfunctional segments: group A, patients with mean contrast score = 0.5 (n = 19); group B, patients with mean contrast score < 0.5 (n = 12). RESULTS: Wall motion improvement was found to be 94.5%, 45.5% and 16.1% respectively (P < 0.01) in homogenous, patchy and absent contrast opacification segments. At baseline, there was no significant difference in LV volume and global function between the two groups. After revascularization, group B had significantly larger LV end-diastolic volume (LVEDV) and LV end-systolic volume (LVESV), lower LV ejection fraction (LVEF) and higher wall motion score index (WMSI) than those of group A (all P < 0.05). Revascularization was followed by significant improvement of LV volume and recovery of global LV function in group A (all P < 0.01); however, in group B, after revascularization, deterioration of LVEDV (P < 0.05) was observed, moreover LVESV, WMSI and LVEF did not change significantly. CONCLUSIONS: The maintenance of myocardial microcirculation detected by RT-MCE can predict functional recovery and LV remodeling after revascularization in patients with ischemic heart disease, which might be helpful in clinical decision-making and risk stratification.

[Association between myocardial ADAMTS-1 expression and myocardial fibrosis in a murine model of viral myocarditis].

OBJECTIVE: To investigate the association between myocardial ADAMTS-1 expression and myocardial fibrosis in coxsackievirus B(3) (CVB(3))-induced acute and chronic murine myocarditis model. METHODS: Balb/c mice were infected with CVB(3) (single injection or monthly injection for 3 months) to establish acute or chronic myocarditis model. Normal controls received equal-volume Eagles minimal essential medium (EMEM) without CVB(3). Hearts were examined at 7 days or 3 months post CVB(3) infection. Heart slides were stained with collagen specific picrosirius red staining and the collagen volume fraction (CVF) was calculated with image analysis software. The expressions of ADAMTS-1 were determined by RT-PCR and immunohistochemistry. RESULTS: Compared with controls, the CVF levels and myocardial expressions of ADAMTS-1 were significant increased in two myocarditis groups, especially in mice with chronic myocarditis (P < 0.01). The increased expression of ADAMTS-1 was located in endochylema as visualized by immunohistochemistry. Myocardial ADAMTS-1 mRNA was positively correlated with CVF in both myocarditis groups (r(7 days) = 0.65, P < 0.05; r(3 months) = 0.73, P < 0.01). CONCLUSIONS: ADAMTS-1 increased in proportion with collagen accumulation in acute and chronic myocarditis, which might play an important role in the development of myocardial fibrosis by modulating the collagen metabolism.

Advanced glycosylation end products might promote atherosclerosis through inducing the immune maturation of dendritic cells.

OBJECTIVE: Both advanced glycosylation end products (AGEs) and dendritic cells (DCs) have been shown to play a causative role in atherosclerosis. However, whether they function interactively in the process remains uncertain. We therefore studied the effects of AGE-bovine serum albumin (AGE-BSA) on the maturation of DCs and the expressions of scavenger receptor-A (SR-A) and receptor for AGEs (RAGE) on DCs. METHODS AND RESULTS: AGE-BSA induced DCs maturation accompanied with increased expressions of CD1a, CD40, CD80, CD83, CD86, and MHC class II. The capacity of DCs to stimulate T-cell proliferation and secretion of cytokines (interferon [IFN], IFN-gamma, interleukin [IL]-10 and IL-12) was also enhanced by AGE-BSA. AGE-BSA significantly upregulated SR-A and RAGE expression on DCs and the upregulation was abolished by inhibition of mitogen-activated protein (MAP) kinase Jnk, but not by that of Erk and p38 MAP kinase. AGE-BSA-induced expression of CD83 and secretion of IL-12 were partly inhibited by either an anti-RAGE neutralizing antibody or a Jnk inhibitor. CONCLUSIONS: AGE-BSA induces maturation of DCs and augmented their capacity to stimulate T-cell proliferation and cytokine secretions possibly through upregulation of RAGE and SR-A, which at least in part through Jnk. These findings might explain in part the interactive roles of AGEs and DCs in the processes of atherosclerosis.

Elevated matrix metalloproteinase expression after stent implantation is associated with restenosis.

OBJECTIVE: Matrix metalloproteinases (MMPs) play a key role in intimal growth and is responsible for ventricular remodeling after stent implantation. However, little is known about the relationship between early MMPs expression post-stent implantation and follow-up restenosis. METHODS: We investigated the serial changes of serum MMP-9, MMP-2 and tissue inhibitor of metalloproteinase-1 (TIMP-1) in 16 control subjects with normal coronary angiography (control) and 40 patients before and on the 1st, 3rd and 7th day after uncomplicated stent implantation. Follow-up angiography was performed at 6 months after stent implantation. RESULTS: Serum MMP-2 level was higher in patients with restenosis on the 1st day post-stent implantation and returned to pre-operation level thereafter. Serum MMP-9 levels consistently increased in patients with restenosis up to 7th day post-stent implantation; MMP-9 levels in the 1st, 3rd and 7th day after stent implantation were positively correlated to the late loss index 6 months after stent implantation. CONCLUSIONS: Increased serum MMP-9 level is associated with increased risk of restenosis post-stent implantation.

Assessment of left ventricular systolic synchronicity by real-time three-dimensional echocardiography in patients with dilated cardiomyopathy.

BACKGROUND: Recent advances in real-time three-dimensional echocardiography (RT3DE) offer the potential to assess the left ventricular (LV) dyssynchrony simultaneously by analyzing the 17 segments time-volume curves. The purpose of this study was to test the feasibility and accuracy of RT3DE for quantitative evaluation of left ventricular systolic synchronicity. METHODS: Twenty-four patients with dilated cardiomyopathy (DCM) and twenty-five healthy volunteers were enrolled in this study. Full volume RT3DE was performed by using Philips IE33 with X3-1 probe. The global and 17-segmental time-volume curves were obtained by the on-line Qlab software (version 4.2). The time to minimal systolic volume in each segment (T(msv)) was taken to derive the following indexes of systolic asynchrony: T(msv) 16-SD, T(msv) 16-Dif, T(msv) 12-SD, T(msv) 12-Dif, T(msv) 6-SD and T(msv) 6-Dif, which meant the standard deviation or the maximal difference of T(msv) among the 16, 12 and 6 segments of the left ventricle respectively. The software also provided with each of the above parameters as a percentage of the cardiac cycle. RESULTS: T(msv) 16-SD, T(msv) 12-SD and T(msv) 6-SD were all significantly larger in the DCM group than those of the control group [T(msv) 16-SD: (52.9 +/- 40.6) ms vs (8.8 +/- 6.2) ms; T(msv) 12-SD: (29.5 +/- 30.8) ms vs (6.9 +/- 4.0) ms; T(msv) 6-SD: (28.9 +/- 34.6) ms vs (7.0 +/- 4.7) ms, all P < or = 0.001]. T(msv) 16-Dif, T(msv) 12-Dif and T(msv) 6-Dif were also significantly larger in the DCM group. There were close negative relations between the LVEF determined by RT3DE and each of the indexes of systolic asynchrony, among which the indexes of T(msv)-16-SD% and T(msv)-16-Dif% correlated most closely (r = -0.703 and r = -0.701, respectively). The DCM patients had significantly larger EDV and ESV, with significantly reduced LVEF compared with the healthy subjects. CONCLUSION: RT3DE provides a simple, useful and unique approach to assess the systolic synchronicity of all the left ventricular segments simultaneously.

[Role of novel risk factors in predicting risk of ischemic cardiovascular diseases in middle aged men in twenty years in Shanghai].

OBJECTIVE: To examine the existing Framingham Risk Score (FRS) and Chinese Risk Score (CRS) in predicting the development of ischemic cardiovascular diseases (ICVD), and determine potential added value of novel risk factors. METHODS: The China Multi-Provincial Cohort Study (CMCS) was a population-based prospective cohort study in 11 provinces of China. An annual follow up was conducted in 840 men aged 35 to 64 years in Shanghai cohort, who were without coronary heart disease and stroke at baseline examination in 1992, to collect the incidence data of ICVD events (coronary death, myocardial infarction, and ischemic stroke). The detection of novel risk factors were conducted for the cohort in 2007. The basic Framingham and Chinese prediction scores power were assessed by using C-statistic of ICVD events associated with risk scores, then the novel risk factors were evaluated by adding them independently to the basic Chinese models. The area under the curve (AUC), net reclassification improvement (NRI), and integrated discrimination improvement(IDI) were calculated to determine if each of the novel risk factors improved risk prediction. RESULTS: By the end of December 2014, 24 cases of coronary heart disease (myocardial infarction or/and coronary death), 45 cases of ischemic stroke had occurred in 840 subjects in Shanghai cohort with a follow-up of 22.3 years averagely. Both the FRS and CRS had predicting power for ICVD, the AUCs were 0.6576 (95%CI: 0.5942-0.7240) and 0.7265 (95%CI: 0.6643-0.7887), respectively. The incremental AUC was 0.0689 (95%CI: 0.0196-0.1171) (P=0.006). None of the novel risk factors significantly improved the AUC. High-sensitive-CRP (hs-CRP) was the only novel risk factor resulting in a significant increase of NRI. CRS in 2007 significantly improved the IDI, but net changes were small. CONCLUSIONS: CRS had high power in the 20-year risk prediction for ICVD in middle-aged men in Shanghai. The inclusion of hs-CRP could make some improvement in risk prediction, but is unlikely to be meaningful when reclassification or new discrimination strategy are made which can change the clinical risk.

Human serum preß1-high density lipoprotein levels are independently and negatively associated with coronary artery diseases.

BACKGROUND: Serum preß1-high density lipoprotein (preß1-HDL) was defined by two-dimensional non-denaturing linear gel electrophoresis and apolipoprotein A-I immuno-blotting. Serum preß1-HDL seems to play an important role in reverse cholesterol transport, a well-known anti-atherosclerosis process. However, there are still debatable questions for its quantification and coronary artery disease (CAD) relevance. METHODS: We isolated the preß1-HDL using a new native polyacrylamide gel electrophoresis (PAGE) system and lipid pre-staining serum. We established a two-demensional gel electrophoresis system. RESULTS: We measured the preß1-HDL in Tangier disease patients and subjects with cholesterol ester transfer protein (CETP) mutation. The preß1-HDL is clearly separated from lipid-free apoA-I monomer and cannot be converted into other HDL particles under lecithin-cholesterol acyltransferase (LCAT) inhibition. This preß1-HDL is a spheroidal particle with the highest apoA-1/cholesterol ratio and highest density (≥1.21 g/ml), as compared with all other HDLs. Importantly, we found that serum from subjects with Tangier disease or with cholesterol ester transfer protein (CETP) mutation have no detectible preß1-HDL particles. We recruited a total of 102 subjects underwent diagnostic coronary angiography and measured their preß1-HDL levels. Among them, 56 had no stenosis of coronary artery and 46 were diagnosed as CAD, which was predefined as the presence of a luminal diameter stenosis ≥50 % in at least 1 major coronary artery territory. We found that preß1-HDL is independently and negatively associated with the severity of the coronary artery stenosis (Gensini score). CONCLUSION: We established a novel and simple method for human serum preß1-HDL quantification. We found that human lower preß1-HDL is an independent predictor for severer coronary artery stenosis.