Correlation analysis between visualization of hemorrheologic changes and platelet function in patients with acute coronary syndrome after percutaneous coronary intervention

AIM:To intuitionally observe the characteristics of blood rheology in the patients with acute coronary syndrome (ACS) after percutaneous coronary intervention (PCI) for 1 year to 3 years by micro-channel array flow analyzer (MC-FAN) combined with other platelet function indexes, and to explore the correlations between the test results of MC-FAN and platelet function. METHODS:This study brought 74 patients with ACS after PCI for 1 year to 3 years into test group, and 21 healthy subjects were enrolled as normal group. The levels of platelet aggregation test (PAgT), platelet adhesiveness test (PAdT), P-selectin, platelet-derived growth factor BB (PDGF-BB) and von Willebrand factor (vWF) were detected. MC-FAN HR300 was used to detect the transiting time (MC-FAN TT) of the blood passing through the model body capillaries. The differences of the test results between the 2 groups were compared, and the correlations between the results of MC-FAN and platelet function in the patients with ACS after PCI were also explored. RESULTS:Compared with normal group, the MC-FAN TT in test group was prolonged (P<0.01), the ability of erythrocyte deformation was weakened, and the leukocyte attaching the vascular wall and platelet adhesion and aggregation relatively increased. The levels of PAgT, PAdT, P-selectin and PDGF-BB in test group were all higher than those in normal group (P<0.01). No difference of vWF between the 2 groups was observed. The intergroup correlation analysis showed that there were correlations between MC-FAN TT and platelet function, in which 10 μL MC-FAN TT and 30 μL MC-FAN TT had the most significant correlation with P-selectin (r=0601, P<0.01; r=0334, P<0.01), 60 μL MC-FAN TT had the most significant correlation with PAgT (r=0527, P<0.01), and 100 μL MC-FAN TT had the most significant correlation with PAdT (r=0. 815, P<0.05). CONCLUSION:The visualization of hemorrheologic changes and platelet function in the patients with ACS after PCI are abnormal.There are correlations between MC-FAN TT and platelet function.The results of MC-FAN can objectively evaluate the blood rheology of the patients, and provide the reference for clinical treatment.     

Microparticles and acute coronary syndrome

Coronary heart disease (CHD) is an important cause of death in the world. Among them, acute coronary syndrome (ACS) is a type of CHD that has a high mortality rate. Microparticles (MPs) are tiny particles formed in the process of cell activation and apoptosis, carrying specific markers of their mother cells. There is more and more evidence that MPs derived from endothelial cells, platelets, white blood cells and other cells are closely related to the formation of ACS. Through a variety of pathways, they take part in endothelial cell damage, vascular dysfunction, plaque erosion and rupture, and thrombosis. The findings of MPs enrich our knowledge about ACS, and may provide a new approach to the treatment of ACS.     

Alteration of serum IL-6, IL-8, TNF-α and sICAM-1 in patients with pre-menstruation recurrent aphthous ulceration

AIM:To investigate the serum levels of interleeukin-6(IL-6), interleukin-8(IL-8), tumor necrosis factor-alpha(TNF-α) and soluble intercellular adhesion molecule-1 (sICAM-1)in female patients with pre-menstruation recurrent aphthous ulceration(RAU).METHODS:Serum levels of IL-6, IL-8, TNF-α and sICAM-1 in 21 pre-menstruation RAU patients were examined using ELISA technique, and compared to 10 healthy individuals and 22 the female RAU patients unrelated to menstrual cycle.RESULTS:The serum levels of IL-6, IL-8, TNF-α in patients with pre-menstruation RAU were not only significantly higher than that in the normal control group(P＜0.01), but also higher than that in the RAU patients without pre-menstruous recurrence (P＜0.01).The level of serum TNF-α in the RAU patients without pre-menstruous recurrence was slightly higher than that in the normal control group(P＜0.05).The level of sICAM-1 had not changed.CONCLUSION:The serum levels of IL-6, IL-8 and TNF-α increase in pre-menstruation RAU patients, which might play a role in local lesion of RAU.     

Effects of ambroxol combined with low-dose heparin on production of ICAM-1 and E,P-selectin in acute lung injury

AIM: To study the production of intercellular adhesion molecule-1(ICAM-1), E-selectin and P-selectin in serum, lung tissues and bronchoalveolar lavage fluid(BALF)of acute lung injury(ALI) model and to observe the effects of ambroxol combined with low-dose heparin on the changes of the 3 factors above.METHODS: Twenty-four healthy rabbits were randomly divided into 3 groups: normal saline control group (NC), oleic acid injury group (OA), ambroxol+ heparin treatment group (AH). The rabbit ALI model was induced by oleic acid injection through auricular vein. Partial pressure of O₂ in artery(PaO₂) was analyzed.The concentrations of ICAM-1 and E-selectin were detected by ELISA.The apoptosis index(AI) was measured by TUNEL method.The expression of P-selectin was determined by immunohistochemical method.The ultrastructural changes of the lung tissues were observed under electron microscope, and the lung wet/dry ratio(W/D) was calculated.RESULTS: PaO₂ in AH group and OA group was significantly lower (P<0.01) than that in NC group, and PaO₂ in AH group was significantly higher than that in OA group (P<0.01). The concentrations of ICAM-1 and E-selectin in serum, lung tissues and BALF, and AI and W/D in lung tissues in AH group were higher (P<0.05 or P<0.01) than those in NC group, and was lower than those in OA group (P<0.05 or P<0.01). In NC group, no significant change of the above parameters at all time points was observed (P>0.05). In OA group, PaO₂ was significantly decreased (P<0.01) with the pathological process developed, and the concentrations of ICAM-1 and E-selectin were significantly increased. In AH group, PaO₂ was decreased (P<0.05),and the concentrations of ICAM-1 and E-selectin were increased with the process of ALI developed. The P-selectin expression in lung tissues of OA group was distributed mainly in inflammatory cells, capillary endothelial cells and plasma. From low to high levels, the order was NC group < AH group < OA group in the expression of P-selectin. The most obvious apoptosis was observed in OA group. No apoptosis or occasional positive cells were found in NC group. The apoptotic rate in AH group was significantly reduced compared with that in OA group.CONCLUSION: In ALI induced by OA, ICAM-1, E-selectin and P-selectin are significantly increased and are involved in the occurrence and development of ALI. Ambroxol combined with low-dose heparin reduces the levels of ICAM-1, E-selectin and P-selectin, the pulmonary edema and the lung injury, improves pulmonary functions, and plays an important role in the prevention and treatment of acute lung injury.     

Effect of lipid peroxidation on expression of vascular cell adhesion molecule-1 in cultured human endothelial cells

AIM:To observe the expression of vascular cell adhesion molecule-1 (VCAM-1) in cultured human umbilical vein endothelial cells (HUVEC) by lipid peroxidation injury induced by exposure to diamide.METHODS:Expression of VCAM-1 mRNA and protein in HUVEC was determined by in situ hybridization and a cell enzyme-linked immunosorbent essay (cell ELISA), respectively.RESULTS:The HPIAS-1000 image analytic system in situ hybridization detected that the mean absorbance values in experiment groups(1, 5 and 10 μmol/L diamide for 8 hours) were 0.147±0.013, 0.292±0.020 and 0.396±0.022, which were 1.91-fold, 3.79-fold and 5.14-fold as much as that of the control group (0.077±0.011), respectively. There was significant statistical difference between groups (P＜0.01). The cell ELISA showed that the mean absorbance values of VCAM-1 proteins in experiment groups were 0.158±0.008, 0.220±0.017 and 0.321±0.023, which were 1.53-fold, 2.12-fold and 3.09-fold as much as that of the control group (0.104±0.016), respectively. The analysis of variance proved the significant difference between groups (P<0.01).CONCLUSION:The results suggest that the increased expression of VCAM-1 is integral in promoting adhesion of monocytes to the vascular endothelium.     

Inhibitory effects of Angelicae sinensis and sodium ferulate on acute inflammatory liver injury and expression of ICAM-1 and E-selectin in mice

AIM: To explore the effects of Angelicae sinensis preparation and sodium ferulate on inflammatory liver injury induced by lipopolysaccharide (LPS) and their molecule mechanism. METHODS: ICR mice were divided into five groups: Angelicae sinensis group, sodium ferulate group, dexamethasone group, inflammation control group and normal group. The model of inflammatory injury in mice was set up by tail vein injection with the bacillus calmette-guerin (BCG) and LPS respectively prior and posterior to administration of those tested drugs. The tested drugs (the preparations of Angelica sinensis, sodium ferulate and dexamethasone) and normal saline were given respectively to the corresponding group. The pathological observation of the liver tissues in mice was made for the intensity of inflammatory liver injuries. The immunohistochemical detection and comparison were performed for the expression of ICAM-1 and E-selectin proteins in the liver tissues in those mice. RESULTS: The intensities of liver inflammatory injuries in mice from drug-treated groups were obviously lighter than that from the inflammation control group (P<0.01). The expressions of ICAM-1 and E-selectin proteins in the liver tissues of mice from the inflammation control group were not only significantly higher than that in normal control, but also obviously higher than that from drug-treated groups (P<0.01). CONCLUSIONS: Angelica sinensis and sodium ferulate alleviate inflammatory liver injury induced by injection of LPS. Their suppressive effects on inflammatory liver injury may be associated with the decrease in the expression of ICAM-1 and E-selectin proteins.     

Complement activation in acute coronary syndromes

AIM: To evaluate complement activation in patients with all forms of acute coronary syndromes (ACS) and to examine the relationship between the degree of complement activation and myocardial injury.METHODS: The subjects were divided into 2 groups: 110 ACS patients (group ACS) and 18 healthy persons (group control).One hundred and ten patients with ACS were divided into 3 sub-group: 51 patients with ST-segment elevated myocardial infarction (STEMI),28 patients with non-ST-segment elevated myocardial infarction (NSTEMI) and 31 patients with unstable angina (UA).Complement 3 (C₃),complement 4 (C₄),troponin T (TnT) as well as creatine kinase MB (CK-MB) were evaluated.RESULTS: Plasma C₃ and C₄ peak levels were significantly higher in patients with STEMI ［(1 525±302)mg/L and (423±123) mg/L］ and NSTEMI ［(1 516±289)mg/L and (396±68) mg/L］ than those in patients with UA ［(1 275±172)mg/L and (356±91) mg/L］ and the control subjects ［(1 072±196)mg/L and (182±73) mg/L］ (P<0.01 for all).Also,C₃ and C₄ serum levels in patients with UA were significantly higher than those in control subjects (P<0.01 for all).At one-week follow-up,plasma levels of C₃ and C₄ were significantly different among various days in patients with ACS (P<0.01).Plasma C₃ and C₄ levels in ACS showed a relationship with peak creatine kinase MB (CK-MB) and troponin T (TnT) levels (P<0.01).CONCLUSION: Plasma C₃ and C₄ levels are elevated in ACS in present study.The relationship between C₃,C₄ levels and ACS suggests that the complement activation is related to necrosis within the myocardium.     

Activation of vascular endothelial cells improves the homing of hematopoietic stem cells

AIM: To study the effect of endothelial cell activation on the homing of hematopoietic stem cells (HUHSC) during transplantation. METHODS: Human umbilical vein endothelial cells (HUVEC) were cultured to single layer and activated by vascular endothelial growth factor (EVGF), granulocyte colony stimulating factor (G-CSF) and lipopolysaccharide (LPS), respectively. The HUHSC, enriched by eliminating red blood cells, granulocytes, monocytes and lymphocytes from cord blood, were cocultured with activated HUVEC to make adhesion. The adhesive ability of activated HUVEC to C-Kit⁺ HUHSC was assayed by ELISA. Anti-VCAM-1 monoclonal antibody was used to detect the effect of activation on HUVEC and HUHSC interactions. RESULTS: Resting HUVEC had a little adhesive ability to HUHSC. A great enhancement of adhesive ability was showed when HUVEC was activated by VEGF, G-CSF and LPS. In the presence of anti-VCAM-1, the adhesive ability of activated HUVEC was decreased remarkablely. CONCLUSION: HUHSC homing may be related to the activation of endothelial cells and adhesion molecules.     

Serum acute phase reactants sCRP,SAA, IL-6 in acute coronary syndrome and its clinical significance

AIM: To investigate the possible involvement of CRP, SAA, IL-6 and its significance in acute coronary syndrome. METHODS: Serum SAA, IL-6 and sCRP levels in 40 patients with AMI, 40 patients with UAP and 40 controls were measured by means of ELISA. RESULTS: Serum SAA, IL-6 and CRP were significantly higher in ACS group (AMI or UAP) than in control group. In the Spearman correlation analysis , we found a correlation among IL－6, SAA, sCRP, fibrinogen both in the AMI group and UAP group. There was no correlation between CK-MB and serum SAA, IL-6 and sCRP levels either in AMI group or in UAP group. In the AMI group, there was no difference of serum SAA,IL-6 and sCRP levels when divided into ≤6 score group and >6 score group using Wagner's QRS scoring system. CONCLUSIONS: Serum levels of certain inflammatory markers have some diagnostic value for ACS. The high levels of serum SAA, IL-6 and sCRP are not mainly caused by myocardial death, but by the inflammation in the multifocal unstable plague.     

Prognostic value of serum concentration of heart-type fatty acid-binding protein on acute coronary syndrome at early stage

AIM: To investigate the prognostic value of serum concentration of heart-type fatty acid-binding protein (H-FABP) at early stage of acute coronary syndrome (ACS). METHODS: This was a prospective observational study with a follow-up of 12 months. The H-FABP was measured within 6 h after onset of chest pain in 172 consecutive patients hospitalized for ACS. The study endpoints were cardiac events, which were defined as cardiac death, subsequent nonfatal acute myocardial infarction (AMI), nonfatal heart failure and recurrent angina. RESULTS: During the period of 12-month follow-up, 47 cardiac events, including 6 cardiac deaths, 9 nonfatal AMIs, 11 nonfatal heart failures and 21 recurrent anginas, were observed. The concentrations of H-FABP in the patients with cardiac events were significantly higher than those in the patients without cardiac events (P<0.01). Meanwhile, the cardiac events were more prevalent in increased H-FABP (above the median of 12.5 μg/L) group (36 events vs 11 events, P<0.01). In a Cox proportional hazards model that adjusted for baseline variables including demographics, clinical characteristics, ST deviation and troponin I, increased H-FABP (above the median of 12.5 μg/L) was independently associated with cardiac events in all patients. CONCLUSION: Elevation of H-FABP is associated with an increased risk of cardiac events in patients presenting across the spectrum of ACS and is independent of other established clinical risk predictors and biomarkers.     

Colquhounia root tablet inhibits the expression of adhesion molecule in acute lung injury of rats

AIM: To study the effects of colquhounia root tablet on the expression of adhesion molecule in acute lung injury of rats.METHODS: The rats were divided into 3 groups: ALI group,colquhounia root tablet＋ALI group and control group .ALI animal model was performed by treatment with oleic acid.The positive expression rates of CD11a,CD11b and CD18 in polymorphonuclear neutrophils and monocytes were analyzed by flow cytometry.ICAM-1 expression in lung tissue was determined by immunohistochemistry,histopathological examination and biological markers were measured from lung specimens.RESULTS: Colquhounia root tablet decreased the expression of CD11a,CD11b and CD18 in polymorphonuclear neutrophils and monocytes,and ICAM-1 in lung tissue (P<0.01),pathologic changes and the biological markers of ALI significantly ameliorated.CONCLUSION: The increase in the expression of CD11a/CD18 and CD11b/CD18 in polymorphonuclear neutrophils and monocytes,and expression of ICAM-1 in lung tissues of ALI may be involved in the formation of ALI.Colquhounia root tablet effectively ameliorates the lung damage,mechanism of which may be related to inhibition of adhesion molecule expression.     

Influences of bypass-activated complement and TNF-α on endothelial cells to express ICAM-1 and PMN-EC adhesion

AIM:To investigate the influences of bypass-activated complement and TNF-α on intercellular adhesion molecule 1(ICAM-1) expression in endothelial cells, and polymorphonuclear neutrophil-endothelial cell (PMN-EC) adhesion.METHODS:In vitro, zymosan-activated human serum(ZAHS)and/or TNF-Adirectly stim-ulated the HUVECS monolayers.PMN-EC adhesion percentage was measured, and modified whole-cell ELISA was used for measurement of ICAM-1.RESULTS:ZAHS alone resulted in no significant change in the degree of PMN adhesion and the level of ICAM-1. Activation of HUVECS with TNF-α followed by ZAHS stimulation resulted in greater increase in PMN-EC adhesion and ICAM-1 expression, as compared with the activation with TNF-α alone.CONCLUSION:Bypass-activated complement can promote ICAM-1 expression and neutrophil -endothelium adhesion induced by TNF-α, and so it is effective for promoting inflammation.     

Role of macrophage and intercellular adhesion molecule-1 in the pathogenesis of oleic-acid-induced rat acute lung injury

AIM: To investigate the role of infiltration of macrophages and expression of intracellular adhesion molecule-1 in the pathogenesis of oleic-acid-induced acute lung injury rats. METHODS:The rats were subjected to injection of oleic acid (oleic acid group) or saline solution (control). After injecting oleic acid or saline for 4 hours, the PaO₂ of the left heart, lung permeability index(LPI), the number of macrophage and the levels of soluble intercellular molecule-1 (sICAM-1) in the bronchial alveolar lavage fluid (BALF) were measured. The levels of expression of ICAM-1 mRNA were evaluated by in situ hybridization and the degree of macrophage infiltration and the expression of ICAM-1 were evaluated by double staining immunocytochemistry. RESULTS:The PaO₂ of the oleic acid group was significantly lower than that of the control group (P＜0.01) and the LPI of the oleic acid group was significantly higher than that of the control group (P＜0.01). The cell number of macrophage and sICAM-1 level were significantly higher in the oleic acid group than those of the control group (P＜0.01). There were marked upregulation of ICAM-1 mRNA expression in injuried lung tissue compared with the normal lung tissue. Furthermore, the infiltrated number of macrophage and the level of ICAM-1 expression showed strong positive correlation with the lung injury parameters, PaO₂ and LPI.CONCLUSION:The infiltration of macrophage may play a pivotal role in the pathogenesis of progressive lung injuries induced by intravenous oleic acid injection,ICAM-1 may mediate the infiltrat ion and adhesion of macrophage in the injuried lung t issue and contribute to the development of acute lung injury.     

Effect of vitamin E on ox-LDL-induced monocyte/endothelium adhesion

AIM:To observe the role of ox-LDL in rabbit endothelium/circulating monocyte adhesion in vitro and the effect of vitamin E.METHODS:Cultured rabbit's endothelial cells were incubated with different concentrations of ox-LDL, then incubated with rabbit's monocytes to observe cell's adhesion. With Northern blotting, endothelial VCAM-1 mRNA expression was measured. By using vitamin E incubation before ox-LDL, above steps were repeated to observe the effect of vitamin E.RESULTS:When ox-LDL concentrations were 2.5 mg/L,5 mg/L,10 mg/L,monocytes adhesive to endothelium per microscope field were 132.8±20.2,350.0±37.2,502.6±78.8,respectively They were al significantly higher than that of control group(51.2±7.7,P<0.01)Addition of vitamin Eat comcentration of 10 mol/L,20 mol/L,40 mol/L respect ively before ox-LDL incubation yield fewer adhesive monocytes as follow:422.3±32.2,296.0±21.7,205.2±36.6,respectively,but only the last two concentrations had significant effects(P<0.01)Endothelial VCAM-1 mRNA expression in ox-LDL group was higher than that in vitamin E group(0.49±0.09 vs 0,33±0.10,P<0.05).CONCLUSIONS:ox-LDL directly enhances rabbit's endothelium/monocyte adhesion by inducing expression of VCAM-1 in endothelium. Vitamin E inhibites these effects of ox-LDL.     

Expression of NGAL in serum and renal tissues of patients with primary nephrotic syndrome and acute kidney injury

AIM: To observe the changes of neutrophil gelatinase-associated lipocalin (NGAL) level in serum and renal tissues of the patients with primary nephrotic syndrome (PNS) and acute kidney injury (AKI). METHODS: Seventy-two PNS patients were selected in the study and divided into 2 groups according to the pathological results of renal biopsy.The patients in PNS+AKI group included 15 cases of PNS with acute tubular necrosis (ATN), in which there were 10 cases of minimal change disease (MCD) with ATN and 5 cases of mesangial proliferative glomerulonephritis (MsPGN) with ATN. The patients in PNS alone group included 57 cases of PNS without ATN. According to the pathological types, they were divided into MCD group (24 cases), membranous nephropathy (MN) group (23 cases) and MsPGN group (10 cases). Serum samples from 15 healthy persons and 5 cases of normal renal tissues were used as controls. The serum levels of NGAL were detected by ELISA. The distribution and expression of NGAL in the renal tissues were observed by immunohistochemical method. RESULTS: (1) The serum level of NGAL in PNS alone group (including MCD group, MN group and MsPGN group) was significantly higher than that in normal control group. Compared with MCD group and MsPGN group, the serum level of NGAL in MN group increased significantly. The renal tissue positive expression index of NGAL in MCD group, MN group and MsPGN group was significantly higher than that in normal control group. (2) The serum level of NGAL and the renal tissue positive expression index of NGAL in MCD+ATN group and MsPGN+ATN group were significantly increased compared with PNS alone group. (3) The serum level of NGAL was positively correlated with 24 h urine protein, blood urea nitrogen and serum creatinine. No relation with serum albumin and β2 microglobulin was observed. The serum level of NGAL was positively correlated with the NGAL level in the renal tissue. CONCLUSION: The serum level of NGAL can be used as one of the sensitive indexes for early and non-invasive detection of PNS with AKI.     

Neuregulin-1 induces expression of angiogenic factors in coronary artery smooth muscle cells

ATM: To investigate the effects of neuregulin-1 (NRG-1) on the expression of angiogenic factors in human coronary artery smooth muscle cells (HCASMCs). METHODS: HCASMCs were cultured in vitro, and the cells at the 3rd passage were collected to assess the expression and phosphorylation of ErbB by Western blot. After HCASMCs were cultured under normal condition, with hypoxia and serum deprivation, or with NRG-1 treatment, the expression of vascular endothelial growth factor (VEGF), angiopoietin-1 (Ang-1) and angiopoietin-2 (Ang-2) was determined by Western blot. RESULTS: The expression of ErbB2, ErbB3 and ErbB4 was observed in the HCASMCs, and the phosphorylation of these receptors was increased by NRG-1 treatment. Compared with control group, the expression of VEGF and Ang-1 in the HCASMCs was significantly increased in hypoxia and serum deprivation group (P<0.05﹚,while no difference in the expression of Ang-2 between the 2 groups was found.Compared with hypoxia and serum deprivation group,the expression of VEGF and Ang-1 in the H CASM Cs treated with NRG-1 was furtherincreased﹙P<0.05﹚,and no difference in the expression of Ang-2 between the 2 groups was observed.CONCLUSION: HCASMCs express ErbB2, ErbB3 and ErbB4, and the phosphorylation of the receptors is increased by NRG-1. Hypoxia, serum deprivation and NRG-1 treatment induce the increased expression of VEGF and Ang-1 significantly.