Ma Yitong

GW24-e0604 Serum uric acid levels are associated with high blood pressure in Chinese children and adolescents aged 10-15 years

Objectives Uric acid is associated with cardiovascular disease such as coronary artery disease and hypertension in adults. However, among the Chinese children and adolescents, whether the uric acid was associated with the high blood pressure remained unknown. Therefore, this present study examined the association between uric acid levels and high blood pressure in a multi-ethnic study of Chinese children and adolescents using data from Xinjiang Congenital Heart Disease Survey (XCHDS). Methods The young participants aged 10 to 15 years from the XCHDS were enrolled in this study. 3 logistic regression models were conducted to investigate the relationship between the high blood pressure and uric acid levels. The concentrations of uric acid between participants with or without the high blood pressure were calculated and compared. Results A total of 3778 participants were included in the present study. The percentages of the high blood pressure in the four different uric acid quartiles were 7.4%, 8.6%, 9.6% and 11.8% respectively. In model 1, 2 and 3 of the logistic regression, the participants in the third and fourth uric acid quartiles had significantly higher chance of suffering the high blood pressure when compared with the participants in the first uric acid quartile (OR = 1.601, 1.627, 1.613, P = 0.006, 0.011, 0.013 respectively between participants in the first quartile and the third quartile; OR = 1.982, 1.962, 1.829, P = 0.001, 0.002, 0.006 respectively between participants the first quartile and the fourth quartile). The concentrations of serum uric acid were 220.67 ± 72.00 μmol/L in participants with the high blood pressure and 204.07 ± 64.32 μmol/L in participants without the high blood pressure (P = 0.024). Conclusions Among Chinese children and adolescents, increasing levels of serum uric acid are associated with high blood pressure.

GW24-e0606 Dyslipidemia in Xinjiang Uygur Autonomous Region, China

Objectives The aim of this study was to estimate the status of dyslipidemia awareness, treatment, and control in Xinjiang, China. Methods A multiple-ethnic, community-based, cross-sectional study of cardiovascular risk factors (the Cardiovascular Risk Survey [CRS]) was conducted in Xinjiang (northwestern China) between October 2007 and March 2010. We used a stratified sampling method to select a representative sample of the general population, including Chinese Hans, Uygurs, and Kazaks, in this geographic area. Seven cities (Urumqi, Kelamayi, Hetian, Zhaosu, Fukang, Tulufan, and Fuhai) were chosen and, based on the government records of registered residences, one participant was randomly selected from each household. The eligibility criterion for the study was > 35 years of age. Results A total of 14,618 participants (5,757 Hans, 4,767 Uygurs, and 4,094 Kazakhs), were randomly selected from 26 villages in 7 cities and were invited to participate. Participants with incomplete data were excluded; thus, 13, 609 participants (5,326 Hans, 4,448 Uygurs, and 3,835 Kazaks), 35-74 years of age, were analysed. We found that 7,187 participants had dyslipidemia. Among the participants with dyslipidemia, only 45.90% were aware of their serum lipid levels, 21.98% were taking lipid-lowering medication, and 17.00% achieved target serum lipid concentrations. In Han, 53.84% were aware of their serum lipid levels, 19.42% were taking lipid-lowering medication, and 16.92% achieved target serum lipid concentrations. In Uygur, 42.05% were aware of their serum lipid levels, 27.54% were taking lipid-lowering medications, and 16.23% achieved target serum lipid concentrations. In Kazak, 37.24% were aware of their serum lipid levels, 19.91% were taking lipid-lowering medications, and 18.01% achieved target serum lipid concentrations. Conclusions Dyslipidemia is highly prevalent in Xinjiang. The proportion of participants with dyslipidemia who were aware, treated, and controlled is unacceptably low. These results underscore the urgent need to develop national strategies to improve the prevention, detection, and treatment of dyslipidemia in Xinjiang (northwestern China).

GW24-e0623 Secreted frizzled-related protein-1 attenuates staurosporine-induced apoptosis in cardiomyocytes

Objectives Recently there is accumulating evidence that the wnt/frizzled pathway may play a distinct role in cardiomyocytes apoptosis. We have demonstrated that staurosporine induces cardiomyocytes apoptosis in vitro. FrzA/sFRP-1, a secreted frizzled-related protein and antagonist for the wnt/frizzled pathway. This study was to explore the role of wnt/frizzled signalling pathway in staurosporine-induced apoptosis in cardiomyocytes and assessed the hypothesis that FrzA overexpression could attenuates staurosporine-induced apoptosis in cardiomyocytes. Methods We found that the staurosporine induced the expression of Dvl-1 and subsequent up-regulation of b-catenin, which are the downstream members of wnt/frizzled pathway when cardiomyocytes apoptosis occurred. The staurosporine concentration elevated, apoptosis becomes serious and Dvl-1/b-catenin expression enhanced. Then cardiomyocytes were transfected with a recombinant AAV9 vector to deliver the FrzA gene, we found that FrzA gene suppression the expression of Dvl-1 and b-catenin and the activity of the Wnt/ frizzled pathway. Results FrzA overexpression decreased the apoptotic rate, caspase-3 activity, and the Bax/Bcl-2 ratio in cardiomyocytes treated with staurosporine. Conclusions Overexpression of FrzA inhibited the activity of the Wnt/frizzled pathway and reduced the apoptosis of cardiomyocytes.

GW24-e0980 A novel polymorphism of the CYP2J2 gene is associated with coronary heart disease in Uygur population in China

Objectives Cytochrome P450 (CYP) 2J2 is expressed in the vascular endothelium and metabolises arachidonic acid to biologically active epoxyeicosatrienoic acids (EETs). The EETs are potent endogenous vasodilators and inhibitors of vascular inflammation. The aim of the present study was to assess the association between the human CYP2J2 gene polymorphism and coronary artery disease (CAD) in a Han and Uygur population of China. Methods We use two independent case-control studies: a Han population (206 CAD patients and 262 control subjects) and a Uygur population (336 CAD patients and 448 control subjects). All CAD patients and controls were genotyped for the same three single nucleotide polymorphisms (SNPs) (rs890293, rs11572223 and rs2280275) of CYP2J2 gene by a Real-time PCR instrument. Results In the Uygur population, for total, the distribution of SNP3 (rs2280275) genotypes showed a significant difference between CAD and control participants (P = 0.048). For total and men, the distribution of SNP3 (rs2280275) alleles and the dominant model (CC vs CT + TT) showed a significant difference between CAD and control participants (For allele: P = 0.014 and P = 0.035, respectively; For dominant model: P = 0.014 and P = 0.034, respectively). The significant difference in dominant model was retained after adjustment for covariates (OR: 0.279, 95% confidence interval [CI]: 0.176-0.440, P < 0.001; OR: 0.240, 95% CI: 0.128-0.457, P <0.001, respectively). Conclusions The CC genotype of rs2280275 in CYP2J2 gene could be a protective genetic marker of CAD and T allele may be a risk genetic marker of CAD in the Uygur population in China.

GW24-e0988 A novel polymorphism of the CYP4F2 gene is associated with acute coronary syndrome

Objectives CYP4F2 is responsible for metabolising arachidonic acid to 20-hydroxyeicosatetraenoic acid (20-HETE), which plays a crucial role in the regulation of cardiovascular homeostasis. The present study aims to evaluate the association between the CYP4F2 gene polymorphism andacute coronary syndrome (ACS). Methods Four CYP4F2 SNPs were genotyped (rs1558139, rs3093166, rs3093194, rs2108622) by the TaqMan® SNP Genotyping Assay in Real-Time PCR System. We examined the association between the four SNPs and ACS using a haplotype-based case-control study that included separate analysis of the two ethnic groups, one was Han population (326 ACS patients and 338 control subjects) and the other was Uygur population (265 ACS patients and 276 control subjects). Results GG + GA genotype carriers of SNP4 (rs2108622) were more frequent among ACS patients than among controls not only in the Han population (97% vs. 92%) but also in the Uygur population (94% vs. 88%). After adjustment of confounding factors such as hypertension, diabetes, smoking, body mass index, triglyceride, the odds ratio (OR) for GG + GA genotype carriers of the rs2108622 in ACS was 2.325 (95% confidence interval:1.067-5.067) in the Han population and 2.220 (95% CI: 1.060-4.649) in the Uygur population. For the Uygur population, the frequency of the A-G-G haplotype was significantly higher for ACS patients than for control subjects (p = 0.046) Conclusions The GG + GA genotype of the rs2108622 in CYP4F2 gene may be a genetic marker of ACS in the Han and Uygur population. The T-G-G haplotype might be a dangerous genetic marker for ACS in the Uygur population of China.

ASSA13-03-40 The Study of Cardiomyocytes Gene Modified by Recombinant Adeno-Associated Virus-9 Combined with PDGF-B in Vitro

Objective To explore the feasibility, safety and anti-apoptosis of using recombinant adeno-associated virus-9 which contained platelet-derived growth factor-B (rAAV9-PDGF-B) transduction to rat cardiomyocytes in vitro. Methods The primary neonatal rat cardiomyocytes were isolated and cultured, and cardiomyocytes were transduced with rAAV9-PDGF-B at MOI (multiplicities of infection) = 5 × 105, and collected cardiomyocytes from different times after transduction, respectively at1, 3, 6, 14, 21, 28 days. Proteins were extracted from the cells for Western Blot analyse and observed the expression of PDGF-B, the transfection efficiency were measured by cell immunofluorescence. Alamar Blue assays were used to evaluate the safety of rAAV9-PDGF-B transduction to cardiomyocytes. And explore the cardiomyocytes which over expressed PDGF-B whether own the capacity of protecting cells against H2O2 –induced apoptosis by the test of apoptosis-related proteins and the ratio of Tunel-positive cell. Results Cardiomyocytes were over expressed PDGF-B at the third day after transduction, and the expression of PDGF-B was increased gradually. After days 5, the expression would be stable and sustaining, such expression can last for 28 days. Cell immunofluorescence showed that approximately 78.6% cardiomyocytes overexpressed PDGF-B at days 6 after transduction. Alamar Blue assays, which evaluated the toxicity of rAAV9-PDGF-B transduction to cardiomyocytes, showed that the reduction ratios of cardiomyocytes at different times were all close to 1.0. Moreover, in Cardiomyocytes which over expressed PDGF-B, H2O2 –induced apoptosis was accompanied by the down-regulation of Bax and Tunel-positive cells, by the up-regulation of Bcl-2 and P-Akt, there was no difference between group PDGF-B+ H2O2 (P > 0.05). Conclusions rAAV9-PDGF-B could effectively transduce cardiomyocytes cultured in vitro and persistently express PDGF-B at least 28 days. rAAV9-PDGF-B was no significant toxic effects on cardiomyocytes. Furthermore, the cardiomyocytes which over expressed PDGF-B also own the capacity of protecting cells against H2O2 –induced apoptosis, these effects might be mediated by phosphorylation of Akt signalling in cardiomyocytes.

ASSA13-03-50 Transfection of Rats H9C2 Cells with Recombinant Adeno-Associated Virus Serotype 9 Mediated Anti-NF-kB Ribozyme in Vitro and Effect of Nuclear Factor-kB Activity

Objective To evaluate the transfection efficiency using recombinant adeno-associated virus Serotype 9 mediated Anti-NF-κB ribozyme and enhanced green fluorescent protein (rAAV9-EGFP-R65) to rats H9C2 cells and the effect of Nuclear Factor-κB (NF-κB) activity. Methods rAAV9-EGFP-R65 was transfected into H9C2 cells at multiplicities of infection (MOI = 1 × 106 v.g./cell). EGFP expression in the cells was observed under inverted fluorescence microscope, and the EGFP-positive cell percentage was determined by flow cytometry. Alamar Blue assay was used to assess the proliferation of the transfected cells. H9C2 cells were treated with TNF-α, rAAV9-EGFP- R65 and PDTC. The DNA binding activity of NF-κB was examined by electrophoretic mobility shift assay (EMSA). Results The cells with rAAV9-EGFP-R65 transfection at MOI of 1 × 106 v.g./cell began to exhibit EGFP expression 1 day after transfection. The fluorescence intensity increased with the time of transfection. EGFP expression reached the maximum on day 5, at the point of which the transduction efficiency of rAAV9-EGFP-R65 inH9C2 cells was (32.27 ± 3.19)%. Alamar Blue assay did not reveal significant difference in the absorbance between the transfected cells and the control cells. TNF-a could active NF-κB, rAAV9- EGFP-R65 and PDCT can efficiently decrease NF-κB activation in rats H9C2 cells. Conclusions rAAV9-EGFP-R65 can be stably and efficiently expressed in H9C2 cells without causing cell growth inhibition. rAAV9-EGFP-R65 can availably inhibit NF-κB activation in rats H9C2 cells in vitro. This study played foundation for further research.

WHAT IMPACTS DOOR-TO-BALLOON TIME–AN ANALYSIS FROM A SINGLE CENTRE IN A TERTIARY CARE GENERAL HOSPITAL

Objectives Current data have shown that many factors impact Door-to-ballon (DTB) time in the patient of ST-elevation myocardial infarction (STEMI). However, major factors are diverse in different region of china. Our study was aim to analyse the impact factors which significantly prolonged the DTB time in our hospital. Methods We analysed the DTB time and its components from January 2008 to December 2010 in 301 consecutive patients presenting with STEMI. Then, we determined which factors significantly prolonged the DTB time. Results The median DTB time of all the patients was 149±78 min, the group was divided by DTB time, ≤120 min group and>120 min group. The median DTB time of two groups were 87±29 min and 201±68 min respectively. The components of DTB time included that the time of diagnosis in ED (21±7 vs 22±4, p>0.05), the time of consultation of cardiologist (19±8 vs 50±21, p=0.000), the time of explaining condition (16±7 vs 86±42, p=0.000), transferred to catheterisation laboratory (12±5 vs 13±3, p>0.05), preparation of catheterisation laboratory in working hours (8±2 vs 9±2, p>0.05), preparation of catheterisation laboratory in on-call hours (40±6 vs 42±8, p>0.05). Besides, there were more STEMI patients presenting to hospital during working hour in ≤120 min group (31.9% vs 63.2%, p<0.05). Conclusions In our tertiary care general hospital, the time of consultation of cardiologist and explaining condition really account for the prolonged DTB time. What is more, the patients presented to hospital during working hour may shorten the DTB time. Therefore, directly awaking the catheterisation laboratory by emergency department and promoting the cognitive level of primary PCI in public may shorten DTB time.

Relationship between a novel polymorphism of the C5L2 gene and coronary artery disease

Background C5L2 has been demonstrated to be a functional receptor of acylation-stimulating protein (ASP), which is a stimulator of triglyceride synthesis or glucose transport. However, little is known about the variations in the coding region of the C5L2 gene and their association with coronary artery disease (CAD). Methodology/Principal findings The authors identified a novel single nucleotide polymorphism (SNP), 698C>T (P233L), in exon 2 using a PCR direct-sequencing method. This nucleotide change causes the amino-acid order from proline to leucine at codon 233. We examined the role of this SNP for CAD using two independent case–control studies: one was in the Han population (492 CAD patients and 577 control subjects) and the other was in the Uygur population (319 CAD patients and 554 control subjects). Heterozygote carriers of the 698CT genotype were more frequent among CAD patients than among controls in the Han population (7.3% vs 1.7%) and in the Uygur population (4.7% vs 1.6%). The odds ratio (OR) for carriers of the 698CT genotype for CAD was 4.484 (95% CI: 2.197 to 9.174) in the Han group and 2.989 (95% CI: 1.292 to 6.909) in the Uygur population. After adjustment of confounding factors such as sex, age, smoking, alcohol consumption, hypertension, diabetes, as well as serum levels of triglyceride, total cholesterol, high-density lipoprotein, the difference remained significant in the Han group (p<0.001, OR=6.604, 95% CI: 2.776 to 15.711) and in the Uygur group (p=0.047, OR=2.602, 95% CI: 1.015 to 6.671). Conclusion/Significance The 698CT genotype of C5L2 may be a genetic marker of CAD in the Han and Uygur population in western China.

e0054 Recombinant adeno-associated virus serotype 9 transfection of rats H9C2 cells in vitro

Objective To evaluate del transfection efficiency using recombinant adeno-associated virus Serotype 9 mediated enhanced green fluorescent protein (rAAV9- EGFP) to rats H9C2 cells and the impact on growth of H9C2 cells. Methods rAAV9-EGFP was transfected into H9C2 cells at different multiplicities of infection (MOI=1×105, 1×106, 1×107). EGFP expression in the cells was observed under inverted fluorescence microscope and the EGFP-positive cell percentage determined by flow cytometry. Alamar Blue assay was used to assess the proliferation of the transfected cells. Results The cells with rAAV9-EGFP transfection at MOI of 1×106 and 1×107 began to exhibit EGFP expression 1 days del after transfection and the cells transfection at MOI of 1×105 began to exhibit EGFP expression 2 days after transfection. The fluorescence intensity increased with the MOI used for transfection. EGFP expression reached the maximum on day 4, at the point of which the transduction efficiency of rAAV9-EGFP in H9C2 cells was (14.1±0.2)%, (35.1±4.8)% and (56.8±0.1)%. Corresponding to MOIs of 1×105, 1×106 and 1×107, respectively. Alamar Blue assay did not reveal significant difference in the absorbance between the transfected cells and the control cells after transfection. Conclusions rAAV9-EGFP gene can be del transfected in a stable manner and efficiently expressed in H9C2 cells without causing cell growth inhibition. This del The results of this study played foundation (del) (provides a platform) for further research.