The optimal cutoff point of waist-to-hip ratio for screening Uyghur population aged 35 years and over at high-risk of cardiovascular diseases in Xinjiang / 中华心血管病杂志

<p><b>OBJECTIVE</b>To explore the appropriate waist-to-hip ratio (WHR) cutoffs to identify people at high risk of cardiovascular disease of Uygur population aged 35 years and over in Xinjiang.</p><p><b>METHODS</b>The cardiovascular risk survey (CRS) in Xinjiang was conducted from October 2007 to March 2010, using 4-stagestratified random sampling method and 14 618 representative participated this survey, and the questionnaire survey, anthropometric data, blood pressure, serum total cholesterol, triglyceride, low density lipoprotein (LDL-C), high density lipoprotein (HDL-C) and fasting glucose were measured. A total of 4 657 participants aged 35 years and over with complete anthropometric data were analyzed. The sensitivity, specificity and distance on the receiver operating characteristic (ROC) curve of different WHR levels predicting risk factors of cardiovascular disease were calculated. The analysis method of ROC curve was used to determine the optimum cut-off point of WHR predicting risk factors of cardiovascular disease.</p><p><b>RESULTS</b>(1) There were significantly differences in prevalence of hypertension, diabetes mellitus, hypercholesterolemia, low HDL-C level, and hypertriglyceridemia between WHR < 0.75,0.75 ≤ WHR < 0.80,0.80 ≤ WHR < 0.85,0.85 ≤ WHR < 0.90,0.90 ≤ WHR < 0.95,0.95 ≤ WHR < 1.00, WHR ≥ 1.00 in male participants (P < 0.01 or 0.05), LDL-C level was similar among groups in males (P = 0.139). There were significantly differences in prevalence of hypertension, diabetes mellitus, hypercholesterolemia and hypertriglyceridemia between WHR < 0.75,0.75 ≤ WHR < 0.80,0.80 ≤ WHR < 0.85,0.85 ≤ WHR < 0.90,0.90 ≤ WHR < 0.95,0.95 ≤ WHR < 1.00, WHR ≥ 1.00 in female participants (all P < 0.01), and there were no significantly differences in prevalence of high LDL-C level and low HDL-C level among groups in females (both P > 0.05). (2) ROC analysis for hypertension, dyslipidemia, diabetes and ≥ 2 of these risk factors suggested a WHR cutoff of 0.92 for men and 0.90 for women as the optimal cutoff value for predicting high risk of cardiovascular disease of Uygur population aged 35 years and over in Xinjiang.</p><p><b>CONCLUSION</b>Higher WHR cutoffs are needed for screening people at high risk of cardiovascular disease among Uygur population aged 35 years and over in Xinjiang.</p>

Effects of norepinephrine on proliferation and apoptosis of neonatal cardiac fibroblasts in rats / 中华心血管病杂志

<p><b>OBJECTIVE</b>To investigate the effects of different concentrations of norepinephrine (NE) on proliferation and apoptosis of cultured cardiac fibroblasts (CFBs) from neonatal mice and to elucidate related mechanisms.</p><p><b>METHODS</b>CFBs of Sprague-Dawley (SD) rats were isolated and cultured and divided into normal control group and different concentration of NE intervention groups (0.1, 1, 10, 50, and 100 µmol/L). Water soluble tetrazolium-1 (WST-1) assay was carried out to detect the viability of CFBs. Morphology of apoptosis cells was evaluated by fluorescence microscope with terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining. The expressions of collagen I, collagen III, pro-oncogene c-myc in CFBs were detected by reverse transcription-polymerase chain reaction (RT-PCR). The phospho-mitogen activated protein kinase (p-p38MAPK) and caspase3 protein levels were examined by Western blot.</p><p><b>RESULTS</b>Proliferation was significantly increased in 1 µmol/L and 10 µmol/L groups compared with the normal control group (1.05 ± 0.05 and 1.09 ± 0.02 vs. 1.00 ± 0.03, all P < 0.05).CFBs apoptosis was significantly enhanced in 50 µmol/L and 100 µmol/L groups ((22.69 ± 2.18)% and (36.40 ± 6.80)% vs.(4.50 ± 1.08)%, all P < 0.05). Expression of Collagen I peaked in 10 µmol/L group, expression of collagen III and c-myc increased dose-dependently in proportion to increasing NE concentrations (all P < 0.05 vs. control group). The expression of p-p38MAPK and caspase3 was also significantly upregulated in a dose-dependent manner in NE groups (all P < 0.05 vs. control group).</p><p><b>CONCLUSIONS</b>Low concentration NE induces CFBs proliferation and high concentration NE promotes CFBs apoptosis. p38MAPK phosphorylation may be a major mediator of NE-induced effects on CFBs.</p>

Distribution of β3-adrenergic receptors gene polymorphisms and its association with serum lipid in Han and Uighur populations in Xinjiang / 中华流行病学杂志

<p><b>OBJECTIVE</b>To investigate the distribution of β3-adrenergic receptors (ADRβ3) gene polymorphisms (rs2298423 and rs6986132) and its association with serum lipid in Han and Uighur populations in Xinjiang.</p><p><b>METHODS</b>Genotypes of the ADR β 3 gene were detected in 362 Uighur and 653 Han healthy individuals who were randomly selected in Xinjiang by real-time PCR (TaqMan) method.</p><p><b>RESULTS</b>1) Frequencies of TT, GT and GG genotypes of the rs2298423 locus were 76.5%, 22.1% and 1.4% in the Uighurs but 74.7%, 23.3% and 2.0% in the Hans. There was no significant difference noticed in distribution of genotypes between the two populations (P = 0.695). 2) Frequencies of GG, AG and AA genotypes of the rs6986132 locus were 81.5%, 16.6% and 1.9% in Uighurs but 59.6%, 33.7% and 6.7% in Hans. There was significant difference noticed in the distribution of genotypes between the two populations (P < 0.001). 3) Total cholesterol and low density lipoprotein cholesterol levels were significantly higher in GG or GT genotypes than in the TT genotype carriers of rs2298423 in the Uighur individuals (P < 0.01), but not in the Han population. Serum lipid level that including total cholesterol (TC), low density lipoproteins cholesterol (LDL-C), triglyceride and high density lipoproteins cholesterol did not show significant differences in the distribution of rs6986132 genotypes between the two populations (P > 0.05). After adjusting for factors as gender, age, height, weight, waist circumference, blood urea nitrogen, creatin, glucose, triglyceride, smoking, drinking, results from the logistic regression analyses revealed that those individuals who were carrying GG or GT genotype of rs2298423 were expected to face an increased risk for total cholesterol and low density lipoprotein cholesterol level than those individuals that carrying TT genotype in Uighur populations (OR = 3.140, 95% CI: 1.270-7.764, P = 0.013 in TC level; OR = 3.818, 95% CI: 1.761-8.280, P = 0.001 in LDL-C level). 4) The T-G haplotype appeared more frequent in the Uighurs while the T-A haplotype was more commonly seen in the Han population, respectively (both P < 0.001).</p><p><b>CONCLUSION</b>The mutational frequencies of the tagging SNPs in rs2298423 and rs6986132 loci of the ADRβ 3 gene presented obvious differences between Han and Uighur populations of Xinjiang. G allele carriers of rs2298423 seemed to face an increased risk for TC and LDL-C level in the Uighur populations in Xinjiang.</p>

Modified primary culture of neonatal mouse myocardial cells / 中国组织工程研究

BACKGROUND:A lot of work has been carried out on the development of the primary cultured rat myocardial cel s at home and abroad. The primary culture technology of rat myocardial cel s becomes more mature, but myocardial cel s from neonatal mice are not easy to be obtained under the same experimental conditions. The mouse genome has more similarities with the human genome, which has a higher research value. OBJECTIVE:To improve the primary culture method of neonatal mouse myocardial cel s, and to obtain myocardial cel s with high purity, vitality and original structure and function. METHODS:The mouse cardiac tissues were treated using an enzyme digestion method to isolate isolated single myocardial cel s:first, the cardiac tissues were digested using trypsin, and then col agenous fibers were treated with col agenase to isolate single myocardial cel s. The concentration and action time of trypsin and type II col agenase were adjusted, and the pH values of reagents and temperature of each step were strictly control ed. RESULTS AND CONCLUSION:At 24 hours after inoculation, the myocardial cel s began to be adherent;at 48 hours, independent pulsation of myocardial cel s could be observed;at 72 hours, myocardial cel s were cross-linked;and at 96 hours, myocardial cel s formed cel clusters and presented with consistent beating. The survival rate and purity of myocardial cel s were both over 95%. This modified method could successful y culture myocardial cel s with high purity and viablility from neonatal mice, and the structure and function of myocardial cel s could be retained. Therefore, it is a feasible culture method.

Association between the rs1007888 polymorphism of macrophage migration inhibitory factor gene and coronary heart disease in the Kazakhs of China / 中国组织工程研究

BACKGROUND:Macrophage migration inhibitory factor (MIF) is a multi-potent cytokine that makes considerable contribution to the regulation of inflammatory response and immune response in the body. MIF rs1007888 is associated with various inflammatory diseases, but the correlation between rs1007888 and coronary heart disease in the Kazakhs of China has been rarely explored. OBJECTIVE:To investigate the relationship between rs1007888 gene polymorphisms in MIF gene and coronary heart disease in the Kazakhs from Xinjiang Uygur Autonomous Region, China. METHODS:A total of 230 Kazakh patients with coronary heart disease evidenced by coronary arteriography between December 2012 and July 2014 were recruited, and another 478 Kazak controls were free from coronary artery disease with normal angiograms. Real-time fluorescence quantitative PCR assay was used to detect the rs1007888 polymorphisms of MIF gene. Alele and genotype distributions of the rs1007888 polymorphism were compared between patients and controls. RESULTS AND CONCLUSION:Distribution of genotypes in the two groups appeared to be in Hardy-Weinberg equilibrium (P> 0.05). The alele frequencies and genotypes of MIF-rs1007888 showed no significant difference between the two groups (P > 0.05). Therefore, the genetic variation of rs1007888 in MIF gene is not associated with coronary heart disease in the Kazakhs of China.

Recombinant adeno-associated virus serotype 9 transfection of atherosclerosis mice:determination of the optimal expression time in vivo / 中国组织工程研究

BACKGROUND:Recombinant adeno-associated virus serotype 9 has a high affinity in myocardial tissue, and the expression of recombinant adeno-associated virus serotype 9-enhanced green fluorescent protein (rAAV9-eGFP) in the aorta of atherosclerosis mice is not clear. OBJECTIVE:To explore the optimal time point of rAAV9-eGFP expression in the aorta of atherosclerosis mice. METHODS:Atherosclerosis model was established with high-fat diet in 30 ApoE-/-mice for 16 weeks. Among them, 25 mice were injected with 5.0×1011 vg (virus genomes) rAAV9-eGFP through the tail vein, while the remaining 5 mice were injected with saline, serving as the control group. The virus-transfected mice were kil ed at 14, 21, 28, 35 and 60 days after transfection, and aortic tissue was harvested. The expression of enhanced green fluorescent protein was detected with laser scanning confocal microscope. Western blot assays were used to detect the expression of enhanced green fluorescent protein in aorta. The expression of enhanced green fluorescent protein in vivo was observed and the optimal expression time point was determined. RESULTS AND CONCLUSION:rAAV9-eGFP effectively transfected the aorta of atherosclerosis mice, enhanced green fluorescent protein was expressed in aortic tissue, and the expression intensity increased gradual y with the increasing transfection time. The highest expression level was found at 35 days after transfection and then maintained stable at 60 days. There were significant differences at different time points after transfection (P<0.001). These data indicate that rAAV9-eGFP can be effectively expressed in the aorta of atherosclerosis ApoE-/-mice and rAAV9-eGFP can be regarded as the optimal vector in the treatment of atherosclerosis.

Combination of rat tail collagen and platelet-derived growth factor-BB for culture of human umbilical vein endothelial cells / 中国组织工程研究

BACKGROUND:Rat tail colagen type I can promote the increase of muscle fiber cels and migration and tube formation of endothelial cels, which is speculated to provide a more suitable internal environment for the growth of cels. OBJECTIVE:To observe the effect and safety of platelet-derived growth factor-BB (PDGF-BB) with rat rail colagen against apoptosis of human umbilical vein endothelial celsin vitro. METHODS:The passage 4 human umbilical vein endothelial cels were cultured in the medium of rat rail colagen and the reduction ratio in different time points was detected by Alamar Blue. The passage 4 human umbilical vein endothelial cels were divided into four groups and cultured in 24-wel culture plates: control group, PDGF-BB group, H2O2 group, PDGF-BB+colagen group. H2O2 was used to induce cel apoptosis in al the groups. Western blot was used to detect the expression of PDGF-BB, apoptosis-related protein and anti-apoptosis protein after 72 hours. Meanwhile, TUNEL method was used to detect cel apoptotic rate. RESULTS AND CONCLUSION:The tube formation in the human umbilical vein endothelial cels was more than that cultured in normal medium (P < 0.05). Cels cultured with rat tail colagen showed similar growth to normal control cels, indicating rat tail colagen had no cytotoxicity. The expressions of PDGF-BB, Bcl-2, and p-Akt in the PDGF-BB+colagen group were significantly higher than those in the other three groups (P < 0.05), but the expression of Bax was lower than that in the other three groups (P < 0.05). The apoptotic rate in the PDGF-BB+colagen group was lower than the PDGF-BB group and H2O2 group (P < 0.05). These findings indicate that rat tail colagen has no cytotoxicity to human umbilical vein endothelial cells, and rat tail collagen combined with PDGF-BB can predominantly enhance anti-apoptosis effects.

Cultured cardiomyocytes identificaiton and different methods of extractingβ3-AR membrane protein comparison / 天津医药

Objective To optimize primary cultures techniques of isolating neonatal rat cardiomyocytes and to com?pare three different methods of extractingβ3-adrenergic receptor(β3-AR)membrane protein from cultured neonatal rat car?diomyocytes. Methods TypeⅡcollagen and differential velocity adhesion were used to collect primary cardiomyocytes. To?tal protein method, ultracentrifugation method, extract kit method were used to isolate cardiomyocytesβ3-AR membrane pro?teins. The BCA method was applied for protein quantification. Relative content ofβ3-AR membrane protein and GADPH in the sample were examined by western blot. Results Optimizing culture and isolation skills can produce a great quantity of cardiomyocytes in high concentration.The kit method acquired a higher level of protein concentration(8.26±0.29)g/L than to?tal protein method(5.12±0.47)g/L does than ultracentrifugation method(3.20±0.37)g/L does all of which were with signifi?cant difference(P < 0.05). The concentration of β3-AR membrane protein was higher if obtained by kit method(0.22 ± 0.05)than ultracentrifugation method(0.09 ± 0.03)than total protein method (0.01 ± 0.01) with significant difference(P <0.05). Conclusion optimizing methodology can obtain abundant myocardial cells in high concentraion. The kit method of isolating primary culturedβ3-AR membrane proteins result in improved concentration and specificity of membrane protein.

Serum lipid levels and pathological observation of apolipoprotein E knockout mice with atherosclerosis at different weeks of age / 中国组织工程研究

BACKGROUND:The formation of atherosclerotic lesions in apolipoprotein E knockout mice is similar to that of human systemic atherosclerosis, and apolipoprotein E knockout mice are ideal animals for current establishment of atherosclerosis models. OBJECTIVE:To research the pathological process of atherosclerosis in apolipoprotein E knockout mice aged different weeks, and to explore the effect of different diets on the occurrence and development of atherosclerosis in apolipoprotein E knockout mice. METHODS:Male apolipoprotein E knockout mice aged 8 weeks old were randomly divided into two groups, and fed with high fat diet and normal diet, respectively, for 8, 12, 16, 20, and 24 weeks. RESULTS AND CONCLUSION:Serological detection revealed that serum total cholesterol, triglycerides and low density lipoprotein levels were significantly higher in different weeks of mice of high fat diet group than in the normal diet group (P<0.05), in a time-dependent manner. Gross and frozen oil red O staining showed that atherosclerotic plaque area of lumen was significantly larger in the high fat diet group than in the normal diet group (P<0.05), in a time-dependent manner. At this time, significant differences in plaque area of lumen at each week were detected between both groups (P<0.05). Apparent lipid plaque was visible in aorta at 16 weeks of high fat diet in mice. Results demonstrated that apolipoprotein E knockout mice of atherosclerosis were successful y established. The formation of lipid streaks and fiber hyperplasia was faster in high fat diet group than in the normal diet group.

Association between the level of fasting blood glucose over 35-year-old and carotid intima-media thickness in Han, Uygur and Kazak population from Xinjiang Uygur Autonomous Region from 2007 to 2010 / 中华预防医学杂志

<p><b>OBJECTIVE</b>To analyze the relationship between different levels of fasting blood glucose over 35-year old and carotid intima-media thickness (IMT) in Han, Uygur and Kazak adult population from Xinjiang Uygur Autonomous Region.</p><p><b>METHODS</b>From October 2007 to April 2010, the present study was performed in 13 935 inhabitants among Han, Uygur and Kazak adult population of aged 35 years old and over by multi-stage stratified cluster random sampling principles from 7 regions in Xinjiang Uygur Autonomous and we excluded the IMT over 0.9 millimeter, long-term out and the floating population. All subjects were measured fasting blood glucose and IMT values of carotid artery. The subjects were divided into three groups according to different fasting blood glucose levels: normal, impaired fasting glucose (IFG) and diabetes mellitus (DM) and we used the analysis of variance to compare the differences among groups of IMT. Multiple linear regression model was used to explore factors of carotid IMT.</p><p><b>RESULTS</b>The IMT of males of Han, Uygur and Kazak were (0.81 ± 0.29), (0.71 ± 0.27) and (0.79 ± 0.21) mm respectively, the differences were significant (F = 88.50, P < 0.05) . The IMT in DM group ((0.82 ± 0.29) mm) was significantly higher than the normal ((0.77 ± 0.26) mm) and the IFG groups ((0.79 ± 0.27) mm) (F = 7.49, P < 0.05). The IMT of females of Han, Uygur and Kazak were (0.72 ± 0.27), (0.63 ± 0.25) and (0.77 ± 0.22) mm, respectively, the differences were significant (F = 173.93, P < 0.05) . The IMT in DM group ((0.75 ± 0.29) mm) and the IFG groups ((0.74 ± 0.26) mm) were significantly higher than the normal group ((0.70 ± 0.25) mm) (F = 10.46, P < 0.05). Multivariate regression analysis showed that diastolic blood pressure (β = 0.101, P < 0.01) , total cholesterol (β = 0.056, P < 0.05) and fasting blood glucose (β = 0.023, P = 0.009) were independent risk factors of IMT.</p><p><b>CONCLUSION</b>The level of fasting blood glucose was an independent influence factor of carotid IMT and had a positive correlation in Han, Uygur and Kazak population of Xinjiang Autonomous Region.</p>

Study on the appropriate cutoff points of waist-hip-ratio for Kazakh people over 35 years of age, in Xinjiang / 中华流行病学杂志

<p><b>OBJECTIVE</b>The aim of this study was to identify the appropriate cutoffs for waist-hip-ratio (WHR) for Kazakh adults at high risk of cardiovascular diseases in Xinjiang.</p><p><b>METHODS</b>A Cardiovascular Risk Survey (CRS) study was carried out from October 2007 to March 2010, with 14 618 representative participants selected, including 4 094 Kazakhs. Complete data on 4 004 participants were gathered. The age span of the participants was from 35 to 88 years old with the mean age as 48.60 years. Anthropometric data, blood pressure, serum concentration of serum total cholesterol, triglyceride, low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C) and fasting glucose were documented. Prevalence rate, sensitivity, specificity and distance on the receiver operating characteristic (ROC) curve of each WHR values were calculated.</p><p><b>RESULTS</b>1) The prevalence rates of hypertension, hypertriglyceridemia were high, also with higher WHR seen in both men and women. 2) The prevalence of hypercholesterolemia was high with higher WHR seen in women. 3) The shortest distances in the receiver operating characteristic curves for hypertension, dyslipidemia, diabetes, or ≥ 2 of these risk factors suggested that the WHR cutoffs were 0.92 or 0.91 for men and 0.86 or 0.85 for women.</p><p><b>CONCLUSION</b>Higher cutoff for WHR was needed in the identification of patients over 35 at high risk of cardiovascular disease among Kazakh population.</p>

Incidence and distribution of chronic heart failure in the elderly from Xinjiang / 中华流行病学杂志

<p><b>OBJECTIVE</b>To investigate the prevalence and distribution of chronic heart failure (CHF) in the aged population of Xinjiang.</p><p><b>METHODS</b>Four-stage random sampling method was employed in this cross-sectional study to analyze the prevalence, risk factors and combined cardiovascular diseases of heart failure among different ethnic groups in aged (≥60 years and over) population of Xinjiang. Sample of studied population was recruited from 6 different regions in Xinjiang, namely Urumqi, Karamay, Fukang, Turfan Basin, Hetian and Ili Kazakh Autonomous Prefectures.</p><p><b>RESULTS</b>3 858 participants were surveyed, with the response rate as 89.14% . The prevalence of CHF was 4.30% in this cohort, including 2.74% in Han, 5.25% in Uighur and 6.32% in Kazakh ethnic groups respectively. The prevalence rates of CHF in different ethnic groups were significant different, statistically (χ(2) = 22.62, P = 0.00). Prevalence in males was 5.50%, predominant in the CHF, with females as 3.13%, and the difference between genders was significant (χ(2) = 13.65, P = 0.00). The prevalence rates of CHF increased in proportion with aging and were 3.39%, 3.68%, 5.12% and 6.82% in the 60-64, 65-69, 70-74, 75 years and over age groups, respectively. The prevalence rates of different age groups showed significant differences (χ(2) = 13.29, P = 0.004), and increased with age. The prevalence of CHF presented a rising trend (χ(2) = 12.07, P = 0.001).</p><p><b>RESULTS</b>of this study showed that atrial fibrillation was an independent risk factor for CHF (OR = 5.20, 95%CI: 2.32-11.70 and OR = 5.54, 95% CI:3.83-8.02). The most common combined single cardiovascular disease appeared to be hypertension(50 cases, the constituent ratio was 30.12%), followed by coronary heart disease (12 cases, 7.23%).</p><p><b>CONCLUSION</b>The prevalence of heart failure in population over 60 years was high in Xinjiang, and there showed ethnic differences. Hypertension and coronary heart disease were the basic cardiovascular diseases combined with heart failure in the various ethnic groups in Xinjiang. To strengthen epidemiological study on those high risk populations related to chronic heart failure was important on the strategies of prevention and treatment to this health problem.</p>

Detection rates of overweight and obesity in children and adolescents with different ethnicities in Xinjiang Uigur areas / 中华流行病学杂志

<p><b>OBJECTIVE</b>To investigate the detection rates of overweight and obesity among Han,Uygur and Kazakh Children and adolescents in Xinjiang.</p><p><b>METHODS</b>Random samples were used to analyze the detection rates on overweight and obesity based on data from Han,Uygur and Kazakh Children and adolescents aged 7-14 years from 3 regions-Hetian, Kashi and Fuhai prefectures in Xinjiang Uygur Autonomous Region. Anthropometric data including weight and height was collected. Overweight and obesity were defined according to body mass index [BMI, weight (kg)/height (m²)] cutoffs recommended by the Chinese Working Group on Obesity for Children (WGOC)aged 7-14 years.</p><p><b>RESULTS</b>The present study was performed in 11 894 Han, Uygur and Kazakh Children and adolescents aged 7-14 years. The overall prevalence rates of overweight and obesity were 5.6% and 2.9% respectively, which the difference between male and female was significant (χ² = 39.765, P < 0.05). Different ethnic students of overweight and obesity rate showed differences, with detection rate of overweight among Kazakh students was 12.0% , higher than the other ethnic groups. The obesity detection rate was 7.2% among Han students, which was the highest of the three ethnic groups, with differences statistically significant(χ² = 363.885, P < 0.05). Prevalence rates of both overweight and obesity decreased with increasing age in various ethnic groups, showing a clear downward trend, with the peak at 7-9 year-olds and the difference was statistically significant(χ² = 101.479, P < 0.05).</p><p><b>CONCLUSION</b>Both detection rates on overweight and obesity among children and adolescents were significantly lower in Xinjiang Uygur Autonomous Region than the national average level, with differences also seen in different ethnic, gender and age groups.</p>

Association between the single nucleotide polymorphisms of human CD36 gene and acute coronary syndrome / 中华流行病学杂志

<p><b>OBJECTIVE</b>The aim of the present study was to assess the association between the polymorphisms of CD36 gene and acute coronary syndrome(ACS).</p><p><b>METHODS</b>Genotypes of CD36 single nucleotide polymorphisms were detected under PCR-RFLP in 522 patients with ACS and 1 215 controls.</p><p><b>RESULTS</b>For people under Han and Uygur ethnicities, the distribution of genotypes and allele of rs1722505 was significantly different between ACS and the controls(all P < 0.05). For Han population, the frequency of A allele of rs17154181 was significantly lower in ACS group than that in the control group(P = 0.034). Results from logistic regression analysis showed that the AA+AG genotype of rs1722505 was significantly higher in ACS patients than that in controls both between the Hans and the Uygurs(OR = 1.436, 95%CI:1.047-1.970, P = 0.025;OR = 1.589, 95%CI:1.009-2.473, P = 0.046, respectively). For Han people,AA+AG genotype of rs17154181 was significantly lower in ACS patients than that in controls(OR = 0.667, 95% CI:0.494-0.900, P = 0.008).</p><p><b>CONCLUSION</b>Data from the present study suggested that the CD36 gene might serve as a genetic marker of ACS in both Han and Uygur populations.</p>

An improved protocol for primary culture of cardiomyocyte from neonatal rat / 中华心血管病杂志

<p><b>OBJECTIVE</b>The present study describes an improved in vitro culture method for obtaining high purity, vital and fully functional cardiomyocytes from neonatal rat.</p><p><b>METHODS</b>After cutting ventricular tissue with improved method, ventricular tissues were digested with low concentrations of trypsin overnight at 4 °C, and then underwent collagenase II digestion. Thereafter, cardiomyocytes were purified by combined differential adhesion and chemical inhibition methods.</p><p><b>RESULTS</b>Adherent cardiomyocytes were seen at 12 h after culture, spontaneously beating cardiomyocytes were observed at 24 h after culture, crosslinked cardiomyocytes were found at 48 h after culture, adhesion clustered cardiomyocytes were seen at 72 h after culture, dense network formed from inter-connected was evidenced together with radial arranged cell clusters and cell pseudopodia 96 h the mutual contact woven into and formed radically ordering cell clusters and island-like beating cardiomyocytes at 96 h after culture. The cell survival rate and purity were more than 98%.</p><p><b>CONCLUSION</b>Fully functional spontaneous beating cardiomyocytes can be obtained by the use of this improved primary neonatal rat cardiomyocytes culture method.</p>

Myocardial revascularization after myocardial infarction using endothelial progenitor cells combined with fibrin gel / 中国组织工程研究

BACKGROUND:Studies have shown that fibrin glue can promote the survival of myoblast grafts, reduce infarct size and induce neovascularization of infarct zone. OBJECTIVE:To understand the condition of revascularization of infarcted heart muscle using endothelial progenitor cells combined with degradable fibrin glue materials. METHODS:A total of 27 Sprague-Dawley rats were randomized into three groups, 9 rats in each group:non-myocardial infarction group, immediate transplantation group and 1-week post-infarction transplantation group. Then, these three groups were sub-grouped into two groups, respectively:endothelial progenitor cells+fibrin glue group (experimental group) and fibrin glue group (control group). At 3 and 8 weeks after transplantation, the rats were sacrificed in each group. The revascularization and function of infracted heart muscle were observed by microscope, immunohistochemistry and echocardiography. RESULTS AND CONCLUSION:Under the microscope, there were some lax connective tissues between the heart and chest in the experimental groups, but no difference existed between the experimental and control groups. The heart structure was normal relatively and difficult to be distinguished between the experimental and control groups histological y and immunological y, and there was no angeioma, vascular malformation and tumor. The number of revascularization of heart muscle showed no difference between experimental and control groups as wel as between different experimental groups. Additional y, there was no significant difference in cardiac function between experimental and control groups. Although there are no positive results of endothelial progenitor cells, we wil modify and improve the strategy and believe that the celldelivery system is of benefit and efficacy.

Location and expression of cyclin A2 in neonatal mouse myocardium / 中国组织工程研究

BACKGROUND:Cyclin A2 is a key regulator of cellcycle. Location and expression of cyclin A2 in neonatal mouse myocardium is not clear. OBJECTIVE:To observe the location and expression of cyclin A2 in neonatal mouse cardiomyocytes and its relationship with the exit of cardiomyocytes from cellcycle. METHODS:Neonatal mice were kil ed to take myocardial tissues at 0, 3, 7, 14 and 28 days after birth. Western blot were used to detect the expression of cyclin A2, proliferating cellnucleus antigen and Phospho-histone H3. Immunohistochemitry detection was used to detect the location of cyclin A2 and expression of proliferation cellnucleus antigen at different time after birth. RESULTS AND CONCLUSION:Western blot showed the decrease of cyclin A2 after birth til disappeared at day 4 (P=0.001). Cyclin A2 located mainly in the nucleus after birth and exported to the cytoplasm at day 14, and basical y disappeared at day 28. Proliferating cellnucleus antigen showed gradual y decreased tendency after birth. Mitosis specific marker, Phospho-histone H3, exhibited a gradual decrease after birth, which was consistent with cyclin A2 in expression intensity.

Platelet-derived growth factor-B gene transfection reduces ischemia and hypoxia-induced myocardial apoptosis / 中国组织工程研究

BACKGROUND:Platelet-derived growth factor-B (PDGF-B) is an effective pro-angiogenic growth factor, and adeno-associated virus type 9 (rAAV9) has a strong cardiomyocyte targeting affinity, which is an ideal vehicle for ischemic heart disease gene therapy. OBJECTIVE:To explore the PDGF-B gene transfection of in vitro neonatal rat myocardial cells mediated by rAAV9 against ischemia and hypoxia-induced cardiomyocytes apoptosis. METHODRat neonatal myocardial cells were isolated and cultured, and then transfected by rAAV9-PDGF-B and empty virus, rAAV9 with enhance green fluorescent protein (eGFP), under multiplicity of infection (MOI) of 105, 106 and 107, respectively. We observed the expression of eGFP under fluorescence microscopy every day, and used flow cytometry to measure transfection efficiency of vector rAAV9. Western blot and immunofluorescence were used to evaluate protein expression of PDGF-B. Myocardial ischemia and hypoxia injury model was established in vitro on the 5th day of transfection of rAAV9-eGFP and rAAV9-PDGF-B with 107 MOI. The number of myocardial apoptosis was measured by TUNEL assay. Western blot was employed to detect the protein expression of Bax and Caspase-3 which were related apoptosis, and the effect and its possible mechanism of PDGF-B gene overexpression against myocardial apoptosis were explored. RESULTS AND CONCLUSION:rAAV9 vector can efficiently transfect neonatal rat myocardial cells. eGFP and PDGF-B protein expressed in myocardial cells correctly and efficiently, and the expression intensity increased gradual y with the increasing of time course and MOI. The expression became stable on the 5th day, and the transfection efficiency showed significant difference among these groups (P<0.01). Myocardial apoptosis rate was significantly reduced in the rAAV9-PDGF-B group than the rAAV9-eGFP group (P<0.05), and protein levels of Bax and Caspase-3 in the rAAV9-PDGF-B group were significantly lower than those of the rAAV9-eGFP group (P<0.05). These data indicate that overexpression of PDGF-B gene can effectively reduce ischemia and hypoxia-induced myocardial apoptosis, and the possible mechanism might be by inhibiting Bax and Caspase-3 protein expression, which can provide evidence of rAAV9-PDGF-B vector in the gene therapy of ischemic heart diseases.

Recombinant adeno-associated virus serotype 9 with p65 ribozyme protects H9c2 cells from oxidative stress through inhibiting NF-κB signaling pathway / 老年心脏病学杂志（英文版）

Background Oxidative stress is a major mechanism underlying the pathogenesis of cardiovascular disease. It can trigger inflammatory cascades which are primarily mediated via nuclear factor-κB (NF-κB). The NF-κB transcription factor family includes several subunits (p50, p52, p65, c-Rel, and Rel B) that respond to myocardial ischemia. It has been proved that persistent myocyte NF-κB p65 activation in heart failure exacerbates cardiac remodeling. Mechods A recombinant adeno-associated virus serotype 9 carrying enhanced green fluorescent protein and anti-NF-κB p65 ribozyme (AAV9-R65-CMV-eGFP) was constructed. The cells were assessed by MTT assay, Annexin V–propidium iodide dual staining to study apoptosis. The expression of P65 and P50 were assessed by Western blot to investigate the under-lying molecular mechanisms. Results After stimulation with H2O2 for 6 h, H9c2 cells viability decreased significantly, a large fraction of cells underwent apoptosis. We observed a rescue of H9c2 cells from H2O2-induced apoptosis in pretreatment with AAV9-R65-CMV-eGFP. Moreover, AAV9-R65-CMV-eGFP decreased H2O2-induced P65 expression. Conclusions AAV9-R65-CMV-eGFP protects H9c2 cells from oxidative stress induced apoptosis through down-regulation of P65 expression. These observations indicate that AAV9-R65-CMV-eGFP has the potential to exert cardioprotective effects against oxidative stress, which might be of great importance to clinical efficacy for cardiovascular disease.

Comparative study of in vitro transfection of R65 ribozyme gene to different human vascular cells mediated by recombinant adeno-associated virus 9 / 中华微生物学和免疫学杂志

Objective To evaluate in vitro transfection of anti-nuclear factor-κB ( NF-κB) ribozyme gene to human umbilical vein endothelial cells (HUVECs) and human aortic smooth muscle cells (HASMCs) mediated by recombinant adeno-associated virus 9 carrying enhanced green fluorescent protein gene ( rAAV9-EGFP-R65 ) and to study their effects on cell proliferation and NF-κB P65 expression.Methods HUVECs and HASMCs were respectively transfected with rAAV9-EGFP-R65 at different multiplicity of infection ( MOI=1 ×105 , 1 ×106 and 1×107).The expression of EGFP was observed with fluorescence microscopy .Flow cytometry was performed to evaluate the transfection efficiency .Alamar Blue assay was used to measure the proliferation of the transfected cells.Western blot was used to detect NF-κB P65 expression .Results The fluorescence intensity was enhanced along with an increased MOI and an extended time of transfection .HUVECs and HASMCs transfected with rAAV 9-EGFP-R65 began to express EGFP at 24 h after transfection .The expression peak appeared on the sixth day in HUVECs, and the fifth day in HASMCs.The efficiencies of transfection in HUVECs at MOI of 1×105, 1×106 and 1×107 on the sixth day were (1.40±1.20)%, (12.30±1.35)%and (52.80±2.05)%, respectively.The trans-fection efficiencies of HASMCs on the fifth day were (5.30±1.04)%, (18.30±2.24)% and (52.40±3.21)%at MOI of 1×105 , 1×106 and 1×107 .Cell growth and morphology were not affected by transfection .Alamar Blue assay confirmed that there was no significant difference in the absorbance value between the transfected cells and two types of control cells .Western blot assay showed that the expression of NF-κB P65 was decreased by the trans-fection of rAAV9-EGFP-R65 in HUVECs and HASMCs .Conclusion rAAV9-EGFP-R65 can be efficiently trans-fected into two types of human vascular cells .It shows no inhibitory effects on cell proliferation , but can repress NF-κB P65 expression.