[Peroxisome proliferator-activated receptor α/γ agonist tesaglitazar stabilizes atherosclerotic plaque in diabetic low density lipoprotein receptor knockout mice].

OBJECTIVE: To investigate the effects of peroxisome proliferator-activated receptor (PPAR) α/γ agonist on atherosclerotic plaque stabilization in diabetic LDL receptor knockout (LDLr-/-) mice. METHODS: Female 4-week-old LDLr-/- mice fed with high-glucose and high-fat diet for 4 weeks were randomly divided into three groups (n = 15 each): control group (only fed with high-glucose and high-fat diet), diabetic group [induced by high-glucose and high-fat diet combined with a low-dose of streptozotocin (STZ)] without tesaglitazar and with tesaglitazar (20 µg/kg oral treatment). After 6 weeks, the mice were sacrificed, body weight, fasting blood glucose (Glu), total cholesterol (TC), triglyceride (TG) levels were measured. The expression of ICAM-1, VCAM-1, MCP-1 in the brachiocephalic atherosclerotic lesions were determined by Western blot and immunohistochemistry, respectively. Brachiocephalic artery was prepared for morphologic study (HE, oil red O, Sirius red staining) and immunohistochemical analysis (macrophage surface molecule-3, α-smooth muscle actin), respectively. RESULTS: Serum TC [(32.34 ± 3.26) mmol/L vs. (16.17 ± 1.91) mmol/L], TG [(3.57 ± 0.99) mmol/L vs. (2.21 ± 0.11) mmol/L] and Glu [(15.21 ± 4.67) mmol/L vs. (6.89 ± 0.83) mmol/L] levels were significantly higher in diabetic group than in the control group (all P < 0.01). The expression of ICAM-1 (2.31 ± 0.35 vs.1.34 ± 0.21), VCAM-1 (1.65 ± 0.14 vs.0.82 ± 0.26), MCP-1 (2.27 ± 0.16 vs.1.56 ± 0.23) were significantly upregulated in diabetic group compared with control group (all P < 0.01). Brachiocephalic atherosclerotic plaque area [(4.597 ± 1.260)×10(3) µm(2) vs. (0.075 ± 0.030)×10(3) µm(2)], lipid deposition [(47.23 ± 2.64)% vs. (9.67 ± 1.75)%], Mac-3 positive area [(19.15 ± 3.51)% vs. (1.72 ± 0.16)%], α-smooth muscle actin [(5.54 ± 1.17)% vs. (2.13 ± 0.41)%] and collagen content [(4.27 ± 0.74)% vs. (0.43 ± 0.09)%] were all significantly larger/higher in diabetic LDLr-/- mice than in the control group (all P < 0.01). While tesaglitazar treatment significantly reduced serum TC [(30.47 ± 3.18) mmol/L], TG [(3.14 ± 0.71) mmol/L] and Glu [(7.92 ± 1.28) mmol/L] levels (all P < 0.01). Similarly, the expression of ICAM-1 [(1.84 ± 0.22)], VCAM-1 [(1.27 ± 0.11)], MCP-1 [(1.83 ± 0.24)], brachiocephalic atherosclerotic lesion area[(1.283 ± 0.410)×10(3) µm(2)], lipid deposition[(23.52 ± 1.39)%] were also significantly reduced by tesaglitazar (all P < 0.05). Moreover, tesaglitazar increased α-smooth muscle actin [(9.46 ± 1.47)%] and collagen content [(6.32 ± 1.15)%] in diabetic LDLr-/- mice (all P < 0.05). In addition, lipid deposition and Mac-3 positive areas [(10.67 ± 0.88)% vs. (15.83 ± 1.01)%] in the aortic root were also reduced in tesaglitazar treated diabetic LDLr-/- mice (P < 0.01). CONCLUSIONS: Tesaglitazar has anti-inflammatory effects in the diabetic LDLr-/- mice. Tesaglitazar could reduce lipid deposition, increase collagen and α-SMA content in the brachiocephalic atherosclerotic lesions, thus, stabilize atherosclerotic plaque in this model.

Serum total adiponectin level and risk of cardiovascular disease in Han Chinese populations: a meta-analysis of 17 case-control studies.

OBJECTIVE: To systematically evaluate low serum adiponectin level as a risk factor for cardiovascular disease (CVD). A meta-analysis was performed to quantitatively analyse the association of serum total adiponectin level with CVD using previous case-control studies in Han Chinese populations. METHODS: Several electronic databases were searched for relevant articles up to July 2011. A total of nine (n = 933), eight (n = 939) articles were included in each meta-analysis regarding the association of serum adiponectin level with coronary heart disease (CHD) and ischaemic stroke, respectively. Publication bias was examined by the Egger's linear regression test. Sensitivity analysis was performed by omitting one study at a time, and the pooled standardized mean difference (SMD) was estimated using fixed-effects model and random-effects model, respectively. RESULTS: Serum total adiponectin concentrations were lower in patients with CHD and ischaemic stroke, with pooled SMD of -1·41 (95% CI -1·69, -1·12, P < 0·00001) and -1·69 (95% CI -2·04, -1·33, P < 0·00001), respectively. By performing a meta-regression analysis, homeostasis model assessment of insulin resistance, study size, adiponectin measurement assays, gender and mean body mass index of cases failed to account for heterogeneity for comparisons between lower adiponectin level and ischaemic stroke. However, study size had significant effect on the association of lower adiponectin level with CHD and accounted for 96·72% of the between-study variance. No publication bias was detected. No single study was found to affect the overall result of each analysis by sensitivity testing. CONCLUSIONS: The accumulated evidence suggested that low serum adiponectin level increased the risk of a first cardiovascular event in the Han Chinese population. Further study is recommended with larger sample size to explore the role of hypoadiponectinemia in the causation of CVD.

Pegylated Interferon-α Plus Ribavirin Therapy Improves Left Ventricular Diastolic Dysfunction in Patients With Chronic Hepatitis C Attaining Sustained Virological Response.

BACKGROUND: Pegylated interferon (pegIFN) in combination with ribavirin (RBV) has successfully improved the rate of sustained virological response (SVR) in chronic hepatitis C virus (HCV) infected individuals, which reduces the progression of the chronic liver disease. However, the influence of combination therapy (pegIFN/RBV) on cardiac function has yielded ambiguous results. The present study aimed to evaluate the effects of combination therapy with pegIFN/RBV on cardiac function of HCV-infected individuals with SVR. MATERIALS AND METHODS: Cardiac function was assessed and correlated in 142 treatment-naïve patients with HCV infections by determining cardiac biomarkers and echocardiography before treatment and for 24 weeks post-treatment. RESULTS: An SVR was achieved by 50.7% of all patients. Serum N-terminal pro-B-type natriuretic peptide levels were significantly higher in all patients before treatment and decreased significantly 24 weeks post-treatment in the SVR group (62.84 [36.98-102.73] versus 22.87 [15.64-56.92] pg/mL, P < 0.01). Peak early diastolic annular velocity (E') was significantly lower (7.69 ± 2.48 versus 9.74 ± 2.68cm/s, P < 0.001) and E/E' was higher (10.04 ± 2.51 versus 8.18 ± 2.31, P < 0.001) in all patients with SVR. However, there were no statistically significant differences in biomarkers and echocardiographic parameters for patients without SVR. In addition, multivariate analysis identified age (odds ratio [OR] = 1.076; 95% CI: 1.031-1.125; P < 0.001), NT-proBNP (OR = 1.122; 95% CI: 1.002-1.248; P = 0.015), and SVR (OR = 0.532; 95% CI: 0.214-0.895; P = 0.023) as statistically significant independent variables associated with left ventricular diastolic dysfunction. CONCLUSIONS: The present study showed no adverse effects of combination therapy on cardiac function of HCV-infected individuals with SVR. Subsequent viral eradication resulted in improvement of left ventricular diastolic dysfunction.

[A novel bifunctional in vivo mutation detection system].

A shuttle plasmid pMCLacI/neo with two copies of LacI was integrated into mouse genome and a novel system which could detect in vivo mutation of both expression and silence genes was constructed, enabling the comparative analysis of their mutation spectra and mutant frequencies. 486 fertilized eggs from C57BL/6 mice with microinjected pMCLacI/neo plasmid were transferred into oviducts of 18 pseudo-pregnant mice, and 32 alive offsprings were screened and identified by using PCR and Southern blotting. Genomes of 5 mice had pMCLacI/neo plasmid integrated, as verified by Southern blot after the PCR screening. Only one of the two LacI in pMCLacI/neo was in expression state; and this established a model, that the status in vivo of both gene expression and silencing could be simulated. This kind of mice might be used as a novel bifunctional mutation detection system in vivo.

[A new strategy for detecting Lac I mutants based on pMCLac I/neo transgenic mice].

We studied a novel mutation detection method of the two Lac I target genes in pMCLac I/neo transgenic mice. The transgenic mice that contain two types of Lac I genes in pMCLac I/neo vector are different from the transgenic mice carrying only one target gene. Therefore a novel method to detect mutation quickly and efficiently has become a new project after the establishment of pMCLac I/neo transgenic mice. In this paper, a positive selection system--M9/L is used. The result showed that M9/L positive selection system was able to detect the two Lac I target genes, suggesting that it is a rapid and effective method to detect the target mutations in the pMCLac I/neo transgenic mouse.

[Establishment of a hemophilia B transgenic mouse model on the basis of coagulation factor IX gene knock-out mouse].

This study aimed to introduce a site specific point mutation into the human coagulation factor IX gene expressing vectors (pMe4bAIXml plasmid) for microinjection and to obtain transgenic mouse containing copies of a stably integrated pMe4bAIXml plasmid on the basis of coagulation factor IX gene knock-out mouse model as an more efficient animal model of hemophilia b. The site specific point mutation was introduced into pMe4bAIXml plasmid which consists of human coagulation factor IX gene including the entire coding sequence and a shortened first intron, four copies of the mouse MCK enhancer, chicken beta-actin promotor and poly A signal sequence. The vector was linearized and injected into 817 fertilized eggs of mice in which coagulation factor IX gene has been knocked out. The manipulated embryos were transferred into the oviducts of 45 pseudopregnant females, from which 63 offsprings were obtained. The genomic DNAs of these offsprings were analyzed with PCR and genomic Southern blotting. Six mice were found to carry copies of the intact pMe4bAIXml plasmid containing a point mutation and used as founders to establish transgenic mouse lineages.

Tesaglitazar ameliorates non-alcoholic fatty liver disease and atherosclerosis development in diabetic low-density lipoprotein receptor-deficient mice.

Previous research has demonstrated that the dual PPARα/γ agonist tesaglitazar reduces atherosclerosis in a mouse model of hyperlipidemia by reducing both lipid content and inflammation in the aorta. However, much of the underlying mechanism of tesaglitazar in non-alcoholic fatty liver disease (NAFLD) remains less clear. The aim of the present study was to determine whether tesaglitazar attenuates NAFLD and atherosclerosis development in diabetic low-density lipoprotein receptor-deficient (LDLr(-/-)) mice. Female LDLr(-/-) mice (3 weeks old) were induced by a high-fat diet (HFD) combined with low-dose streptozotocin (STZ) injection to develop an animal model of type 2 diabetes (T2DM). The mice were randomly divided into two groups: diabetic group (untreated diabetic mice, n=15) and tesaglitazar therapeutic group (n=15, 20 µg/kg/day oral treatment for 6 weeks). Fifteen LDLr(-/-) mice were fed with an HFD as the control group. Tesaglitazar decreased serum glucose and lipid levels compared with the diabetic mice. Tesaglitazar significantly reduced atherosclerotic lesions, lipid accumulation in the liver, macrophage infiltration, and decreased total hepatic cholesterol and triglyceride content compared to the diabetic mice. In addition, tesaglitazar reduced inflammatory markers at both the serum and mRNA levels. Our data suggest that tesaglitazar may be effective in preventing NAFLD and atherosclerosis in a pre-existing diabetic condition by regulating glucose and lipid metabolism, and the inflammatory response.

Comparative study of 4Fr catheters using the ACIST variable rate injector system versus 6Fr catheters using hand manifold in diagnostic coronary angiography via transradial approach.

BACKGROUND: The transradial approach is regarded as a useful vascular site for coronary procedures. The aim of this study was to test whether 4Fr catheters assisted by ACIST variable rate injector system can produce comparable angiographic quality and reduce the risk of radial artery injury compared to hand manifold 6 Fr catheters. METHODS: A total of 1816 patients were studied consecutively, among whom 856 patients received coronary angiography by 4 Fr catheters (4Fr group) and 960 patients by 6 Fr catheters (6Fr group). Angiographic and procedural characteristics were observed and recorded. The luminal inner radial arterial diameter before and after the procedure were collected. RESULTS: The baseline clinical characteristics were similar in both groups. There were no significant differences in procedure time, radiation dose and quality scores in both groups (P > 0.05), but more contrast media was delivered in the 6Fr group (P < 0.001). The mean radial arterial diameter six months after the procedure in the 6Fr group reduced significantly compared to that measured one day prior to the procedure (P < 0.001). CONCLUSIONS: Coronary angiography using the 4Fr catheters with Acist power injection system can achieve an acceptable diagnostic quality while at the same time minimizing radial artery injury and contrast media consumption.