Red blood cell distribution width combined with lipoprotein-associated phospholipase A2 detection for improving diagnostic accuracy of coronary artery stenosis in patients with coronary artery disease / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the association of red blood cell distribution width (RDW) and lipoprotein-associated phospholipase A2 (LP-PLA2) with the degree of coronary artery stenosis in patients with coronary artery disease (CAD) and the value of RDW combined with LP-PLA2 detection in accurate evaluation of coronary artery stenosis.</p><p><b>METHODS</b>A total of 224 patients including 119 non-CAD cases and 105 CAD cases admitted in our hospital between June, 2013 and June, 2014 were enrolled in this study. The patients' baseline clinical data were collected and venous blood samples were obtained for detecting WBC, RDW-CV and LP-PLA2. The Gensini score of the CAD patients was calculated based on coronary angiographic findings.</p><p><b>RESULTS</b>Compared with the non-CAD patients, CAD patients had significantly higher RDW-CV (P=0.009) and LP-PLA2 (P=0.004) levels. The CAD patients with high Gensini scores had also significantly higher RDW-CV (P=0.001) and LP-PLA2 (P<0.001) levels than those with low scores; RDW-CV and LP-PLA2 were significantly correlated with the Gensini score, and the area under curve of their combined detection was 0.931.</p><p><b>CONCLUSION</b>Combination of RDW and LP-PLA2 can improve the diagnostic accuracy of the degree of coronary artery stenosis in patients with CAD.</p>

Change of IL-22R1 expression in human airway smooth muscle cells in response to different stimulating agents / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the effects of serum of asthmatic patients, dexamethasone, interleukin-4 (IL-4), interferon-gamma (IFN-γ) and transforming growth factor-β (TGF-β) on the expression of interleukin-22 receptor 1 (IL-22R1) mRNA and protein in HASMCs in vitro.</p><p><b>METHODS</b>IL-22R1 mRNA and protein expressions in HASMCs treated with different stimulating agents were measured by real-time PCR and Western blotting, respectively.</p><p><b>RESULTS</b>IL-22R1 mRNA and protein expressions in HASMCs were significantly increased after stimulation by serum from asthmatic patients, but decreased after co-stimulation with dexamethasone. IL-22R1 mRNA and protein expressions in the cells both increased after stimulation by IL-4, IFN-γ and TGF-β.</p><p><b>CONCLUSION</b>IL-22R1 in HASMCs might be involved in the pathogenesis of asthma, and the therapeutic effect of dexamethasone on asthma is mediated, at least partially, by IL-22R1. The effects of IFN-γ, IL-4, and TGF-β on asthma may also be attributed to their actions on HASMCs.</p>

Alteration of PTEN gene expression affects the migration of human airway smooth muscle cells in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the changes in human airway smooth muscle cell (HASMC) migration and related signaling pathway after interference with PTEN gene expression.</p><p><b>METHOD</b>HASMCs were infected with an adenovirus vector and RNA interference vector of human PTEN gene to establish the cell model with PTEN gene over-expression (Ad-GFP-PTEN-HASMC) and one with PTEN gene silencing (Ad-shPTEN-HASMC), using Ad-GFP-infected and a blank cells as the negative controls and LY294002 as the positive control. Fluorescence microscopy and flow cytometric analysis were used to evaluate the transfection efficiency, and Western blotting was performed to examine the expression of PTEN and the activation of AKT and ERK1/2 signal pathway. Transwell assay and wound healing assay were used to assess the migration of HASMCs.</p><p><b>RESULTS</b>The adenovirus over-expression vector and RNA interference vector significantly affected the expression of human PTEN gene. Up-regulation of PTEN gene resulted in a slow-down of the HASMC migration, an inhibition of PI3K/AKT signal pathway at the protein level but no changes in Ras-Raf-MEK1/2-ERK1/2 signal pathway. Down-regulated PTEN gene expression, however, was not associated with an enhancement of HASMC migration, but activated PI3K/AKT signal pathway and inhibited Ras-Raf-MEK1/2-ERK1/2 signal pathway.</p><p><b>CONCLUSION</b>Upregulation of PTEN gene can effectively inhibit airway smooth muscle cell migration, the effect of which is probably mediated by the PI3K/AKT pathway.</p>

Effect of physiological doses of testosterone on mitochondrial DNA deletion in castrated mice / 南方医科大学学报

<p><b>OBJECTIVE</b>To evaluate the effect of physiological doses of testosterone on mitochondrial DNA (mtDNA) deletion in the aortic vascular wall of castrated C57BL/6J mice.</p><p><b>METHOD</b>Twenty-four male C57BL/6J mice were randomized into normal control group (n=8), castrated+placebo group (castrated group, n=8), and castrated+physiological doses (1 mg/kg every 3 days) of testosterone group (n=8). The mice were fed normally for 3 months along with 8 mice with natural aging (18 months old), after which blood samples were obtained from all the groups for measurement of testosterone concentrations. The aortic mtDNA was extracted to analyze the deleted fragments using nested PCR, and fragments with deletions were purified and identified by sequence analysis.</p><p><b>RESULTS</b>Compared with the normal control group, the castrated group showed a significantly higher optical density ratio of the deletions [(18.1713 ∓ 2.4317)% vs (36.8475 ∓ 3.3365)%], but no significant difference was found between the castrated and natural ageing group [(42.3075 ∓ 3.6556)%]. The castrated+testosterone showed a lowered optical density ratio of (23.6488 ∓ 2.7634)% as compared with the castrated and natural ageing group, but a similar one with the normal control group. Sequence analysis identified 4 different types of deletions in the aging aorta at 3713, 3864, 4236, and 4415 bp, and the presence of direct repeats was confirmed to flank the deletions.</p><p><b>CONCLUSIONS</b>Multiple mtDNA deletions occur in ageing mice at a higher rate than in young mice. Testosterone deficiency is associated with increased aortic mtDNA deletions, which can be decreased by physiological doses of testosterone.</p>

Activation of Rho-kinase pathway is involved in angiotensin II-induced contraction of human airway smooth muscle cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate of the regulatory effect of Rho-kinase pathway activation on angiotensin II (Ang II)-induced contraction of human airway smooth muscle cells (HASMCs) in vitro.</p><p><b>METHODS</b>Cultured primary HASMCs were divided into control group, AngII group, AngII + irbesartan group and AngII + Y-27632 group with corresponding treatment. AngII-induced contraction of HASMCs was evaluated using collagen gel lattices and observed morphologically using immunofluorescence assay. Western Blotting was significantly performed to examine the protein expression of Rho-kinase signal pathway.</p><p><b>RESULTS</b>AngII-induced HASMC contraction was inhibited by treatments with irbesartan and Y-27632 as shown by gel contraction assay (P<0.001). Y-27632 treatment produced a stronger inhibitory effect than irbesartan on the expression of phosphorylated moesin, a substrate of Rho kinase (P<0.05).</p><p><b>CONCLUSION</b>AngII induces the contraction of HASMCs partially as a result of activation of Rho-kinase pathway.</p>

Pravastatin inhibits the expression of syndecan-4 protein in tumor necrosis factor-alpha-induced rat vascular smooth muscle cells in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of pravastatin on the proliferation of rat vascular smooth muscle cells (VSMCs) and expression of syndecan-4 protein induced by tumor necrosis factor-alpha (TNF-alpha).</p><p><b>METHODS</b>VSMCs cultured in vitro were exposed to 20 ng/ml TNF-alpha, 10 micromol/ml pravastatin, 20 micromol/ml pravastatin, 10 micromol/ml pravastatin with 20 ng/ml TNF-alpha, or 20 micromol/ml pravastatin with 20 ng/ml TNF-alpha for 24 h. The proliferation of the VSMCs was determined by non-radioactive MTS/PMS assay and the expression of syndecan-4 protein was detected by Western blotting using anti-syndecan-4 antibody.</p><p><b>RESULTS</b>Compared to the control group, TNF-alpha at 20 ng/ml significantly stimulated the proliferation of rat VSMCs (P<0.05). Pravastatin alone produced no obvious effect on VSMCs growth (P>0.05), but significantly inhibited TNF-alpha-induced VSMC proliferation (P<0.05). The expression of syndecan-4 protein in the VSMCs was significantly enhanced by 20 ng/ml TNF-alpha (P<0.01). Pravastatin alone did not affect the expression of syndecan-4 protein (P>0.05), but significantly inhibited TNF-alpha-induced enhancement of syndecan-4 protein expression (P<0.01).</p><p><b>CONCLUSION</b>Pravastatin can inhibit the proliferation and syndean-4 protein expression in rat VSMCs induced by TNF-alpha in vitro.</p>

Effects of simvastatin on plasma SOD, MDA and 8-iso-PGF2α in patients with stable angina / 南方医科大学学报

To observe the effects of simvastatin on plasma superoxide dismutase (SOD), malonaldehyde (MDA) and 8-iso-prostaglandin F2α (8-iso-PGF2α) as well as uric acid (UA) and serum lipids in patients with stable angina. METHODS Eighty-five patients with stable angina were divided into 4 groups, including hyperlipemia treatment group (HLT), hyperlipemia control group (HLC), normolipemia treatment group (NLT), and normolipemia control group (NLC). All the patients received routine treatment according to the guideline of CHD treatment, and those in the treatment groups were given Simvastatin (40 mg) every night, whereas those in the control group received placebo for 3 months. Before and after the treatments, the levels of plasma 8-iso-PGF2α were measured by enzyme-linked immunosorbent assay, and the plasma levels of SOD and MDA were detected by colorimetric method. LDL, HDL, TC, TG, and UA were also measured biochemically. RESULTS Compared with the control group, both of the treatment groups showed significantly increased levels of SOD and decreased MDA, 8-iso-PGF2α, UA and plasma lipids after the treatments (P<0.05). CONCLUSION In patients with coronary heart disease, simvastatins can decrease plasma lipids, inhibit lipid peroxidations, and promote the clearance of free radicals, thereby alleviating the oxidative stress.

Effect of recombinant adenovirus carrying PTEN gene on the proliferation of human airway smooth muscle cells in vitro and the mechanisms / 南方医科大学学报

<p><b>OBJECTIVE</b>To observe the effect of exogenous phophatase and tensin homolog deleted on chromosone 10 (PTEN) gene transfer via recombinant adenoviruses on the proliferation of human airway smooth muscle cells (HASMCs) in vitro and investigate the possible mechanisms.</p><p><b>METHODS</b>With a recombinant adenovirus vector containing PTEN (Ad-PTEN) constructed using the pAdxsi system, PTEN gene was transiently transfected into HASMCs and the transfection efficiency was determined by fluorescence microscope. RT-PCR and Western blotting were performed to detect the expression of PTEN mRNA and protein in the infected cells. MTS/PMS assay was used to analyze the proliferation of HASMCs, and the cell cycle changes of the transfected cells were evaluated by flow cytometry with PI staining. The expression levels of Akt and p-A kt proteins were detected by Western blotting, and P21 mRNA expression determined by RT-PCR.</p><p><b>RESULTS</b>The recombinant adenovirus Ad-PTEN showed a wild-type PTEN gene transfer efficiency of 98% at the multiplicity of infection (MOI) of 100. RT-PCR and Western blotting showed that infection with the recombinant adenovirus resulted in PTEN overexpression in the HASMCs, causing also increased ratio of G(0)/G(1) cells and proliferation inhibition of the ASMCs. The overexpression of PTEN significantly decreased the expression level of p-Akt but increased P21 mRNA expression.</p><p><b>CONCLUSION</b>The recombinant adenovirus containing PTEN can be successfully transfected into HASMCs cultured in vitro, resulting in PTEN overexpression at both the mRNA and protein levels. PTEN overexpression can efficiently inhibit the proliferation of HASMCs possibly through the PI3K/PKB/AKt and P21 pathways.</p>

Effects of dexamethasone on intracellular expression of Th17 cytokine interleukin 17 in asthmatic mice / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the effects of dexamethasone on intracellular expression of Th17 cytokine interleukin 17 and the mechanisms in asthmatic mice.</p><p><b>METHODS</b>Experimental asthma was induced by ovalbumin (OVA) sensitization in 20 in female Balb/c mice with (dexamethasone group, n=10) or without dexamethasone treatment (model group, n=10), with another 10 serving as the control group. The levels of IL-17 in the bronchoalveolar lavage fluid (BALF) and serum of the mice were measured by enzyme-linked immunosorbent assay (ELISA), and the airway inflammation was evaluated by HE staining. The expressions of IL-17 and RORgammat mRNA were measured by reverse transcription-polymerase chain reaction (RT-PCR), and the expression of RORgammat protein was measured by immunohistochemical staining.</p><p><b>RESULTS</b>The levels of RORgammat and IL-17 mRNA and protein in the asthmatic model group were significantly higher than those in the control group (P<0.01), and the increased expressions of RORgammat and IL-17 mRNA and protein in the asthmatic mice were significantly reduced by dexamethasone treatment (P<0.05).</p><p><b>CONCLUSION</b>Dexamethasone can inhibit the release of IL-17 probably by inhibiting RORgammat expression and blocking Th17 differentiation in asthmatic mice.</p>

Construction of a recombinant adenovirus vector containing PTEN and its expression in airway smooth muscle cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct a recombinant adenovirus vector containing phosphatase and tensin homolog deleted on chromosome 10 (PTEN) using the pAdxsi system.</p><p><b>METHODS</b>PTEN cDNA from plasmid pcDNA3-PTEN was cloned into the shuttle plasmid pShuttle-GFP-CMV. The shuttle vector was transformed into competent DH5alpha strain with the vector pAdxsi to achieve the homologous recombination. The recombinant construct was subsequently linearized with PacI and transfected into HEK293 cells via Lipofectamine 2000. The recombinant adenovirus particles were collected, and after titration, the recombinant adenovirus was traced by monitoring GFP expression under fluorescence microscope. The expression of PTEN mRNA and protein in the recombinant adenovirus vector and airway smooth muscle cells were detected by PCR and Western blotting, respectively.</p><p><b>RESULTS</b>GFP was expressed in HEK293 cells infected by recombinant adenovirus, and the expression intensity increased gradually with the passage of time, with obvious cytopathic effect (CPE) noted in the cells. After 3 cycles of amplification, the titer of adenovirus containing PTEN reached an appropriate level. The viral titer of pAdxsi-GFP-PTEN was 2x10(10) pfu/ml, and PTEN mRNA expression was detected by PCR. The homologous protein expressed in the infected human airway smooth muscle cells significantly increased in comparison with that in the control cells.</p><p><b>CONCLUSION</b>The recombinant adenovirus containing PTEN is constructed successfully, which provides an experimental basis for studying the role of PTEN gene in asthma therapy.</p>

Association of ATP-binding cassette transporter A1 R219K polymorphism with atrial fibrillation / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the association of ATP binding cassette transporter A1 (ABCA1) gene R219K polymorphisms with atrial fibrillation (AF) in Chinese population.</p><p><b>METHODS</b>A total of 250 patients with AF and 250 control subjects were selected. Polymerase chain reaction-restricted fragments length polymorphism (PCR-RFLP) was used to determine the ABCA1 genotype, and the serum concentration of C-reactive protein (CRP) and high-density lipoprotein cholesterol (HDL-C) were measured in all the subjects.</p><p><b>RESULTS</b>The frequency of the RR , RK , KK , allele R , allele K genotype of ABCA1 in AF group and control group was 42.0%, 42.8%, 15.2%, 34.0%, 43.2% and 22.8%, 63.4%, 36.6%, 55.6%, 44.4%, respectively. The frequency of the KK genotype was significantly higher in the control group than in the case group (P=0.03), and the frequency of the allele K genotype was significantly different between the two groups (P=0.012). The serum CRP concentrations was significantly higher in AF group than in the control group (P=0.004), but serum HDL-C level showed no difference between the two groups. The serum CRP concentrations were significantly higher in patients with RR genotype than in those with KK genotype (P=0.013), and patients with RR genotype had significantly lower HDL-C level than those with RK and KK genotypes (P=0.009 and 0.027, respectively).</p><p><b>CONCLUSION</b>Patients with AF have elevated serum CRP level in comparison with healthy individuals, and the K allele of R219K polymorphism is an independent protective factor against AF.</p>

Effects of losartan on left ventricular hypertrophy and plasma transforming growth factor-beta1 in elderly patients with hypertension / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effects of losartan on left ventricular hypertrophy (LVH) and plasma transforming growth factor-beta1 (TGF-beta1) in elderly patients with essential hypertension (EH).</p><p><b>METHODS</b>The elderly patients with EH were divided into two groups, namely EH+LVH group and EH group according to the data of echocardiogram. The systolic and diastolic blood pressures of the patients were monitored. Plasma TGF-beta1 was measured before and after 6 months' treatment with losartan, and the relationship between TGF-beta1 and other index were analyzed.</p><p><b>RESULTS</b>After 6 months' treatment, the blood pressure of EH+LVH group and EH group were significantly lowered (P<0.01). Significant improvement of IVSTd, LVPWd, E/A, and LVMI (P<0.01) and obvious reduction of plasma TGF-beta1 (P<0.01) occurred in EH+LVH group after 6 months' treatment. Correlation analyses indicated that the plasma TGF-beta1 level was positively correlated to LVMI (P<0.01).</p><p><b>CONCLUSION</b>Losartan can reversed LVH in elderly patients with EH partially by lowering plasma TGF-beta1 level.</p>

Role of antigen-specific T cell-mediated immune response in coronary heart disease / 南方医科大学学报

<p><b>UNLABELLED</b>To investigate the effect of immune response mediated by the T cells stimulated with the specific antigen (oxidized low-density lipoprotein, oxLDL) on plaque stability in coronary heart disease.</p><p><b>METHODS</b>This study involved 20 patients with acute myocardial infarction (AMI), 34 with unstable angina pectoris (UAP), 27 with stable angina pectoris (SAP) and 22 healthy control subjects. With MTS/PMS colorimetric assay, the T cells from all the subjects were tested for proliferative response to stimulation by 5 microg/ml oxLDL and 5 microg/ml low-density lipoprotein (LDL). Interferon-gamma (IFN-gamma) concentration in the proliferative response of the T cells was measured with enzyme-linked immunosorbent assay (ELISA).</p><p><b>RESULTS</b>The proliferative response of the T cells elicited by 5 microg/ml oxLDL stimulation was significantly higher in the AMI and UAP groups than in the SAP and control groups (P<0.05). Similarly, IFN-gamma concentration in the proliferative response of the T cells to 5 microg/ml oxLDL stimulation was significantly higher in the former two groups (P<0.05). In the AMI and UAP groups, 5 microg<ml oxLDL stimulation resulted in significantly higher IFN-gamma concentration in the proliferative response of the T cells than 5 microg/ml LDL stimulation (P<0.001).</p><p><b>CONCLUSION</b>The immune response mediated by the T cells to specific antigen stimulation, especially the immune response mediated by T helper type 1 (Th1) cells secreting IFN-gamma, may play an important role in the plaque instability and the occurrence of acute coronary syndrome.</p>

Shikonin inhibits the proliferation of human airway smooth muscle cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the inhibitory effect of shikonin on the proliferation of human airway smooth muscle cells (HASMCs) in vitro.</p><p><b>METHODS</b>HASMCs from the trachea were obtained by primary culture of the tissue explants and adherent culture. The HASMCs were exposed to shikonin at 0 (control group), 0.5, 1, 2, 5, 10, 20, and 40 micromol/L for 12, 24, and 48 h, after which the cell proliferation was assessed by 3-(4,5-carboxymethoxypheny1)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) colorimetric assay. Flow cytometry was used for cell cycle analysis of the HASMCs exposed to shikonin at 40, 20, 10, 5 micromol/L and 0 micromol/L (control group) for 24 h. Immunocytochemistry with SP method was performed to detect the expression of proliferating cell nuclear antigen (PCNA) in the HASMCs treated with shikonin at 20 micromol/L and 0 micromol/L (control group) for 24 h.</p><p><b>RESULTS</b>Shikonin at the concentrations of 20 and 40 micromol/L significantly inhibited the proliferation of the cells (P<0.05), and the effect was especially obvious after 48 h of cell exposure, with inhibition rates of 30.1% and 42.9%, respectively. No significant difference was found between the two concentrations for their cell growth inhibition effects (P>0.05). Shikonin at the concentrations of 20 and 40 micromol/L caused significant cell cycle arrest in G(0)/G(1) phase (P<0.05), the effect of which, however, was not concentration-dependent (P>0.05). Shikohin at 20 micromol/L significantly down-regulated the expression of PCNA in the cells (P<0.05).</p><p><b>CONCLUSION</b>Shikonin can inhibit the proliferation of HASMCs in vitro.</p>

Construction and identification of adenovirus vector for double-mutant human hypoxia-inducible factor-1alpha gene / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct an adenovirus vector containing the double-mutant hypoxia-inducible factor-1alpha (HIF-1alpha) gene for exploring the therapeutic angiogenesis for coronary heart disease.</p><p><b>METHODS</b>Human double-mutant HIF-1alpha cDNA was obtained from PCR of pShuttle-2-HIF-1alpha containing the mutant HIF-1alpha gene (564). The recombinant adenoviral plasmid containing mutant HIF-1alpha cDNA was constructed by ligation of recombinant pShuttle2 containing double-mutant HIF-1alpha cDNA and Adeno-X viral DNA, followed by its identification and transfection into adenoviral packaging cells HEK293 via lipofectamine 2000.</p><p><b>RESULT AND CONCLUSION</b>The recombinant pAdeno-HIF-1alpha was correctly constructed and verified by restriction endonuclease and DNA sequencing analyseis. This recombinant adenovirus containing the double-mutant HIF-1alpha gene may facilitate further investigation of mutant HIF-1alphagene therapy for coronary heart disease.</p>

Association between ADAM33 gene polymorphism and bronchial asthma in South China Han population / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the association between the polymorphism of T(1) locus allele in ADAM33 gene and bronchial asthma in South China Han population.</p><p><b>METHODS</b>Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and DNA sequencing were used to determine the polymorphism of T(1) locus allele in ADAM33 gene in 160 unrelated patients with asthma and 95 unrelated healthy controls from South China Han population.</p><p><b>RESULTS</b>No significant difference was found in T(1) locus allele distribution frequency in populations of UK, US, Germany, Korea, and South China (Chi(2)=9.085, P=0.109). The frequencies of the genotypes (TT, TC, CC) were 80.6% (n=129), 16.9% (n=27) and 2.5% (n=4) in the 160 asthmatic patients and 94.7% (n=90), 3.2% (n=3) and 2.1% (n= 2) in the 95 controls, respectively, showing a significant difference in the distribution of the genotypes (TT, TC, CC ) between asthmatic patients and healthy controls (Chi(2)=10.955, P<0.05). The frequencies of the alleles (T, C) were 0.891 and 0.109 in the asthmatic patients and 0.963 and 0.037 in the controls, respectively, showing also a significant difference in the allele frequency between them (Chi square =8.299, P<0.05). The presence of C allele of ADAM33 gene T1 locus was found to be a greater risk factor in asthmatic patients than in the healthy controls. The odds ratio (OR) of TC and TC+CC were 6.279 (1.849-21.328) and 4.326 (1.620-11.550), respectively, with P value of 0.001 and 0.002, respectively, in comparison with TT genotype.</p><p><b>CONCLUSION</b>The polymorphism of T(1) locus allele in ADAM33 gene is associated with the susceptibility to asthma in South China Han population.</p>

Tumor necrosis factor-alpha regulates the proliferation and syndecan-4 expression of human umbilical vein endothelial-like cells cultured in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effects of tumor necrosis factor-alpha(TNF-alpha) on syndecan-4 protein expression and proliferation of cultured human umbilical vein endothelial-like cells (HUVECs) in vitro.</p><p><b>METHODS</b>HUVECs exposed to different concentrations of TNF-alpha(100, 20, 10, and 1 ng/ml) were cultured for 24 h and 36 h to observe their proliferation in comparison with the control group. The cell proliferation rate was determined by non-radioactive MTS/PES assay. The expression of syndecan-4 protein was evaluated by immunoblotting technique using anti-syndecan-4 antibody. Results The proliferation rate of the endothelial-like cells was 1.956-/+0.214 in the control group, and 2.154-/+0.250, 2.260-/+0.151, 2.118-/+0.205 and 2.106-/+0.136 in TNF-alpha-treated groups corresponding to TNF-alpha concentrations of 100, 20, 10 and 1 ng/ml at 24 h, respectively. It was shown that TNF-alpha significantly stimulated cell proliferation at the concentration above 1 ng/ml (P<0.05) as compared with the control group (P<0.05). The proliferation rate of the endothelial-like cell was 1.915-/+0.236 in the control group, and 2.067-/+0.328, 2.207-/+0.150, 2.052-/+0.126 and 2.051-/+0.180 in TNF-alpha-treated groups corresponding to TNF-alphaconcentrations of 100, 20, 10 and 1 ng/ml at 36 h, respectively. The expression of syndecan-4 protein was significantly enhanced by TNF-alpha.</p><p><b>CONCLUSIONS</b>TNF-alpha can stimulate HUVEC proliferation, and expression of syndean-4 may represent an additional component of the pro-inflammatory, growth-stimulating pathways that are activated in response to changes in the vascular wall.</p>

Effects of furosemide, antisterone and hydrochlorothiazide on expression of kidney aquaporin-2 gene and urine aquaporin-2 excretion in rats / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate effects of the furosemide, antisterone and hydrochlorothiazide on expression of kidney aquaporin-2 (AQP(2)) gene and urine aquaporin-2 excretion in rats.</p><p><b>METHODS</b>Forty SD rats were randomized into 4 groups, namely the control group, furosemide group, antisterone group and hydrochlorothiazide group with corresponding treatment. Blood and urine samples were collected from the rats for measurement of serum Na(+), urine volume and urine osmolality during medication. Semi-quantitative RT-PCR was performed to measure kidney inner medullary AQP(2) and vasopressin V(2)-R mRNA. Western blotting was employed to detect kidney inner medullary AQP(2) protein expression. Urine AQP(2) concentration was measured by enzyme-linked immunosorbent assay (ELISA).</p><p><b>RESULT</b>Urine volume and urinary AQP(2) excretion were both increased in rats treated with the 3 drugs as compared with that of the control group. However, urine osmolality was lower in furosemide group but higher inhydrochlorothiazide and antisterone groups than in the control group (P<0.05). The kidney inner medullary AQP(2) mRNA, V(2)-R mRNA and AQP(2) protein expression of furosemide group increased in comparison with that of the control group (Plt;0.05). In hydrochlorothiazide group, however, the above parameters were all decreased (Plt;0.05).</p><p><b>CONCLUSION</b>The three classes of diuretics can all increase the excretion of the urinary AQP(2) but have different effects on the inner medullary AQP(2) mRNA and protein expression in normal rats. Hydrochlorothiazide reduces kidney AQP(2) mRNA and protein expression, while furosemide increased kidney AQP(2) gene expression.</p>

Sleep-disordered breathing and left ventricular remodeling in patients with chronic heart failure / 南方医科大学学报

<p><b>OBJECTIVE</b>The investigate the prevalence of sleep-disordered breathing (SDB) and evaluate its impact on left ventricular remodeling in adult patients with chronic heart failure (CHF).</p><p><b>METHODS</b>Ambulatory sleep recording for 8 h was performed using Embletta PDS (Medcare, Iceland) in 74 patients with CHF, and the left ventricular ejection fraction (LVEF), internal end-diastolic diameter (LVIDd) and left ventricular mass weight (LVMW) were measured using M-mode and two-dimensional echocardiography.</p><p><b>RESULTS</b>The incidence of SDB defined as an apnea-hypopnea index (AHI, namely the number of apnea-hypopnea events per hour during sleep) no less than 10 was 62.16% in these CHF patients (77.78% in male and 37.93% in female patients). Of the 74 patients 31.1% had mainly obstructive sleep apnea (OSA) and 17.6% had central sleep apnea (CSA). There was a moderate inverse correlation between LVEF and AHI (P=0.004, r=-0.366). LVIDd in patients with CHF and SDB was significantly greater than that in patients with isolated CHF (46.67+/-7.29 vs 55.70+/-11.87 mm, P=0.001). The left ventricular myocardial weight was also greater in patients with CHF and SDB than in patients with isolated CHF (208.58+/-64.19 vs 291.03+/-121.54, P=0.001).</p><p><b>CONCLUSION</b>Our results suggest a higher prevalence of SDB in patients with CHF than in general population, and the prevalence is even higher in patients with severe CHF in relation to left ventricular remodeling. SDB contributes to the progression of CHF and further cardiac decline by a vicious cycle.</p>

A novel polymorphism in the ATP-binding cassette transporter A1 gene (M233V) in Chinese patients with coronary artery disease / 中华心血管病杂志

<p><b>OBJECTIVE</b>To identify ATP-binding cassette transporter A1 (ABCA1) gene polymorphism in Chinese patients with coronary artery disease (CAD).</p><p><b>METHODS</b>Single Nucleotide Polymorphisms in the ABCA1 gene were detected by polymerase chain reaction (PCR)-single strand conformation polymorphism (SSCP)-DNA sequence and restriction-fragment length polymorphism (RFLP) method in 112 patients with CAD.</p><p><b>RESULTS</b>A novel polymorphism in the ABCA1 gene was found in two patients: M233V which exists in exon7 of ABCA1 gene and it's cDNA location is A1092G and converse 233 amino acid from Methionine to Valeric. We further collected the blood samples from 16 family members of one proband and M233V polymorphism was found in 5 out 16 family members.</p><p><b>CONCLUSION</b>M233V is a novel polymorphism in the ATP-binding cassette transporter A1 gene and this AG genotype had family proneness.</p>