[ABSTRACT]AIM:ToevaluatetheeffectsofantisenseTGF-β1oligodeoxynucleotide(ASTGF-β1)ontheex-pression of TGF-β1 , deposition of extracellular matrix ( ECM) and the neointima formation in the arteries after balloon inju-ry.METHODS:The unmodified and phosphorothioate-modified AS TGF-β1 which containing 15 bases and surrounding the initiation codon region (ATG) of rat TGF-β1 complementary DNA (cDNA) were designed.At the same time, sense TGF-β1 oligodeoxynucleotide ( S TGF-β1 ) with the base sequence complement to AS TGF-β1 was synthesized as a control . The oligodeoxynucleotides were introduced into in vivo and in vitro experiments , respectively .RESULTS:The AS TGF-β1 significantly inhibited the protein expression of TGF-β1 in a concentration-dependent manner , and S TGF-β1 did not have the same effect.Furthermore, no effect of the AS TGF-β1 on the mRNA expression of TGF-β1 in injured VSMCs was ob-served.Moreover, for the injured VSMCs, AS TGF-β1 significantly and concentration-dependently inhibited the basal DNA synthesis.Both AS TGF-β1 and S TGF-β1 did not exhibit dose-dependent effects on DNA synthesis in uninjured VSMCs . Fibronectin ( FN) mRNA expression in injured VSMCs was significantly decreased by AS TGF-β1 in a concentration (0.01~1 μmol/L)-dependent manner .AS TGF-β1 significantly increased the mRNA expression of contractile marker SM 22α, and decreased the mRNA expression of synthetic markers osteopontin and matrix Gla , especially at the concentration of 0.01μmol/L and 0.1 μmol/L.After treatment with AS TGF-β1 (90 μg· kg-1 · d-1 ) for 28 d, the neointima formation was significantly inhibited , and the area ratio of intima/media was markedly decreased by 68% compared with untreated group , but S TGF-β1 had no effect on neointimal formation .CONCLUSION:The AS TGF-β1 specifically inhibits the pro-tein expression of TGF-β1 in the VSMCs derived from injured arteries .Moreover , it significantly inhibits DNA synthesis and cell proliferation, and decreases the expression of FN .Therefore, AS TGF-β1 dramatically attenuates neointima formation after balloon njury .The effects of AS TGF-β1 on the injured VSMCs may be associated with its reverse effects on the altera-tion of VSMC phenotype after balloon injury .

Objective To observe the frequency of interleukin-6(IL-6) gene -572C/G polymorphism and its relationship with myocardial infarction(MI) and serum lipids in Chinese Hans. Methods IL-6/gene-572C/G polymorphism was genotyped in 232 MI patients and 260 healthy adults by PCR-RFLP method. Results There were IL-6/gene-572 CC、CG and GG genotype. -572GG genotype and G allele were more frequent in patients than those in controls(P

BACKGROUND: Hypertension is one of the most important risk factors for stroke, and brain focal renin-angiotensin system has been proved to play a vital role in the development of hypertension and stroke.OBJECTIVE: To observe the influence of long-term administration of losartan, an angiotensin-1 receptor antagonist, on the incidence of strokeprone spontaneous hypertension in rats.DESIGN: Randomized controlled experiment.SETTING: Renmin Hospital Affiliated to Wuhan University.MATERIALS: This experiment was carried out in Wuhan University between July 1999 and March 2001. Totally 26 six-week-old male rats with stroke-prone spontaneous hypertension and 8 Kyoto male Wistar rats were recruited in this experiment with the body mass of 144.5-182.1 g.METHODS: Totally 26 six-week-old male rats with spontaneous hypertension were randomized into stroke-Rrone spontaneous hypertension group (n=9) which received gastric perfusion of physiological saline at a dosage of 5 mL/d; losartan 10 mg/(kg·d) group of 9 rats which received gastric perfusion of losartan at a dosage of 10 mg/(kg ·d) and losartan 30 mg/(kg ·d)group of 8 rats which received gastric perfusion of losartan at a dosage of 30 mg/(kg·d). Rats in the three groups were provided with high-protein feed when entering the group, and drank 15 g/L salty water (5 mL/d) from the onset of week 2. At the same time, 8 six-week-old male Wistar rats were taken as normal controls to receive gastric perfusion of physiological saline at a dosage of 5 mL/d once a day; they took ordinary feed and drank running water. All rats lived with 12 hours' day-night alternation at room temperature of 18-20 ℃ and with humidity of 40%-50%. Totally 18weeks later, the incidence of stroke and BP changes were observed. The clinical manifestation of stroke was scored 1 if rats appeared few activities,with movements slightly reduced or excited; 2 score referred to very few activities, with movements obviously reduced or violently stimulated; 3score referred to inability to walk, lying motionless with melancholy symptoms; score 4 referred to paralysis and inability to stand, lateral or bilateral limb paralysis. Transmission electron microscope was used for histological observation of cell apoptosis in the brain.MAIN OUTCOME MEASURES: ① Observation of brain structure at week 18 when rats were decapitated. ② Results of nerve cell apoptosis detected with TUNEL technique. ③ Rat body mass, BP, as well as the incidence and changes of stroke were recorded.RESULTS: Totally 34 rats entered the result analysis. ① The incidence of stroke in the three groups: It was 100%, 22%, and 13%, respectively, in stroke-prone spontaneous hypertension group, losartan 10 mg/(kg·d) group,and losartan 30 mg/(kg·d) group. ② Score for stroke: The score was remarkably higher in stroke-prone spontaneous hypertension group than in losartan 10 mg/(kg·d) group and losartan 30 mg/(kg·d) group [(3.50±0.55,0.67±1.12, 0.38±0.74) minutes]. ③ Electron-microscopic observation: In stroke-prone spontaneous hypertension group, electron density was found increased in necrotic neurons; moreover, some nuclear membrane lost double-layer structure with ridges broken, even reduced or disappeared, displaying vacuolated changes. In losartan 30 mg/(kg·d) group and losartan 10 mg/(kg·d) group, most of neurons displayed basically normal morphology, with neuron chromatin evenly distributed and nuclear envelops regular, but there were still some neurons that had dense chromatin, with ridges broken and reduced. ④ Nerve cell apoptosis in the three groups: It was found obviously lower in normal group than in losartan 30 mg/(kg ·d)group, losartan 10 mg/(kg·d) group, and stroke-prone spontaneous hypertension group [(2.5±0.8, 13.9±4.3, 14.0±4.4, 52.0±16.7)%, P ＜ 0.05]. ⑤ BP changes: At week 18, BP was obviously lower in normal group than in losartan 30 mg/(kg·d) group, losartan 10 mg/(kg·d) group and strokeprone spontaneous hypertension group [(120.1±7.9, 169.4±10.1,216.7±8.3,225.5±6.8) mmHg (1 mm Hg=0.133 kPa), P ＜ 0.05]. ⑥ Changes of body mass: At week 18, body mass was obviously higher in normal group than in losartan 30 mg/(kg·d) group, losartan 10 mg/(kg·d) group, and stroke-prone spontaneous hypertension group [(313.3 ±10.1, 270.8 ± 10.4,258.7±12.7, 231.0±6.5) g, P ＜ 0.05].CONCLUSION: Losartan can obviously reduce the incidence of stroke and nerve cell apoptosis in rats with spontaneous hypertension, suggesting that losartan as an angiotensin-1 receptor antagonist may prevent and delay the onset of stroke through antagonizing angiotensin I receptor, thus exerting brain-protecting function.

OBJECTIVETo investigate the effects of BmkTXK(beta), a newly purified 'long chain' peptide inhibitor of K(+) channels from the Chinese scorpion Buthus martensi Karsch (BmK), on the electrophysiological properties of isolated rabbit atrial myocytes.

METHODSThe standard whole-cell patch-clamp technique was used to study the effects of multiple concentrations of BmkTXK(beta) on potassium currents and action potentials.

RESULTSBmkTXK(beta) produced concentration-dependent prolongation of action potential duration at 20%, 50%, and 90% repolarization (APD(20,50,90)) without any use-dependence. Meanwhile, it had no significant effect on RMP, APA, or V(max) (n = 9). At a dose of 1 micro mol/L, BmkTXK(beta) decreased I(to) by 41.4% (n = 10, P < 0.01) at a membrane potential of +50 mV [from (13.63 +/- 0.87) pA/pF to (7.98 +/- 0.78) pA/pF]. I(to) was reduced significantly with an IC(50) value of 1.82 micromol/L (95% confidence interval: 1.47 - 2.17 micro mol/L), in a clear concentration-dependent manner. BmkTXK(beta) blocked I(Ks) and I(Ks),tail with an IC(50) of 20.15 micromol/L and a 95% confidence interval of 16.93 - 23.37 micromol/L. At a concentration of 10 micromol/L, BmkTXK(beta) blocked both I(Ks) (mean reduction 37.3% +/- 4.2%, P < 0.01, n = 7) and I(Ks), tail (mean reduction 35.8% +/- 4.1%, P < 0.01, n = 7). At 0 mV, 10 micromol/L BmkTXK(beta) inhibited both I(Kr) (mean reduction 40.5% +/- 2.6%, P < 0.01, n = 6) and I(Kr), tail (mean reduction 42.3% +/- 2.9%, P < 0.01, n = 6). Blocking of I(Kr) by BmkTXK(beta) occurred in a concentration-dependent manner, with an IC(50) of 17.21 micromol/L (95% confidence interval: 14.76-19.66 micromol/L). An absence of effects on I(K1) was observed for BmkTXK(beta), with no change in reversal-potential (n = 6, P > 0.05).

CONCLUSIONSBmkTXK(beta) exerts direct blocking effects on several potassium channels involved in cardiac repolarization, and has a strong effect on prolonging the repolarization of rabbit cardiomyocytes without reverse frequency dependence. This finding suggests that BmkTXK(beta) could be a promising class III drug for anti-arrhythmic therapy without the risk of proarrhythmia.

Action Potentials ; drug effects ; Animals ; Cells, Cultured ; Heart Atria ; Myocardium ; cytology ; Patch-Clamp Techniques ; Rabbits ; Scorpion Venoms ; pharmacology

Objective To explore the relationship of interleukin-6 (IL-6) gene polymorphisms and its serum level with essential hypertension (EH) in Chinese Hans population and investigate the effect of IL-6 gene polymorphisms on serum IL-6 level. Methods The polymorphisms of IL-6 gene, -597 G/A, - 572 C/G and 174 G/C, were checked in 191 EH patients and 197 control subjects by PCR-RELP. Meanwhile their serum IL-6 level was determined by ELISA. Results Only -597 GG genotype and -174 GG genotype of IL-6 gene was found in the test population. There was significant difference in frequencies of - 572 genotype and allele between 2 groups. -572 CG+GG genotype and G allele were more frequent in patients than in controls (P

Objective: Recent studies have shown that cell implantation can replace infarcted myocardium to improve cardiac performance. The present study was designed to investigate the effects of autologous bone marrow mononuclear cell (BM-MNC) implantation into myocardium bordering the infarction with or without induced by 5-azacytidine on cardiac function after acute myocardial infarction (AMI) in rabbits. Methods: AMI was replicated by ligating the left anterior descending coronary artery. Rabbits were randomly divided into three groups: (1) BM -MNC induced by 5-azacytidine implantation (n=7), (2) BM-MNC implantation alo ne (n=7), and (3) AMI control (n=7). In addition, sham-operated (n=5) rabbit s were randomly selected to serve as non-infarction control. Animals for cell im p lantation were received intramyocardial injection of autologous BM-MNC in myoca rdium bordering the infarction, and echocardiography and hemodynamic studies wer e performed to evaluate cardiac function following 28 days of implantation. Results: Compared with the sham-operated group, the left ventricle (LV) end diastolic pressure (LVEDP) was significantly increased (P0.05). Conclusion: BM-MNC induced by 5-azacytidine implantation into myocardium bordering the infarction can significantly improve impaired cardiac function associated with LV remodeling after AMI, however such improvement is not further promoted compared with that in BM-MNC implantation group alone.

Objective To investigate the value of treadmill exercise test and dipyridamole electrocardiography stress test (DP T) in diagnosis of coronary heart diseases (CHD), with an attemp of developing a safe, effective and simple diagnostic method for patients intolerable to the exercise. Methods Fifty two male patients aged from 41 to 75 years, averaged 58.44?8.79 years and with suspected or confirmed CHD were recruited to perform the treadmill exercise test, DP T and coronary angiography, respectively, within one month. Results The sensitivity and specificity of the treadmill exercise test for diagnosing CHD were 100% and 78.95%, respectively, while those of DP T for diagnosing CHD were 79.31% and 65.22%, respectively.As corresponded to the number of sites of coronary artery lesions,the numbers of cases with positive results revealed were 0, 2, 10 and 17 by the treadmill exercise tests and 0, 6, 11 and 15 by DP T, respectively.Based on the criteria of DP T, the sensitivity and specificity symptom, the changes in electrocardiography (ECG) and the combination of the two, for diagnosing CHD were 72.41%, 65.22%; 65.52%, 82.61%;and 79.31%, 65.22%, respectively. Conclusion The sensitivity and specificity of the treadmill exercise test and the DP T in the diagnosis of CHD were similar. More positive cases were revealed in above mentioned tests as the sites and degree of coronary artery lesion increased. DP T was particularly suitable for helping diagnose CHD in patients intolerable to exercise.

OBJECTIVETo investigate the feasibility and features of 3 catheter-mediated approaches of gene transfer into heart, including direct myocardial injection (DMI), coronary artery perfusion (CAP), and intrapericardial cavity injection (ICI).

METHODSFifteen dogs were used, and 0.3 ml (1 x 10(9) pfu) of an adenovirus (Adex1SR LacZ) was injected into the heart by 3 methods. The dogs were killed 5 days following injection, and gene expressions in heart and liver were evaluated by histochemical analysis.

RESULTSThe results showed that (1) the CAP method was relatively less damaging and induced sparse LacZ expression in the myocardium, and the gene expression was also found in both vessels within the myocardium and liver; (2) gene transfer by DMI resulted in intense LacZ expression around the injection accompanied by a local inflammatory response; (3) LacZ expression elicited by ICI was detected in either the inner surface of the parietal pericardium or epicardial surface of the heart, and also in the myocardium underlying the visceral pericardium.

CONCLUSIONThree catheter-mediated methods of gene transfer into the heart may be used and a reasonable approach should be chosen according to purpose.

Adenoviridae ; genetics ; Animals ; Cardiac Catheterization ; Dogs ; Female ; Gene Expression ; Gene Transfer Techniques ; Genetic Vectors ; administration & dosage ; genetics ; Inflammation ; etiology ; Injections ; adverse effects ; methods ; Lac Operon ; genetics ; Liver ; metabolism ; Male ; Myocardium ; metabolism

AIM: To investigate the effect of L-arginine (L-Arg) on intimal proliferation and expression of related cell cycle regulatory factors after vascular injury in rats. METHODS: Rats were divided into three groups :sham operation group, balloon injury group(this group included balloon 48 h,7 d and 14 d subgroup) and balloon+L-Arg group. Neointima area were calculated morphologiocally. The expression of cyclin dependent kinase-2(CDK2),cyclin E and proliferation cell nuclear antigen (PCNA) were measured by means of immunohistochemical technique and computer image analyzer. RESULTS: After vascular balloon injury, the level of plasma NO decreased, CDK2、cyclin E and PCNA expressed in the media at 48 h and in the neointima at 7 d and 14 d but with low and undetected expression in the media, the expression of CDK2, cyclin E and PCNA increased with the intima thickening. Compared with balloon 14 d group, the plasma NO level increased (P＜0.01）, the neointima area reduced by 59.1%(P＜0.01) and the positive expression indexes of CDK2, cyclin E and PCNA decreased by 36.1%, 46.3% and 76.2% respectively in balloon+L-Arg group (P all＜0.01）. CONCLUSION: L-Arg can effectively repress intima proliferation after vascular injury, which may be associated with its inhibiting the proliferation of vascular smooth muscle cell through downregulating the excessive expression of CDK2, cyclin E and PCNA.

Objective Objective To investigate the effects of losartan on neointimal proliferation and macrophage count in neointima.Methods The rabbits underwent abdominal aorta balloon de-endothelialization,then received a 1 5 % cholesterol diet for 8 weeks.Abdominal aorta atherosclerosis model was performed in 16 rabbits.Immediately after operation, losartan was orally administered to the rabbits of the losartan group (10mg?kg -1 ?day -1 ), while the rabbits of the control group were given normal saline. Four weeks later the animals were killed and the excised artery segments were prepared for histomorphological observation and macrophages were investigated by immunohistochemistry analysis.Results Compared with the control group, the IT/MT ratio and IA/MA ratio of the losartan group were significantly reduced(P