[Efficacy and mechanism of local delivery of rapamycin and rapamycin-loaded poly(lactic-co-glycolic) acid nanoparticles on coronary restenosis of injury-stenosis model of minipigs].

OBJECTIVE: To determine whether intramural administration of rapamycin (RPM)-loaded polylactic-polyglycolic acid (PLGA) nanoparticles (NPs) can reduce intimal thickening and affect the mRNA expressions of matrix metalloproteinase (MMP)-2, tissue inhibitor of metalloproteinase (TIMP)-2 and p27(kipl) in a coronary injury-stenosis model of minipigs. METHODS: Twenty eight minipigs were randomly separated into four groups: saline group (n=7), blank PLGA NPs group (5.0 mg/ml)(n=7), RPM group (1.0 mg/ml)(n=7), and RPM-PLGA NPs(5.0 mg/ml)group (n=7), respectively. Different treatments were intracoronary locally delivered via a Dispatch™ catheter for 10 minutes. Serial angiography was performed pre-and post-modeling 30 days and the percent stenosis degree was assessed. Hematoxylin-Eosin (HE) staining, Weigert's resorcin fuchsin staining and picric acid-sirius red staining were used for morphometric analysis. Immunohistochemistry was performed to assess the levels of proliferating cell nuclear antigen (PCNA), MMP-2, and TIMP-2 at early and late time points, respectively. The expression of p27(kip1) mRNA was detected by in situ hybridization staining. RESULTS: Data from 21 minipigs had been collected at the end of the experiment with 6, 4, 5, and 6 from the former mentioned 4 groups, respectively. For the instant injury index, there was no significant difference among the four groups. The percent stenosis degree of RPM-PLGA NPs group was significantly lower than that of the other three groups respectively (all P< 0.05). The neointima area, net external elastic lamina area to external elastic laminal area ratio, and proliferative index of RPM-PLGA NPs group were significantly less than those of the other three groups, with all the P values less than 0.05. The mean value of integral optical density of p27(kip1)mRNA expression of RPM-PLGA group was 0.35 ± 0.06, higher than that of blank PLGA NPs group (0.12 ± 0.05, P< 0.01), saline group (0.16 ± 0.03, P< 0.05), and RPM group (0.15 ± 0.03, P< 0.05), respectively. The MMP-2/TIMP-2 ratio and the positive expression index of PCNA in RPM-PLGA group were lower than that of the other groups (P< 0.05). CONCLUSIONS: Locally delivered rapamycin-loaded PLGA NPs significantly reduces MMP-2/TIMP-2 ratio and PCNA expression, increases p27(kip1) mRNA expression and significantly relieves percent stenosis degree and shows excellent acute procedural results in the minipig interventional coronary artery oversized balloon injury model. The results from minipig model further support that this approach could be a potential clinical procedure for vascular proliferative disease.

Blood pressure reducing effects of piromelatine and melatonin in spontaneously hypertensive rats.

BACKGROUND: Recently, widespread interest has grown regarding melatonin treatment of hypertension including its cardioprotective effects. Studies in rodents indicate that melatonin plays a role in the pathogenesis of hypertension in rats with metabolic syndrome. Piromelatine, a melatonin agonist, serotonin 5-HT-1A and 5-HT-1D agonist and serotonin 5-HT2B antagonist is a multimodal agent with sleep promoting, anti-diabetic, analgesic, anti-neurodegenerative, anxiolytic and antidepressant potential, currently in development for the treatment of insomnia. AIM: In this report we assessed the effects of piromelatine and melatonin treatment on blood pressure in spontaneously hypertensive rats (SHR) and normotensive Wistar-Kyoto (WKY) rats. MATERIALS AND METHODS: Five groups of 12-wk-old rats (10/group) were treated for 5 weeks with a vehicle, piromelatine (5, 15 and 50 mg/kg BW) and melatonin (10 mg/kg BW) and an age-matched WKY control group. Systolic blood pressure (tail-cuff method) was measured weekly at 9:00 a.m. and at 9:00 p.m. The rats body weight, plasma glucose, insulin, triglyceride, adiponectin, total cholesterol, HDL and LDL/VLDL cholesterol were also measured. RESULTS: Our results showed that both piromelatine and melatonin reduced SHR blood pressure significantly both during the morning and the evening. Piromelatine, but not melatonin, also reduced SHR body weight gain and both significantly decreased plasma glucose and insulin levels and increased adiponectin levels. CONCLUSIONS: Piromelatine, similar to melatonin, has an antihypertensive effect and also attenuates body weight, improves metabolic profiles and might be useful in the treatment of hypertension and the metabolic syndrome.

Associated factors for HIV and syphilis infection among men who have sex with men only and men who have sex with both men and women in cities of China.

The aims of this paper were to find out the status of HIV and syphilis infection and to examine the sexual behaviours between men who have sex with men only (MSM/M) and men who have sex with both men and women (MSM/W), as well as to determine the correlates for HIV and syphilis infection among MSM/M and MSM/W, respectively. Among 1693 MSM who participated in the study, the proportions of MSM/M and MSM/W were 82.1% and 17.9%, respectively. The prevalences of HIV infection were 7.0% in MSM/M and 6.6% in MSM/W and the prevalences of syphilis infection were 11.9% and 13.2%, respectively. Among the MSM/M subset, the correlates both for HIV and syphilis infection included having more sexual partners, and being receptive or both insertive and receptive for anal sex. Among the MSM/W subset, living in Chengdu was associated with HIV infection and using condoms inconsistently during anal sex was associated with syphilis infection. The findings of this survey call for interventions tailored according to the needs of different subsets of MSM.

Enhancement of manumycin A-induced apoptosis by methoxyamine in myeloid leukemia cells.

Farnesyltransferase inhibitors (FTIs) are currently under investigation for leukemia treatment. We evaluated the FTI manumycin A (manumycin) in two myeloid leukemia cell lines (U937 and HL-60). Manumycin induced nitric oxide production and apoptosis of the leukemia cells. Nitric oxide or other reactive oxygen species may induce oxidative DNA damage, and the number of apurinic sites increased after manumycin treatment, which was reversed by concurrent treatment with N-acetyl-L-cysteine. Since repair of DNA damage is important to cell survival, we hypothesized that methoxyamine, an inhibitor of base-excision repair, would enhance the antineoplastic effect of manumycin. The combination of manumycin and methoxyamine resulted in enhanced apoptosis by six criteria increased annexin V binding, release of mitochondrial cytochrome c into the cytosol, activation of caspase-9, activation of caspase-3, specific cleavage of poly-adenosyl ribose polymerase, and increase in the sub-G1 cell cycle fraction. The drug combination enhanced inhibition on the soft agar clonogenic assay and on the formazan dye cell viability assay. The effects of manumycin or manumycin plus methoxyamine on apoptosis were blocked by N-acetyl-L-cysteine, and partially by nitric oxide synthase inhibitors or scavenger of peroxide. We conclude that methoxyamine enhances manumycin-induced apoptosis in myeloid leukemia cells.

Association between carcinogen-DNA adducts in white blood cells and lung cancer risk in the physicians health study.

In this matched case-control study nested within the prospective Physicians' Health Study, we evaluated whether DNA damage in blood samples collected at enrollment significantly predicted risk, consistent with our hypothesis that cases have greater biological susceptibility to polycyclic aromatic hydrocarbons and other aromatic tobacco carcinogens. The subjects were 89 cases of primary lung cancer and 173 controls, all males, matched on smoking, age, and duration of follow-up. Aromatic-DNA adducts were measured in WBCs by the nuclease P1-enhanced (32)P-postlabeling method that primarily detects smoking-related adducts. Among current smokers, but not former or nonsmokers, there was a significant increase in mean adduct levels of cases compared with controls (11.04 versus 5.63; P = 0.03). "Healthy" current smokers who had elevated levels of aromatic DNA adducts in WBCs were approximately three times more likely to be diagnosed with lung cancer 1-13 years later than current smokers with lower adduct concentrations (odds ratio, 2.98; 95% confidence interval, 1.05-8.42; P = 0.04). We were not able to discern case-control differences in former smokers and nonsmokers. The findings are of interest because they suggest that individuals who become cases have greater biological susceptibility to tobacco carcinogens, a biological difference, which manifests most clearly while exposure is ongoing.

Development of serine protease inhibitors displaying a multicentered short (<2.3 A) hydrogen bond binding mode: inhibitors of urokinase-type plasminogen activator and factor Xa.

Novel scaffolds that bind to serine proteases through a unique network of short hydrogen bonds to the catalytic Ser195 have been developed. The resulting potent serine protease inhibitors were designed from lead molecule 2-(2-hydroxyphenyl)1H-benzoimidazole-5-carboxamidine, 6b, which is known to display several modes of binding. For instance, 6b can recruit zinc and bind in a manner similar to that reported by bis(5-amidino-2-benzimidazolyl)methane (BABIM) (Nature 1998, 391, 608-612).(1) Alternatively, 6b can bind in the absence of zinc through a multicentered network of short (<2.3 A) hydrogen bonds. The lead structure was optimized in the zinc-independent binding mode toward a panel of six human serine proteases to yield optimized inhibitors such as 2-(3-bromo-2-hydroxy-5-methylphenyl)-1H-indole-5-carboxamidine, 22a, and 2-(2-hydroxybiphenyl-3-yl)-1H-indole-5-carboxamidine, 22f. Structure-activity relationships determined that, apart from the amidine function, an indole or benzimidazole and an ortho substituted phenol group were also essential components for optimal potency. The affinities (K(i)) of 22a and 22f, for example, bearing these groups ranged from 8 to 600 nM toward a panel of six human serine proteases. High-resolution crystal structures revealed that the binding mode of these molecules in several of the enzymes was identical to that of 6b and involved short (<2.3 A) hydrogen bonds among the inhibitor hydroxyl oxygen, Ser195, and a water molecule trapped in the oxyanion hole. In summation, novel and potent trypsin-like serine protease inhibitors possessing a unique mode of binding have been discovered.

Addressing the stability of C-capped dipeptide efflux pump inhibitors that potentiate the activity of levofloxacin in Pseudomonas aeruginosa.

Synthetic optimization of a biologically labile class of dipeptides that function as efflux pump inhibitors to potentiate the antibacterial agent levofloxacin in Pseudomonas aeruginosa has led to the discovery of a related series of compounds that are completely stable in a variety of biological matrices. Other than the stability profile, the in vitro profile of the new series is essentially identical to that observed with the original one. A prototypical compound from the new series demonstrates potentiation in an in vivo model of infection.

[Effect of mm-LDL on NF-kB activation in endothelial cell].

OBJECTIVE: To investigate the signal transduction pathway of NF-kB activated by minimally modified low density lipoprotein (mm-LDL) in endothelial cells and the effect of NF-kB on platelet derived growth factor b (PDGFb) mRNA expression. METHODS: mm-LDL was prepared through iron oxidation by dialyzing the native LDL against FeSO4 in PBS. Endothelial cells were incubated in a medium containing mm-LDL, TNF, and IL-1 respectively and electrophoretic mobility shift assay (EMSA) was displayed to check on the activation of NF-kB. Luciferase reporter gene was analysed to investigate the effect of nuclear factor inducing kinase (NIK), inhibitor of NF-kB kinase alpha (IKK alpha) and inhibitor of NF-kB kinase beta (IKK beta) on NF-kB activation. In addition, endothelial cells were transfected using PDGFb promoter-luciferase for reporter gene analysis or transfected with mut-NIK for slot blot analysis to study the effect of NF-kB on PDGFb mRNA expression. RESULTS: mm-LDL was able to activate NF-kB in endothelial cells. mut-NIK and mut-IKK beta inhibited luciferase activity induced by mm-LDL. mm-LDL could also enhance luciferase activity controlled by upstream sequence of PDGFb promoter which contains element interacting with NF-kB. Result of slot blot showed inhibition of PDGFb mRNA expression by mut-NIK in the endothelial cells stimulated by mm-LDL. CONCLUSION: mm-LDL may activate NF-kB through NIK-IKK beta pathway and promote PDGFb mRNA expression in endothelial cells.

[Effect of oxLDL on the uptake and clearance rate of cholesterol in vascular smooth muscle cells originated from human apoAI transgenic mice].

OBJECTIVE: Study on (1) inhibition of oxidized low density lipoprotein (oxLDL) effect on the uptake and clearance of intra-cellular 3H-cholesterol in vascular smooth muscle cells (v-SMC) originated from the human-apoAI transgenic mice (C57BL/6); (2) change of human-apolipoprotein AI (h-apoAI) mRNA expression in v-SMC after oxLDL stimulation and the protective effect of expressed h-apoAI on v-SMC against oxLDL intoxication. METHODS: (1) v-SMC isolated from human apoAI transgenic mice possessing a recombined gene connected beforehand with a mouse metallothionein-I (MT-I) as the promoter; (2) study of h-apoAI mRNA expression from v-SMC of the transgenic mice by RT-PCR and Northern blot. RESULTS: oxLDL (30 micrograms/ml) strongly promoted v-SMC proliferation. No difference found on 3H-cholesterol uptake between smooth muscle from normal mouse aorta (n-SMC) and smooth muscle cells from transgenic mouse aorta (tr-SMC) of the control groups, the uptake rates of both kinds of SMC rose 100% after oxLDL stimulation. The efflux rates of 3H-cholesterol in tr-SMC were much higher than those of n-SMC (40%-50%). After oxLDL stimulation, the clearance rates fell by 28% and 10% respectively for n-SMC and tr-SMC. The result of RT-PCR and Northern blot showed a marked increase of h-apoAI gene expression after oxLDL stimulation. CONCLUSIONS: Expression of h-apoAI gene in C57BL/6 mice enables to decrease the accumulation of cholesterol in v-SMC. tr-SMC are capable to alleviate the harmful effect of oxLDL on v-SMC due to the increase of h-apoAI expression.

[Activation of NF-kappa B and its effect on induction of platelet-derived growth factor-B transcription by angiotensin II in ECV304 cell line].

OBJECTIVES: The renin-angiotensin system may contribute to the pathogenesis of atherosclerosis. The transcription factor nuclear factor-kappa B (NF-kappa B) participates in most signal pathways involved in the inflammatory process. In this project the effect of angiotensin II (Ang II) on NF-kappa B activation and the promotion of PDGF-B mRNA expression in human endothelial cell line ECV304 was studied. METHODS: Electrophoretic mobility shift assay, immunofluorescence and immunoelectronic microscope techniques, including confocal microscopy and gold particle labelled electronic microscopy were applied to investigate the mechanism by which Ang II activates NF-kappa B, ECV304 cells were transiently transfected with an NF-kappa B/luciferase reporter gene and catalytically inactive NIK, IKK alpha, IKK beta mutants respectively. Northern blot was carried out to detect PDGF-B mRNA. RESULTS: By the findings of immunofluorescence confocal microscopy, immunoelectronic microscopy and Northern blot, Ang II was effective in stimulating NF-kappa B activation and there was definited cytoplasmic-to-nuclear translocation of NF-kappa B subunits p50 and p65 and overexpression of PDGF-B mRNA expression. Over-expression of the transiently transfected IKK alpha-KM, IKK beta-KM and NIK-KM mutant genes enabled to block the reporter gene activation induced by ang II. CONCLUSION: Ang II is effective to activate NF-kappa B through a pathway dependent on NIK, IKK alpha and IKK beta, and induces PDGF-B transcription in the endothelial cell line ECV304.

Molecular basis for the effect of lipid lowering drugs on growth factors after de-endothelialization.

OBJECTIVE: To study the mechanism and effect of lipid lowering drugs in arresting the development of arterial restenosis after angioplasty. METHODS: De-endothelialization injury of rabbit aortae, common iliac and femoral arteries using balloon angioplasty and the expression of growth factors such as platelet derived growth factor-B (PDGF-B), transforming growth factor beta-1 (TGF beta-1), and fibroblast growth factors (bFGF) were investigated. Total serum cholesterol (TC) and triglycerides (TG) were analyzed during and after the treatment using either simvastatin combined with gemfibrozil or simvastatin alone for 6 weeks. RESULTS: Serum total cholesterol and triglycerides were only slightly to moderately increased after high cholesterol ration intake lasting for 6 weeks in rabbits of two therapeutic groups (simvastatin plus gemfibrozil or only simvastatin). A positive correlation was found between TC and intimal/medial ratio (r = 0.5873, P < 0.05). PDGF-B detected by immuno-histochemistry and RT-PCR analysis showed that the release of PDGF-B was inhibited by simvastatin and gemfibrozil after de-endothelialization. RT-PCR analysis showed that TGF beta-1 was increased in the neointima in two treatment groups but no definite change was seen in the mRNA of bFGF in the smooth muscle cell (SMC) of the balloon-injured arteries even under lipid lowering drug treatment. CONCLUSION: In addition to the lipid lowering effect, both simvastatin and gemfibrozil also influence the release of PDGF-Band TGF-1 in the neointima after de-endothelialization.

Suppression of induced atherosclerosis in h-apo AI transgenic mice by overexpression of human apo AI in the aortic wall.

OBJECTIVES: To investigate the inhibitory effect of expressed human apolipoprotein AI (h-apo AI) and high density lipoprotein (HDL) on atherosclerosis development in transgenic mice, and cultivation of smooth muscle cells isolated from the aortic wall of transgenic mice that are able to produce human apo AI in vitro. METHODS: Both h-apo AI transgenic mice and normal C57 mice were fed with either a regular chow or a high-fat diet containing 5% pork lard, 1.25% cholesterol and 0.25% sodium cholate for 14 or 24 weeks respectively. Human apo AI mRNA were detected by Northern blot. Plasma apo AI levels were measured using a radio-immuno-diffusion assay, and plasma lipid levels were measured using a colorimetric assay. Image analysis was performed in order to quantify the fatty streak areas stained with oil red O. In addition, smooth muscle cells isolated from the media layer of the aortic wall of h-apo AI transgenic mice were cultured for the detection of human apo AI produced. RESULTS: Higher levels of h-apo AI mRNA were found in liver, small intestine, kidneys and aortae in transgenic mice than in the controls all on a high-fat diet. The transgenic mice had an increased level of serum apo AI and HDL-cholesterol and the fatty streak area counted at the aortic sinus was approximately 5-fold less in the transgenic mice after feeding with a high fat ration, particularly after 24 weeks. SMC isolated from the transgenic mice aortae were cultivated and able to express h-apo AI mRNA and its related protein. CONCLUSION: Elevation of h-apo AI and HDL in serum and aortic wall of the transgenic mice has a remarkably inhibitory effect on the development of experimental atherosclerosis.

Interaction of alcohol and the G to a substitution at the promoter region of the apolipoprotein AI gene in determining plasma apolipoprotein AI levels in Yi and Han Chinese.

OBJECTIVE: To investigate the influence on plasma lipid levels of alcohol and a common polymorphism in the human apolipoprotein AI gene promoter at a position 75 bp upstream of the transcription start site. METHODS: For this study, 742 healthy Yi and Han subjects all above 15 years old formed the total population which was divided into three groups: the Yi-farmer group, the Yi-emigrant group and the Han-resident group. All estimates of plasma lipids and apolipoproteins were performed using an auto-analyzer. Genetic DNA was prepared from the blood clots using the Triton X-100 lysis technique. Amplification of a 432 bp fragment of the apoAI gene promoter was performed using PCR followed by restriction digestion, electrophoresis and identification of the genotypes involved. RESULTS: The samples were divided on the basis of alcohol consumption: non-drinkers, 1-25 g/day, 26-75 g/day and > 75 g/day. Comparing the four alcohol consumption groups, plasma HDLC and apoAI levels were increased as the alcohol consumption increased with no evidence of threshold effects in the Yi-farmers and the Han people groups. A similar association was found in the Yi-emigrant group, but was not statistically significant. The frequencies of the A allele in the three populations were similar, and no significant difference of lipid and apolipoprotein levels was found between subjects with and without the A allele in the three populations. But, in Han and Yi-emigrant samples, the drinkers with the GG genotype had higher plasma HDLC and apoAI levels than non-drinkers with the same genotype, while the drinkers with the A allele had lower plasma HDLC and apoAI levels than drinkers without the A allele. Non-drinkers with the A allele had higher levels of apoAI than non-drinkers with GG genotypes. It was estimated that 18% of the variability of plasma apoAI level could be explained by the G to A polymorphism in non-drinkers of Yi-emigrants (F = 8.94, P < 0.01). CONCLUSIONS: The present data suggest that moderate alcohol consumption and the G to A substitution could lower the risk of coronary heart disease (CHD), but the beneficial effects of one will be negated by the other.

[Inhibition of monocytes adhesion to the intima of arterial wall by local expression of antisense monocyte chemotactic protein-1].

OBJECTIVE: To study the mechanism of monocyte recruitment in atherogenesis and to clarify the effect of monocyte chemotactic protein-1 (MCP-1) in this process. METHODS: Femoral arteries isolated from the rabbits which had been fed with a high cholesterol diet and locally perfused with MM-LDL within the artery beforehand, were used as the models. Antisense MCP-1cDNA was transferred into the arterial wall by injecting recombinant LNCX-anti-MCP-1/liposomal complex in the femoral sheath and the periarterial tissue. RESULTS: Expression of antisense MCP-1 mediated by recombinant LNCX plasmid/lipsomal complex gene transfer enabled to inhibit MCP-1 gene expression and adhesion of monocyte to the intima. CONCLUSION: MCP-1 plays an important role on the recruitment of monocytes in the arterial wall, which provides a potential clue in developing a gene therapy project for the prevention and treatment of atherogenesis.

[Influence of a G/A substitution at -75 bp promoter region of apolipoprotein A I gene and wine consumption on the plasma apolipoprotein A I levels in Yi and Han Chinese].

OBJECTIVE: To study the influence of a common polymorphism of the human apolipoprotein A I gene promoter at a position of -75 bp upstream of the transcription start site and wine consumption on plasma lipid levels. METHODS: 742 healthy Yi and Han healthy subjects all above 15 years old to be the total population which was divided into three samples, namely, Yi-farmer sample, Yi-emigrant sample and Han-resident sample for this study. Estimations of plasma lipids and apolipoproteins were carried on through an auto-analyzer. Genetic DNA was prepared from the frozen blood clot using Triton x-100 lysis technique. Amplification of a 432 bp fragment of the apoA I gene promoter was performed via the PCR, followed by restriction digestion, electrophoresis and identification of the genotypes involved. Data analysis was done at last. RESULTS: Four groups of alcohol consumption were defined: the non-drinkers, 1 - 25 g alcohol intake/day, 26 - 75 g alcohol/day and > 75 g alcohol/day. A tendency of a mild persistent elevation of plasma HDL-C and apoA I was noticed corresponding to an increase of the amount of alcohol intake, and with no evidence of threshold effect observed in the samples of both the Yi-farmers and the Han people. Similar phenomenon was obtained in the sample of Yi-emigrants, but no statistical significance. The frequencies of the A allele of all 3 samples were similar. In Han and Yi-emigrant samples, the drinkers with genotypes of GG had a higher plasma HDL-C and apoA I level than that of the non-drinkers with the same genotypes. Drinkers with A allele had a lower plasma HDL-C and apoA I level than that of drinkers without A allele, and the non-drinkers with A allele had a higher levels of apoA I than of non-drinkers with genotypes of GG. It is estimated that 18% of the variability of plasma apoA I level could be explained by the G/A polymorphism in non-drinkers of Yi-emigrants (F = 8.94, P < 0.01). CONCLUSIONS: Current data suggest that a moderate alcohol consumption or a G to A substitution could make a lower incidence of CHD, but the beneficial effect of one will be negated by the other when both factors occur simultaneously. This finding is seemed valuable for a further study on the effect of the environmental factor or genetic factor in effecting the plasma apoA I level afterwards.

Factors contributing to troponin exchange in myofibrils and in solution.

The troponin complex in a muscle fiber can be replaced with exogenous troponin by using a gentle exchange procedure in which the actin-tropomyosin complex is never devoid of a full complement of troponin (Brenner et al. (1999) Biophys J 77: 2677-2691). The mechanism of this exchange process and the factors that influence this exchange are poorly understood. In this study, the exchange process has now been examined in myofibrils and in solution. In myofibrils under rigor conditions, troponin exchange occurred preferentially in the region of overlap between actin and myosin when the free Ca2+ concentration was low. At higher concentrations of Ca2+, the exchange occurred uniformly along the actin. Ca2+ also accelerated troponin exchange in solution but the effect of S1 could not be confirmed in solution experiments. The rate of exchange in solution was insensitive to moderate changes in pH or ionic strength. Increasing the temperature resulted in a two-fold increase in rate with each 10 degrees C increase in temperature. A sequential two step model of troponin binding to actin-tropomyosin could simulate the observed association and dissociation transients. In the absence of Ca2+ or rigor S1, the following rate constants could describe the binding process: k1 = 7.12 microM(-1) s(-1), k(-1) = 0.65 s(-1), k2 = 0.07 s(-1), k(-2) = 0.0014 s(-1). The slow rate of detachment of troponin from actin (k(-2)) limits the rate of exchange in solution and most likely contributes to the slow rate of exchange in fibers.

Inhibition of intimal lipid deposition in human apolipoprotein A1 transgenic mice.

1. Southern blot confirmed the emergence of the human apolipoprotein A1 (h-apoA1) gene (four to 15 copies) in newborn transgenic mice. The inheritance pattern of h-apoA1 transgene in three generations of progeny was compatible with a single autosomal integration site. 2. Northern blot showed h-apoA1 mRNA expressed mainly in liver, kidney and aorta. The total plasma apoA1 (murine plus human) level was significantly increased to 58 and 118 in transgenic mice without or under zinc induction, respectively. The majority was h-apoA1. Plasma lecithin:cholesterol acyltransferase activity was positively correlated with h-apoA1 levels in transgenic mice (r = 0.43; P < 0.01). 3. After a high cholesterol intake for 14 weeks, the incidence of fatty streak at the aortic sinus was 40% in transgenic mice and 80% in controls. Expression of platelet-derived growth factor-B, c-myc and c-fos proteins was much weaker in smooth muscle cells at the aortic sinus in transgenic mice.

[The modulation of collagen synthesis and expressions of type I and III collagen mRNAs in aorta smooth muscle cell by PDGF and heparin].

OBJECTIVE: The effect of platelet derived growth factor (PDGF) and heparin on the modulation of human aorta smooth muscle cell(hASMC) proliferation, collagen synthesis, expressions of type I and III collagen mRNAs as well as transforming growth factor-beta (TGF-beta) mRNA were investigated. METHODS: 3H-TdR and 3H-proline incorporation and Northern blot analysis were done. 3H-TdR and 3H-proline incorporation was statistically analyzed with t test among different experimental groups. RESULTS: In comparison to the control group, PDGF possessed the ability in promoting markedly the DNA synthesis (3H-TdR incorporation: PDGF group vs control, P < 0.01), synthesis and secretion of collagen protein (3H-proline incorporation of SMC and medium: PDGF group vs control, P < 0.01), expressions of type I and type III collagen mRNAs, and transforming growth factor mRNA in hASMC. Whereas heparin significantly decreased the DNA synthesis (heparin group vs control, P < 0.01), synthesis and secretion of collagen protein (heparin group vs control, P < 0.01) of hASMC in vitro. Heparin also inhibited the promoting effect of PDGF on DNA synthesis, the synthesis and secretion of collagen protein, and reduced the up-regulation of expressions of type I and type III collagen mRNAs as well as TGF-beta mRNA that stimulated by PDGF. CONCLUSIONS: PDGF could promote the collagen synthesis through upregulating type I and type III collagen mRNA expressions while heparin just inhibiting SMC DNA and collagen synthesis effected in the way of anti-atherosclerosis.

[Morphometric study of inflammatory cells and smooth muscle cells in coronary plaques and their relation with plaque stability].

OBJECTIVE: To compare the content of inflammatory cells and smooth muscle cells in stable and unstable coronary plaques in order to clarify the role of these cells in the vulnerability of plaques. METHODS: Hearts from 12 postmortem acute myocardial infarction cases were studied. The epicardial coronary arteries were dissected en bloc, fixed, decalcified, cut at 4 mm intervals and routinely processed for HE sections. 163 stable plaques (with no or only little lipid core) and 163 unstable plaques (with lipid core size > 40% of plaque area) were studied immunohistochemically using monoclonal antibodies specific for macrophages (CD68), T lymphocytes (UCHL-1) and smooth muscle cells (actin). Computor aided planimetry was used to measure the positive area of different cells. RESULTS: The content of macrophage and T lymphocytes in unstable plaques were significantly higher than that in stable plaques (P < 0.05). CONCLUSIONS: Stable plaques and unstable plaques not only had different lipid core size but also had different inflammatory cell and smooth muscle cell content. The larger the lipid core and the more macrophages and T lymphocytes in the fibrous cap, the more unstable the plaque. The lipid core and inflammatory cells (including macrophages and T lymphocytes) are the two major determinants of the vulnerability of coronary plaques.