Abstract:Aim To investigate cholesterol-induced atherogenesis molecular mechanism, clone and analyse the differentially expressed genes in vascular endothelial cell induced by cholesterol. Methods Suppression subtractive hybridization (SSH) method is used to isolate the differentially expressed cDNA in human umbilical vein endothelial cell induced by cholesterol. After sequencing and homology analysis, ten differentially expressed cDNA fragments are selected and analysed by reverse transcription-polymerase chain reaction(RT-PCR). Results Twenty-three differentially expressed cDNA fragments have been isolated. Seventeen of them are known genes, the other six are unknown genes. The known genes include thrombospondin-1, proteasome subunit (type and so on). The ten elective cDNA fragments (including four known genes and six unknown genes) that are identified by RT-PCR have the same expressed tendency as SSH discovery. Six differentially expressed unknown cDNA fragments have been accepted by GenBank as novel expressed sequence-tags (EST). Conclusion The high level expression of thrombospondin-1 and the low level expression of proteasome subunit (type) in endothelial cell induced by cholesterol may be related to cholesterol-induced atherogenesis.

Abstract:Aim To investigate cholesterol-induced atherogenesis molecular mechanism, clone and analyse the differentially expressed genes in vascular endothelial cell induced by cholesterol. Methods Suppression subtractive hybridization (SSH) method is used to isolate the differentially expressed cDNA in human umbilical vein endothelial cell induced by cholesterol. After sequencing and homology analysis, ten differentially expressed cDNA fragments are selected and analysed by reverse transcription-polymerase chain reaction(RT-PCR). Results Twenty-three differentially expressed cDNA fragments have been isolated. Seventeen of them are known genes, the other six are unknown genes. The known genes include thrombospondin-1, proteasome subunit (type and so on). The ten elective cDNA fragments (including four known genes and six unknown genes) that are identified by RT-PCR have the same expressed tendency as SSH discovery. Six differentially expressed unknown cDNA fragments have been accepted by GenBank as novel expressed sequence-tags (EST). Conclusion The high level expression of thrombospondin-1 and the low level expression of proteasome subunit (type) in endothelial cell induced by cholesterol may be related to cholesterol-induced atherogenesis.

Abstract:Aim Cloning of genes that differentially expressed in remodeling thoracic aorta of 2K1C hypertensive rats(2-kidney, 1-clip Goldblatt rats), which is important for understanding the molecular basis of hypertensive vascular remodeling. Methods We used suppression subtractive hybridization (SSH) to isolate differentially expressed EST in remodeling thoracic aorta. This led to identification of more than fifteen differential expressed sequence-tags(EST). After further cloned and sequenced and homology searched in Genbank with software Stand BLAST. Some differentially displayed genes coding protein were confirmed by western blot. Results We have obtained fifteen pieces of differentially displayed EST, such as cytochrome C and Bcl-2. Moreover, the results from Western Blot confirmed that cytochrome C is highly expressed in in remodeling thoracic aorta and Bcl-2 lowly expressed in remodeling thoracic aorta. Conclusion Cytochrome C is released from mitochondria in response to a variety of apoptotic stimuli, such as reactive oxygen radicals, calcium and ceramide. The Bcl-2 family of proteins serves as critical regulators of pathways involved in apoptosis. Bcl-2 protein is anti-apoptotic. Our data indicate that cell apoptotic mechanism may have important role in hypertensive vascular remodeling.

Abstract:Aim Cloning of genes that differentially expressed in remodeling thoracic aorta of 2K1C hypertensive rats(2-kidney, 1-clip Goldblatt rats), which is important for understanding the molecular basis of hypertensive vascular remodeling. Methods We used suppression subtractive hybridization (SSH) to isolate differentially expressed EST in remodeling thoracic aorta. This led to identification of more than fifteen differential expressed sequence-tags(EST). After further cloned and sequenced and homology searched in Genbank with software Stand BLAST. Some differentially displayed genes coding protein were confirmed by western blot. Results We have obtained fifteen pieces of differentially displayed EST, such as cytochrome C and Bcl-2. Moreover, the results from Western Blot confirmed that cytochrome C is highly expressed in in remodeling thoracic aorta and Bcl-2 lowly expressed in remodeling thoracic aorta. Conclusion Cytochrome C is released from mitochondria in response to a variety of apoptotic stimuli, such as reactive oxygen radicals, calcium and ceramide. The Bcl-2 family of proteins serves as critical regulators of pathways involved in apoptosis. Bcl-2 protein is anti-apoptotic. Our data indicate that cell apoptotic mechanism may have important role in hypertensive vascular remodeling.

Abstract:Aim To investigate whether homocysteine (HCY) induce the expression of secretory macrophage inflammatory protein-1α (MIP-1α)in THP-1 monocytes. Methods After exposure of the cultured THP-1 monocytes to HCY at increasing concentrations (0.05, 0.1 and 0.2 mmol/L) for 8 h, or at a concentration of 0.1 mmol/L for different incubation times (4, 8 and 16 h), enzyme-linked immunosorbent assay (ELISA) was used to determine MIP-1α protein expression in the cultured supernate of each group. Conditioned media with or without HCY (0.01 mmol/L) were collected for chemotaxis assay by micropore filter method using a modified Boyden Chamber. Results ELISA showed that the exposure of THP-1 monocytes to HCY at different concentrations and for different times induced higher production of MIP-1α protein compared with the control (p<0.01). The MIP-1α protein in cultured supernate was increased in a dose- and time-dependent manner. In chemotaxis assay, the migration distance of monocytes in HCY stimulating group was 83.2±4.8 μm, which was significantly longer than that in the non-HCY stimulating group (73.2±2.3 μm) and random migration group (62.4±3.5 μm)(F=310.70, p<0.01). After adding goat anti-human MIP-1α antibody, the monocyte chemotactic activity was noticeably inhibited, compared with the HCY stimulating group without specific antibody (p<0.01). Conclusions HCY can induce THP-1 monocytes to express MIP-1αand secret into culture medium, which share partial chemotactic activity for peripheral monocytes, and may play an important role in atherogenesis through promoting the recruitment of monocytes into arterial intima.

Abstract:Aim To investigate whether homocysteine (HCY) induce the expression of secretory macrophage inflammatory protein-1α (MIP-1α)in THP-1 monocytes. Methods After exposure of the cultured THP-1 monocytes to HCY at increasing concentrations (0.05, 0.1 and 0.2 mmol/L) for 8 h, or at a concentration of 0.1 mmol/L for different incubation times (4, 8 and 16 h), enzyme-linked immunosorbent assay (ELISA) was used to determine MIP-1α protein expression in the cultured supernate of each group. Conditioned media with or without HCY (0.01 mmol/L) were collected for chemotaxis assay by micropore filter method using a modified Boyden Chamber. Results ELISA showed that the exposure of THP-1 monocytes to HCY at different concentrations and for different times induced higher production of MIP-1α protein compared with the control (p<0.01). The MIP-1α protein in cultured supernate was increased in a dose- and time-dependent manner. In chemotaxis assay, the migration distance of monocytes in HCY stimulating group was 83.2±4.8 μm, which was significantly longer than that in the non-HCY stimulating group (73.2±2.3 μm) and random migration group (62.4±3.5 μm)(F=310.70, p<0.01). After adding goat anti-human MIP-1α antibody, the monocyte chemotactic activity was noticeably inhibited, compared with the HCY stimulating group without specific antibody (p<0.01). Conclusions HCY can induce THP-1 monocytes to express MIP-1αand secret into culture medium, which share partial chemotactic activity for peripheral monocytes, and may play an important role in atherogenesis through promoting the recruitment of monocytes into arterial intima.

Abstract:Aim To investigate high quantity of total soy isoflavones influence adhesion molecule such as intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) expression in aorta vessel during the formation of atherosclerosis in rats predisposed with high cholesterol and high fat diet. Methods 60 Wistar rats were divided in 6 groups randomly, rats of the normal diet control (NDC)group were fed with normal diet. Rats of the remainder groups were fed high cholesterol (3.5%), high fat (10%) and 1% hyocholic salt plus 85% normal diet. Rats of the high fat diet plus low (HLI group), medium (HMI group), high (HHI group) dose ISO groups were fed with high content of total isoflavones simultaneously and the dosage were 30 mg/kg, 90 mg/kg and 270 mg/kg body weights perday; rats of the high fat diet plus estrobene control (HEC) group was given estrobene 0.25 mg/kg body weights perday. After 20 week, rats were killed and aorta were taken. HE staining was used to study the pathology during the formation of atherosclerosis. Body weights and organ weight were detected to explore if there were physical impairmen when isoflavones were given to rats. Immunohistochemistry and Western-blotting were used to detect ICAM-1 and VCAM-1 gene expression in aorta vessel and computer image analysis was used to check the degree of gene expression about VCAM-1 or ICAM-1. Results Soy isoflaones can inhibits atherosclerotic plaque formation dose dependently, lower the high expression of ICAM-1 and VCAM-1 in aorta vessel caused by high cholesterol and high fat diet (Compared to B grouPp<0.05). Conclusion Soy isoflavones can inhibit expression of adhesion molecules genes such as ICAM-1, VCAM-1 in aortic of rats fed with high cholesterol and high fat diets.

Abstract:Aim To investigate high quantity of total soy isoflavones influence adhesion molecule such as intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) expression in aorta vessel during the formation of atherosclerosis in rats predisposed with high cholesterol and high fat diet. Methods 60 Wistar rats were divided in 6 groups randomly, rats of the normal diet control (NDC)group were fed with normal diet. Rats of the remainder groups were fed high cholesterol (3.5%), high fat (10%) and 1% hyocholic salt plus 85% normal diet. Rats of the high fat diet plus low (HLI group), medium (HMI group), high (HHI group) dose ISO groups were fed with high content of total isoflavones simultaneously and the dosage were 30 mg/kg, 90 mg/kg and 270 mg/kg body weights perday; rats of the high fat diet plus estrobene control (HEC) group was given estrobene 0.25 mg/kg body weights perday. After 20 week, rats were killed and aorta were taken. HE staining was used to study the pathology during the formation of atherosclerosis. Body weights and organ weight were detected to explore if there were physical impairmen when isoflavones were given to rats. Immunohistochemistry and Western-blotting were used to detect ICAM-1 and VCAM-1 gene expression in aorta vessel and computer image analysis was used to check the degree of gene expression about VCAM-1 or ICAM-1. Results Soy isoflaones can inhibits atherosclerotic plaque formation dose dependently, lower the high expression of ICAM-1 and VCAM-1 in aorta vessel caused by high cholesterol and high fat diet (Compared to B grouPp<0.05). Conclusion Soy isoflavones can inhibit expression of adhesion molecules genes such as ICAM-1, VCAM-1 in aortic of rats fed with high cholesterol and high fat diets.

Abstract:Aim To investigate the effects and mechanisms of losartan, a specific blocker of angiotensin Ⅱ(Ang Ⅱ) receptor-1 (AT 1R) on vascular remodeling of two-kidney, one-cliped hypertensive rat. Methods The blood pressure (BP),heart weight (HW),and body weight (BW) were determined. The morphologic change of aortas and mesenteric resistance arteries was observed after staining with HE. Western blot was performed to valuate the ERK1/2 activity of aortas. Results The blood pressure,and HW/BW in hypertension group were significantly larger than that of control group (178±17 mm Hg vs 119±11 mm Hg, 0.40±0.07 vs 0.27±0.01 respectively, both p<0.01). Treatment with 20 mg/(kg·d) of losartan normalized BP(132±9 mmHg vs 178±17 mmHg,p<0.05) and HW/BW (0.32±0.03 vs 0.40±0.07,p<0.05). The lumen to media of mesenteric resistance arteries was decreased compared with control group (6.00±0.89 vs 8.96±1.23, p<0.01). Losartan normalized the lumen to media of hypertension group (8.87±1.25 vs 6.00±0.89,p<0.05). The p-ERK1/2 of aortas in hypertension group was overexpression compared with control group, and the overexpression of p-ERK1/2 was downregulated by treatment with losartan. Conclusion Losartan could regulate vascular remodeling in 2-kidney,1-cliped hypertensive rat by inhibiting the signaling pathway of ERK1/2.

Abstract:Aim To investigate the effects and mechanisms of losartan, a specific blocker of angiotensin Ⅱ(Ang Ⅱ) receptor-1 (AT 1R) on vascular remodeling of two-kidney, one-cliped hypertensive rat. Methods The blood pressure (BP),heart weight (HW),and body weight (BW) were determined. The morphologic change of aortas and mesenteric resistance arteries was observed after staining with HE. Western blot was performed to valuate the ERK1/2 activity of aortas. Results The blood pressure,and HW/BW in hypertension group were significantly larger than that of control group (178±17 mm Hg vs 119±11 mm Hg, 0.40±0.07 vs 0.27±0.01 respectively, both p<0.01). Treatment with 20 mg/(kg·d) of losartan normalized BP(132±9 mmHg vs 178±17 mmHg,p<0.05) and HW/BW (0.32±0.03 vs 0.40±0.07,p<0.05). The lumen to media of mesenteric resistance arteries was decreased compared with control group (6.00±0.89 vs 8.96±1.23, p<0.01). Losartan normalized the lumen to media of hypertension group (8.87±1.25 vs 6.00±0.89,p<0.05). The p-ERK1/2 of aortas in hypertension group was overexpression compared with control group, and the overexpression of p-ERK1/2 was downregulated by treatment with losartan. Conclusion Losartan could regulate vascular remodeling in 2-kidney,1-cliped hypertensive rat by inhibiting the signaling pathway of ERK1/2.

Abstract:Aim To investigate the time-dependent changes in serum concentrations of endogenous inhibitor of nitric oxide synthase (NOS) asymmetric dimethylarginine (ADMA) in streptozotocin-induced diabetic rats and age-matched control rats, and to determine whether elevated endogenous ADMA is related to metabolic control and is implicated in the endothelium-dependent vasodilation dysfunction of diabetic rats at different stage of diabetic duration. Methods Serum levels of ADMA and L-arginine were measured by high-performance liquid chromatography at stages of 2-, 4- and 8-week diabetic duration. Thoracic aortic rings from diabetic rats with different diabetic duration and their age-matched control rats were studied for endothelium-dependent relaxation response to acetylcholine in vitro. Serum concentrations of glucose, glycosylated serum protein, and malondialdehyde, derived from lipid peroxidation were also examined to estimate metabolic control. Results Serum levels of ADMA significantly elevated in diabetic rats with 2-, 4-, and 8-week diabetic duration compared with their age-matched control rats(3.71±0.44 μmol/L vs 1.04±0.46 μmol/L for 2-week,3.54±1.70 μmol/L vs 0.95 ±0.13 μmol/L for 4-week,3.21±1.13 μmol/L vs 1.03±0.20 μmol/L for 8-week,n=5～6,all p<0.01), whereas serum levels of L-arginine were not different between control and diabetic rats at stages of 2-, 4-, and 8-week diabetic duration. Accordingly, the ratio of L-arginine and ADMA in diabetic rats was lower than that in their age-matched control rats. This elevation of ADMA was accompanied by impairment of relaxation response to acetylcholine of aortic rings in diabetic rats. Serum levels of glucose, glycosylated serum protein, and malondialdehyde were significantly increased in parallel with the elevation of ADMA in diabetic rats with 2-, 4-, and 8-week diabetic duration compared with their age-matched control rats. Conclusion These results reveal that the extent of elevation in serum ADMA in streptozotocin-induced diabetic rats with 2-, 4-, and 8-week diabetic duration is not correlated with the length of diabetic duration. This elevation of endogenous inhibitor of NOS in diabetic rats may relate to metabolic control of the disease and may be impli-cated in the endothelium-dependent vasodilation dysfunction associated with diabetes.

Abstract:Aim To investigate the time-dependent changes in serum concentrations of endogenous inhibitor of nitric oxide synthase (NOS) asymmetric dimethylarginine (ADMA) in streptozotocin-induced diabetic rats and age-matched control rats, and to determine whether elevated endogenous ADMA is related to metabolic control and is implicated in the endothelium-dependent vasodilation dysfunction of diabetic rats at different stage of diabetic duration. Methods Serum levels of ADMA and L-arginine were measured by high-performance liquid chromatography at stages of 2-, 4- and 8-week diabetic duration. Thoracic aortic rings from diabetic rats with different diabetic duration and their age-matched control rats were studied for endothelium-dependent relaxation response to acetylcholine in vitro. Serum concentrations of glucose, glycosylated serum protein, and malondialdehyde, derived from lipid peroxidation were also examined to estimate metabolic control. Results Serum levels of ADMA significantly elevated in diabetic rats with 2-, 4-, and 8-week diabetic duration compared with their age-matched control rats(3.71±0.44 μmol/L vs 1.04±0.46 μmol/L for 2-week,3.54±1.70 μmol/L vs 0.95 ±0.13 μmol/L for 4-week,3.21±1.13 μmol/L vs 1.03±0.20 μmol/L for 8-week,n=5～6,all p<0.01), whereas serum levels of L-arginine were not different between control and diabetic rats at stages of 2-, 4-, and 8-week diabetic duration. Accordingly, the ratio of L-arginine and ADMA in diabetic rats was lower than that in their age-matched control rats. This elevation of ADMA was accompanied by impairment of relaxation response to acetylcholine of aortic rings in diabetic rats. Serum levels of glucose, glycosylated serum protein, and malondialdehyde were significantly increased in parallel with the elevation of ADMA in diabetic rats with 2-, 4-, and 8-week diabetic duration compared with their age-matched control rats. Conclusion These results reveal that the extent of elevation in serum ADMA in streptozotocin-induced diabetic rats with 2-, 4-, and 8-week diabetic duration is not correlated with the length of diabetic duration. This elevation of endogenous inhibitor of NOS in diabetic rats may relate to metabolic control of the disease and may be impli-cated in the endothelium-dependent vasodilation dysfunction associated with diabetes.

Abstract:Aim To develop a novel method to effectively deliver enhanced green fluorescence protein (EGFP)C3 into Chinese hamster orary (CHO) cells in vitro by ultrasound-mediated microbubble destruction. Methods Expression of the EGFPC3 gene was quantified by laser confocal microscopy and FACS analysis. Cell viability was assayed by Trypan Blue staining. Results Ultrasound combined with microbubbles can enhance gene transfer in cultured cells, but the effect is vary according to the utrasound condition and concetration of microbubble. Optimal gene expression occurred with microbubble at the concentration of 10% and ultrasound parameter at 0.8 MHz, 1.0 W/cm 2,10% duty cycle, 60 s. Under this condition, the transfection rate of EGFPC3 gene with ultrasound-mediated microbubble destruction method was similar to that of transfection with lipofectamine (56.2%±2.6% versus 60.8%±4.1%, p>0.05) and the relative fluorescence intensity of EGFPC3 gene was as high as that of transfection with lipofectamine (2035±32 versus 2140±28, p>0.05). Furthermore both albumin microbubble and ultrasound had no effect on cell viability. The cell viablity were 94.1%±4.6% or 93.8%±3.1% when cultured with 10% albumin microbubble or under ultrasound at 0.8 MHz, 1.0 W/cm 2, 10% duty cycle, 60 s respectively, which were no significantly difference compared with controls. Conclusions These results suggest a possible new strategy for ultrasound-mediated microbubble destruction method in gene therapy.

Abstract:Aim To develop a novel method to effectively deliver enhanced green fluorescence protein (EGFP)C3 into Chinese hamster orary (CHO) cells in vitro by ultrasound-mediated microbubble destruction. Methods Expression of the EGFPC3 gene was quantified by laser confocal microscopy and FACS analysis. Cell viability was assayed by Trypan Blue staining. Results Ultrasound combined with microbubbles can enhance gene transfer in cultured cells, but the effect is vary according to the utrasound condition and concetration of microbubble. Optimal gene expression occurred with microbubble at the concentration of 10% and ultrasound parameter at 0.8 MHz, 1.0 W/cm 2,10% duty cycle, 60 s. Under this condition, the transfection rate of EGFPC3 gene with ultrasound-mediated microbubble destruction method was similar to that of transfection with lipofectamine (56.2%±2.6% versus 60.8%±4.1%, p>0.05) and the relative fluorescence intensity of EGFPC3 gene was as high as that of transfection with lipofectamine (2035±32 versus 2140±28, p>0.05). Furthermore both albumin microbubble and ultrasound had no effect on cell viability. The cell viablity were 94.1%±4.6% or 93.8%±3.1% when cultured with 10% albumin microbubble or under ultrasound at 0.8 MHz, 1.0 W/cm 2, 10% duty cycle, 60 s respectively, which were no significantly difference compared with controls. Conclusions These results suggest a possible new strategy for ultrasound-mediated microbubble destruction method in gene therapy.

Abstract:Aim To investigate the protective effect and probable mechanisms of Lumbrokinase on cerebral ischemia. Methods Using a mouse model of acute cerebral ischemia by ligating bilateral arteria carotis communis, the effect of Lumbrokinase on survival time was observed. The rat model of middle cerebral artery occlusion (MCAO) was established to measure infarction area, water content, nitric oxide (NO) content, nitric oxide synthase (NOS) activity and total-antioxidation capability (T-AOC) of brain after 24 h MCAO. Results Lumbrokinase 4 mg/kg significantly prolonged the survival time of mice (67.8±28.4 min) compared with that of the control group mice (9.5±4.4 min) (p<0.01). The infarction area obviously decreased and the reduced T-AOC caused by ischemia was improved in all Lumbrokinase groups (p<0.05 or p<0.01). The 3～6 mg/kg group showed decrease in cerebral water content(p<0.05). Lumbrokinase 6 mg/kg could also decrease NO content and NOS activity to 2.5±0.4 μmol/g and 718±68 U/g respectively (p<0.05). Conclusions The results indicated that Lumbrokinase plays a protective role on cerebral ischemia by decreasing infarction area and water content. The decrease of NO content and NOS activity and the improved T-AOC after ischemia may all contribute to the protective role.

Abstract:Aim To investigate the protective effect and probable mechanisms of Lumbrokinase on cerebral ischemia. Methods Using a mouse model of acute cerebral ischemia by ligating bilateral arteria carotis communis, the effect of Lumbrokinase on survival time was observed. The rat model of middle cerebral artery occlusion (MCAO) was established to measure infarction area, water content, nitric oxide (NO) content, nitric oxide synthase (NOS) activity and total-antioxidation capability (T-AOC) of brain after 24 h MCAO. Results Lumbrokinase 4 mg/kg significantly prolonged the survival time of mice (67.8±28.4 min) compared with that of the control group mice (9.5±4.4 min) (p<0.01). The infarction area obviously decreased and the reduced T-AOC caused by ischemia was improved in all Lumbrokinase groups (p<0.05 or p<0.01). The 3～6 mg/kg group showed decrease in cerebral water content(p<0.05). Lumbrokinase 6 mg/kg could also decrease NO content and NOS activity to 2.5±0.4 μmol/g and 718±68 U/g respectively (p<0.05). Conclusions The results indicated that Lumbrokinase plays a protective role on cerebral ischemia by decreasing infarction area and water content. The decrease of NO content and NOS activity and the improved T-AOC after ischemia may all contribute to the protective role.

Abstract:Aim To observe the effects of angiotensinⅡ type-1 receptor antagonist losartan on the neointimal proliferation and expression of matrix metalloproteinase-2(MMP-2) and tissue inhibitor of metalloproteinase-2(TIMP-2) after balloon angioplasty in Rabbits. Methods Twenty four rabbits were randomly divided into three groups: control group, model group and Losartan group. The rabbits of the latter two groups were fed with a 1.5% cholesterol diet starting one week before balloon injury. Then they were underwent ballon angioplasty in the celiac artery after the nine weeks of cholesterol diet. The losartan group was orally given losartan(10 mg/kg per day ) starting one week before angioplasty. Then the arteries were prepared for histological observation. Also, the sections were stained for MMP-2 and TIMP-2 immunohistochemistrical analysis. Results We found that after nine weeks of high-cholesterol diet, the serum total cholesterol and low density lipoprotein cholesterol levels of the model, losartan groups were significantly increased (p<0.01), and there were no changes in blood lipid parameters after losartan treatment. At the end of 13 weeks, compared with the model group, the intimal area (IA), intimal thickness (IT), IT/MT ratio (MT:media thickness),IA/MA ratio (MA:media area) of the losartan group were significantly decreased about 65%, 34%, 32%, 28%, respectively (p<0.01). Immunohistochemistrical analysis showed that there were no significant changes in the expression of MMP-2 after losartan treatment, while the expression of TIMP-2 significantly lowered, and the ratio of MMP-2/TIMP-2 significantly increased as a result of losartan treatment. Conclusion Losartan could significantly inhibit intimal proliferation during the restenosis after balloon injury without affecting blood lipid level; The mechanism of anti-resterosis by losartan may be related to inhibition of TIMP-2 and the revertion of balance of MMP-2/TIMP-2.

Abstract:Aim To observe the effects of angiotensinⅡ type-1 receptor antagonist losartan on the neointimal proliferation and expression of matrix metalloproteinase-2(MMP-2) and tissue inhibitor of metalloproteinase-2(TIMP-2) after balloon angioplasty in Rabbits. Methods Twenty four rabbits were randomly divided into three groups: control group, model group and Losartan group. The rabbits of the latter two groups were fed with a 1.5% cholesterol diet starting one week before balloon injury. Then they were underwent ballon angioplasty in the celiac artery after the nine weeks of cholesterol diet. The losartan group was orally given losartan(10 mg/kg per day ) starting one week before angioplasty. Then the arteries were prepared for histological observation. Also, the sections were stained for MMP-2 and TIMP-2 immunohistochemistrical analysis. Results We found that after nine weeks of high-cholesterol diet, the serum total cholesterol and low density lipoprotein cholesterol levels of the model, losartan groups were significantly increased (p<0.01), and there were no changes in blood lipid parameters after losartan treatment. At the end of 13 weeks, compared with the model group, the intimal area (IA), intimal thickness (IT), IT/MT ratio (MT:media thickness),IA/MA ratio (MA:media area) of the losartan group were significantly decreased about 65%, 34%, 32%, 28%, respectively (p<0.01). Immunohistochemistrical analysis showed that there were no significant changes in the expression of MMP-2 after losartan treatment, while the expression of TIMP-2 significantly lowered, and the ratio of MMP-2/TIMP-2 significantly increased as a result of losartan treatment. Conclusion Losartan could significantly inhibit intimal proliferation during the restenosis after balloon injury without affecting blood lipid level; The mechanism of anti-resterosis by losartan may be related to inhibition of TIMP-2 and the revertion of balance of MMP-2/TIMP-2.

Abstract:Aim To investigate the effect of Shuxin Yimai Jian (SXYMJ) on the apoptosis and the related gene Fas antigen and Fas ligand (FasL) of vascular balloon injury in thoracic aorta in rats with stagnancy of qi and blood stasis. Methods Fifty-eight rats were randomly divided into the sham group (n=10), SXYMJ group (n=24) and the control group (n=24). The thoracic aorta endothelium was denudated in the SXYMJ and control groups. SXYMJ was given from 7 days before operation to 14 days after operation in the SXYMJ group. Fas and FasL were detected by immunohistochemistry and the apoptotic cells were stained in situ by terminal deoxynucleotidyl transferase mediated-deoxyuridine triphosphate nick end labeling. Results Apoptotic cells could be detected in the smooth muscle cell (SMC) at the 14th day after operation. The expression of Fas and FasL in the neointima was increased by SXYMJ. The apoptosis of SMC was enhanced by SXYMJ, while the neointima area was attenuated. Conclusion SXYMJ probably regulates the apoptosis of SMC through Fas system and then inhibits the neointima hyperplasia.

Abstract:Aim To investigate the effect of Shuxin Yimai Jian (SXYMJ) on the apoptosis and the related gene Fas antigen and Fas ligand (FasL) of vascular balloon injury in thoracic aorta in rats with stagnancy of qi and blood stasis. Methods Fifty-eight rats were randomly divided into the sham group (n=10), SXYMJ group (n=24) and the control group (n=24). The thoracic aorta endothelium was denudated in the SXYMJ and control groups. SXYMJ was given from 7 days before operation to 14 days after operation in the SXYMJ group. Fas and FasL were detected by immunohistochemistry and the apoptotic cells were stained in situ by terminal deoxynucleotidyl transferase mediated-deoxyuridine triphosphate nick end labeling. Results Apoptotic cells could be detected in the smooth muscle cell (SMC) at the 14th day after operation. The expression of Fas and FasL in the neointima was increased by SXYMJ. The apoptosis of SMC was enhanced by SXYMJ, while the neointima area was attenuated. Conclusion SXYMJ probably regulates the apoptosis of SMC through Fas system and then inhibits the neointima hyperplasia.

Abstract:Aim To study the change of tumor necrosis factor (TNF)? interleukin-6 (IL-6)? interleukin-8 (IL-8) in cerebrospinal fluid (CSF) in patients with cerebral infarction. Methods The level of TNF? IL-6? IL-8 in CSF in patients with cerebral infarction in the period<1 week and >4 week and in twenty heathy elderly persons as control were measured by ELISA respectively. Results The level of TNF? IL-6? IL-8 in CSF in patients with cerebral infarction was higher than normal control. The degree of raise about TNF? IL-8 in period<1 week was more significant than>4 week. The level of IL-6 had no relation with the stage of disease. And there was a positive correlation between the increasing degree of TNF? IL-8 and the degree of neurological deficits. Conclusion The measure of level of TNF? IL-8? IL-6 in CSF could be taken as index of prognosis of cerebral infarction.

Abstract:Aim To study the change of tumor necrosis factor (TNF)? interleukin-6 (IL-6)? interleukin-8 (IL-8) in cerebrospinal fluid (CSF) in patients with cerebral infarction. Methods The level of TNF? IL-6? IL-8 in CSF in patients with cerebral infarction in the period<1 week and >4 week and in twenty heathy elderly persons as control were measured by ELISA respectively. Results The level of TNF? IL-6? IL-8 in CSF in patients with cerebral infarction was higher than normal control. The degree of raise about TNF? IL-8 in period<1 week was more significant than>4 week. The level of IL-6 had no relation with the stage of disease. And there was a positive correlation between the increasing degree of TNF? IL-8 and the degree of neurological deficits. Conclusion The measure of level of TNF? IL-8? IL-6 in CSF could be taken as index of prognosis of cerebral infarction.

Abstract:Aim To investigate the association between vascular endothelial growth factor (VEGF) and the severity of coronary lesions as well as coronary collateral development in patients with coronary heart disease. Methods The concentration of VEGF was measured by enzyme linked immunoserbent assay (ELISA) in 102 patients with angiographically documented coronary heart disease and 43 normal persons without any angiographically detectable coronary artery disease. The coronary artery score was recorded according to Leaman and the coronary collateral class was made according to Rentrop. The relationships between plasma VEGF and Leaman coronary artery score as well as coronary collateral circulation were assessed. Results The level of coronary artery plasma VEGF was obviously higher in the coronary disease group than in the control group (225±147 ng/L vs 74±52 ng/L ,p<0.01), and the mean VEGF was higher in coronary disease patients with collateral circulation than in patients without collateral circulation (299±152 ng/L vs 202±122 ng/L, p<0.01). The concentration of plasma VEGF was positively related to the Leaman score (r=0.693, p<0.001). Conclusions The plasma VEGF had correlation to both the severity of coronary stenosis and coronary collateral circulation in patients with coronary heart disease. VEGF might enhance collateral circulation development and doubly regulate the development of atherosclersis.

Abstract:Aim To investigate the association between vascular endothelial growth factor (VEGF) and the severity of coronary lesions as well as coronary collateral development in patients with coronary heart disease. Methods The concentration of VEGF was measured by enzyme linked immunoserbent assay (ELISA) in 102 patients with angiographically documented coronary heart disease and 43 normal persons without any angiographically detectable coronary artery disease. The coronary artery score was recorded according to Leaman and the coronary collateral class was made according to Rentrop. The relationships between plasma VEGF and Leaman coronary artery score as well as coronary collateral circulation were assessed. Results The level of coronary artery plasma VEGF was obviously higher in the coronary disease group than in the control group (225±147 ng/L vs 74±52 ng/L ,p<0.01), and the mean VEGF was higher in coronary disease patients with collateral circulation than in patients without collateral circulation (299±152 ng/L vs 202±122 ng/L, p<0.01). The concentration of plasma VEGF was positively related to the Leaman score (r=0.693, p<0.001). Conclusions The plasma VEGF had correlation to both the severity of coronary stenosis and coronary collateral circulation in patients with coronary heart disease. VEGF might enhance collateral circulation development and doubly regulate the development of atherosclersis.

Abstract:Aim To evaluate the clinic value of coronary artery calcification (CAC) detected by EBCT in adult patients with valvular heart disease (VHD). Methods 208 patients including 110 men and 98 women with age ranging from 42～75 years. All patients underwent EBCT examination and coronary angiography within 1 week before surgery. An Imatron C-150 EBCT scanner with scanning protocol of ECG-triggered single-slice mode was used to detect coronary calcification .The quantitative study of coronary calcification was carried out by Agatston method. Selected coronary agiography was performed by the Judkins technique. The five classic risk factors of coronary artery disease (CAD) were analyzed including diabetes mellitus? hypertension? hyperlipemia? smoking and family history of CAD. Results In the 208 patients, no coronary calcification was found in 114 cases detected by EBCT. Among them, 109 cases had no CAD (95.6%, p<0.005) and only 5 cases associated with CAD confirmed by coronary angiography. In the five patients with CAD, 4 case had at least one CAD risk factor. In all of the 208 cases, 58 patients had neither CAC nor CAD risk factor. Among the 58 cases, only one patient had CAD demonstrated by coronary angiography (1.72%). Conclusions Based on this study, the adult patients with valvular heart disease have neither coronary calcification detected by EBCT nor CAD risk factor. The accuracy to exclude CAD is rather high, more than 95.6%. We recommend the patients with VHD taking EBCT examination to detect CAC before surgery, that will markedly reduce application of invasive coronary angiography.

Abstract:Aim To evaluate the clinic value of coronary artery calcification (CAC) detected by EBCT in adult patients with valvular heart disease (VHD). Methods 208 patients including 110 men and 98 women with age ranging from 42～75 years. All patients underwent EBCT examination and coronary angiography within 1 week before surgery. An Imatron C-150 EBCT scanner with scanning protocol of ECG-triggered single-slice mode was used to detect coronary calcification .The quantitative study of coronary calcification was carried out by Agatston method. Selected coronary agiography was performed by the Judkins technique. The five classic risk factors of coronary artery disease (CAD) were analyzed including diabetes mellitus? hypertension? hyperlipemia? smoking and family history of CAD. Results In the 208 patients, no coronary calcification was found in 114 cases detected by EBCT. Among them, 109 cases had no CAD (95.6%, p<0.005) and only 5 cases associated with CAD confirmed by coronary angiography. In the five patients with CAD, 4 case had at least one CAD risk factor. In all of the 208 cases, 58 patients had neither CAC nor CAD risk factor. Among the 58 cases, only one patient had CAD demonstrated by coronary angiography (1.72%). Conclusions Based on this study, the adult patients with valvular heart disease have neither coronary calcification detected by EBCT nor CAD risk factor. The accuracy to exclude CAD is rather high, more than 95.6%. We recommend the patients with VHD taking EBCT examination to detect CAC before surgery, that will markedly reduce application of invasive coronary angiography.

Abstract:Aim To investigate the role of postprandial triglyceride levels in atherosclerosis, we examined the correlation between postprandial triglyceride levels and the carotid and femoral arteries atherosclerosis in elderly patients with coronary heart disease (CHD). Methods Carotid intima-media thickness (IMT) and plaque in the carotid and femoral arteries were measured by ultrasonography in 47 elderly patients with CHD and 30 controls. Plasma triglyceride(TG),total cholesterol, high density lipoprotein cholesterol, low density lipoprotein cholesterol, Apo A, Apo B levels were measured after overnight fasting and 4 h after a meal. Results The fasting cholesterol and Apo B were significantly higher in CHD group than those of the control group (p<0.05). The postprandial triglyceride and low density lipoprotein cholesterol were significantly higher and the postprandial high density lipoprotein cholesterol was significantly lower in CHD group than those of the control group (p<0.05). The IMT of common carotid arteries and the plaque index of carotid and femoral arteries in CHD group were significantly higher than those of the control group (p<0.01). Common carotid IMT was significantly increased in CHD group compared with the control group. Postprandial triglyceride levels were correlated with fasting triglyceride levels. Fasting total cholesterol was correlated with the plaque index of carotid. Conclusion Postprandial hypertriglyceride despite normal fasting triglycerid levels may be an risk factor for atherosclerosis in elderly patient.

Abstract:Aim To investigate the role of postprandial triglyceride levels in atherosclerosis, we examined the correlation between postprandial triglyceride levels and the carotid and femoral arteries atherosclerosis in elderly patients with coronary heart disease (CHD). Methods Carotid intima-media thickness (IMT) and plaque in the carotid and femoral arteries were measured by ultrasonography in 47 elderly patients with CHD and 30 controls. Plasma triglyceride(TG),total cholesterol, high density lipoprotein cholesterol, low density lipoprotein cholesterol, Apo A, Apo B levels were measured after overnight fasting and 4 h after a meal. Results The fasting cholesterol and Apo B were significantly higher in CHD group than those of the control group (p<0.05). The postprandial triglyceride and low density lipoprotein cholesterol were significantly higher and the postprandial high density lipoprotein cholesterol was significantly lower in CHD group than those of the control group (p<0.05). The IMT of common carotid arteries and the plaque index of carotid and femoral arteries in CHD group were significantly higher than those of the control group (p<0.01). Common carotid IMT was significantly increased in CHD group compared with the control group. Postprandial triglyceride levels were correlated with fasting triglyceride levels. Fasting total cholesterol was correlated with the plaque index of carotid. Conclusion Postprandial hypertriglyceride despite normal fasting triglycerid levels may be an risk factor for atherosclerosis in elderly patient.

Abstract:Aim The evaluation of the clinical feasibility of disruption of intracoronary plaques was conducted by angioplasty with high intensity, low frequency ultrasound. Methods 30 cases of coronary heart disease with stenosis ≥75% in one blood vessel were divided into Ultrasonic dissolved group and PTCA group randomly. Comparison of stenosis degree and ST segment in routine ECG and exercise tests before and after the treatment were conducted. Results Ultrasound was found to have dissolved atherosclerotic plaques in 15 cases. The residual stenosis was 43.0%±15.5% after the treatment, representing a decrease of 43.5%±17.7% compared with that before the treatment (40.1%±15.3% vs 84.7%±4.7%,p<0.05). In 5 out of these 15 patients, the residual stenosis was less than 30%. ST segment in ECG was markedly lifted up after the treatment in 15 patients. There were remarkable differences between the time needed for exercise-induced angina pectoris, the time for exercise-induced decrease in ST segment for 0.1 mV and the maximal range of decrease in ST segment before and those after the treatment (p<0.01). The residual stenosis was significantly more severe in ultrasonic group compared with PTCA group(43.0±15.5% vs 20.9±2.9%, p<0.01). Conclusions Angioplasty with high intensity, low frequency ultrasound can be used as a new approach for treating serious stenosis and improving blood flow of the coronary artery.

Abstract:Aim The evaluation of the clinical feasibility of disruption of intracoronary plaques was conducted by angioplasty with high intensity, low frequency ultrasound. Methods 30 cases of coronary heart disease with stenosis ≥75% in one blood vessel were divided into Ultrasonic dissolved group and PTCA group randomly. Comparison of stenosis degree and ST segment in routine ECG and exercise tests before and after the treatment were conducted. Results Ultrasound was found to have dissolved atherosclerotic plaques in 15 cases. The residual stenosis was 43.0%±15.5% after the treatment, representing a decrease of 43.5%±17.7% compared with that before the treatment (40.1%±15.3% vs 84.7%±4.7%,p<0.05). In 5 out of these 15 patients, the residual stenosis was less than 30%. ST segment in ECG was markedly lifted up after the treatment in 15 patients. There were remarkable differences between the time needed for exercise-induced angina pectoris, the time for exercise-induced decrease in ST segment for 0.1 mV and the maximal range of decrease in ST segment before and those after the treatment (p<0.01). The residual stenosis was significantly more severe in ultrasonic group compared with PTCA group(43.0±15.5% vs 20.9±2.9%, p<0.01). Conclusions Angioplasty with high intensity, low frequency ultrasound can be used as a new approach for treating serious stenosis and improving blood flow of the coronary artery.

Abstract:Aim To investigate the value of the ratio of systolic blood pressure (rSBP) using treadmill exercise test in diagnosing coronary heart disease (CHD). Methods Ninety-nine patients (56 with CHD, 43 without CHD) underwent treadmill exercise test and selected coronary angiography. Data were analyzed and the sensitivity, specificity, accuracy of rSBP and traditional index were compared. Results The sensitivity, specificity and accuracy of the ratio of systolic blood pressure were 53.6%, 83.7% and 66.7% respectively, while those of the traditional index were 91.1%, 55.8% and 75.8% respectively; The sensitivity of rSBp>0.91 in diagnosing single vessel disease, double or multi-vessel disease increased gradually, lower than that of traditional index. Regression analysis showed a significant relationship between peak HR and pre-exercise HR (r=0.442, p<0.001)and between peak BP and pre-exercise BP (r=0.578, p<0.0001). Conclusions The ratio of systolic blood pressure in exercise test can predict coronary artery disease and its severity. Its sensitivity was lower than that of traditional index, but specificity was higher than it.

Abstract:Aim To investigate the value of the ratio of systolic blood pressure (rSBP) using treadmill exercise test in diagnosing coronary heart disease (CHD). Methods Ninety-nine patients (56 with CHD, 43 without CHD) underwent treadmill exercise test and selected coronary angiography. Data were analyzed and the sensitivity, specificity, accuracy of rSBP and traditional index were compared. Results The sensitivity, specificity and accuracy of the ratio of systolic blood pressure were 53.6%, 83.7% and 66.7% respectively, while those of the traditional index were 91.1%, 55.8% and 75.8% respectively; The sensitivity of rSBp>0.91 in diagnosing single vessel disease, double or multi-vessel disease increased gradually, lower than that of traditional index. Regression analysis showed a significant relationship between peak HR and pre-exercise HR (r=0.442, p<0.001)and between peak BP and pre-exercise BP (r=0.578, p<0.0001). Conclusions The ratio of systolic blood pressure in exercise test can predict coronary artery disease and its severity. Its sensitivity was lower than that of traditional index, but specificity was higher than it.

Abstract:Aim To investigate the role of atherosclerosis in transient ischemic attack (TIA). Methods The comparative study was applied by CDFI between 32 cases of TIA and 24 cases without history of TIA and stroke. In both groups, the stenosis extent of carotid artery, the site with the plaque, the conditions of the plaque surface and the echo inside it were evaluated. Results There was no significant difference between the TIA and the control groups (p>0.05), when the stenosis extent of carotid artery was less than 49%. But when that was more than 50%, there was significant difference between the two groups (p>0.05). The risk for TIA was as 4.3 times as that of control group (Or=4.3, the creditable range: 1.34～13.89). The percentage of the weak echo inside the plaque in TIA and control group were 49.8%±16.4% and 23.6%±8.4%, respectively. Conclusion The carotid atherosclerosis is an important risk factor of TIA, and the stenosis extent of carotid artery and the plaque with weak echo inside are closely related with TIA.

Abstract:Aim To investigate the role of atherosclerosis in transient ischemic attack (TIA). Methods The comparative study was applied by CDFI between 32 cases of TIA and 24 cases without history of TIA and stroke. In both groups, the stenosis extent of carotid artery, the site with the plaque, the conditions of the plaque surface and the echo inside it were evaluated. Results There was no significant difference between the TIA and the control groups (p>0.05), when the stenosis extent of carotid artery was less than 49%. But when that was more than 50%, there was significant difference between the two groups (p>0.05). The risk for TIA was as 4.3 times as that of control group (Or=4.3, the creditable range: 1.34～13.89). The percentage of the weak echo inside the plaque in TIA and control group were 49.8%±16.4% and 23.6%±8.4%, respectively. Conclusion The carotid atherosclerosis is an important risk factor of TIA, and the stenosis extent of carotid artery and the plaque with weak echo inside are closely related with TIA.

Abstract:Aim To explore the relationship of left-ventricle hypertrophy (LVH), ventricular arrhythmia and heart rate variability (HRV) in patients with essential hypertension(EH). Methods A total of 94 patients were divided into two groups: EH without LVH (control group, n=47), and EH with LVH (LVH group, n=47). The structure and function of hearts were observed through cardio B-ultrasound. The ventricular premature beats and the parameters of HRV in every time-span were observed through 24 hours holter monitoring electrocardiogram. Results Both of the incidence and grades of ventricular premature beats for 24 hours in LVH group are obviously higher than that in control group (P=0.000). The parameters of HRV were not significantly different between two groups (p>0.05). The independent risks of the grades of ventricular premature beats are the interventricular septum (IVS) thickness, the left ventricular end-diastolic dimension (LVDd) and age of patients (p<0.05, p<0.001, p<0.05; respectively). Conclusion The incidence of severe ventricular arrhythmia in EH patients with LVH raises obviously, but no correlativity exists between LVH and HRV. IVS, LVDd, and age are the independent risk factors of ventricular arrhythmia in EH patients.

Abstract:Aim To explore the relationship of left-ventricle hypertrophy (LVH), ventricular arrhythmia and heart rate variability (HRV) in patients with essential hypertension(EH). Methods A total of 94 patients were divided into two groups: EH without LVH (control group, n=47), and EH with LVH (LVH group, n=47). The structure and function of hearts were observed through cardio B-ultrasound. The ventricular premature beats and the parameters of HRV in every time-span were observed through 24 hours holter monitoring electrocardiogram. Results Both of the incidence and grades of ventricular premature beats for 24 hours in LVH group are obviously higher than that in control group (P=0.000). The parameters of HRV were not significantly different between two groups (p>0.05). The independent risks of the grades of ventricular premature beats are the interventricular septum (IVS) thickness, the left ventricular end-diastolic dimension (LVDd) and age of patients (p<0.05, p<0.001, p<0.05; respectively). Conclusion The incidence of severe ventricular arrhythmia in EH patients with LVH raises obviously, but no correlativity exists between LVH and HRV. IVS, LVDd, and age are the independent risk factors of ventricular arrhythmia in EH patients.

Abstract:Aim To explore the efficacy of defibrase in treatment of acute cerebral infarction and the effect of hemorheology. Methods 100 cases with acute cerebra infarction were randomly allocated to two groups: 50 cases in treatment group were given intravenous infusion of defibrase 10 u every other day for 3 times, and 50 cases in control group were given Co-Danshen 20 mL every day for 7 days. Then before and after treatment in 14 days, two groups were observed in time of taking effect, curative effect, side effect and hemorheological change respectively. Results The total effective rate of the treatment group was 90%, which was significantly higher than that of the control group (68%) (p<0.05), and the time taking effect was earlier (p<0.01). At the same time, indexes of hemorheology were remarkably declined (p<0.01 or p<0.05). There was only 1 case that came about bleeding cerebral infarction. Conclusion Defibrase is efficacious, safe and rapid for the treatment of Acute Cerebra Infarction.

Abstract:Aim To explore the efficacy of defibrase in treatment of acute cerebral infarction and the effect of hemorheology. Methods 100 cases with acute cerebra infarction were randomly allocated to two groups: 50 cases in treatment group were given intravenous infusion of defibrase 10 u every other day for 3 times, and 50 cases in control group were given Co-Danshen 20 mL every day for 7 days. Then before and after treatment in 14 days, two groups were observed in time of taking effect, curative effect, side effect and hemorheological change respectively. Results The total effective rate of the treatment group was 90%, which was significantly higher than that of the control group (68%) (p<0.05), and the time taking effect was earlier (p<0.01). At the same time, indexes of hemorheology were remarkably declined (p<0.01 or p<0.05). There was only 1 case that came about bleeding cerebral infarction. Conclusion Defibrase is efficacious, safe and rapid for the treatment of Acute Cerebra Infarction.