1.Quantitative detection of the expression level of transforming growth factor-β1 and-β2 in rat retina with real time PCR
International Eye Science 2008;8(6):1076-1078
AIM: To quantitatively detect the expression level of transforming growth factor-β1 (TGF-β1) and transforming growth factor-β2 (TGF-β2) genes in the retina of normal rat in order to determine the expression difference of TGF-β1 and TGF-β2 in retina.METHODS: The total RNA was isolated from which the first strand of cDNA was prepared. The mRNA levels of TGF-β1 and TGF-β2 were detected quantitatively by real time polymerase chain reaction (PCR).RESULTS: The mRNA levels of TGF-β1 and TGF-β2 were 0.0008±0.0003 and 0.0378±0.009, respectively. Expression of TGF-β2 was obviously higher than that of TGF-β1 in rat retina with statistical significance (t=12.37, P<0.001). The ratio of TGF-β2/TG-β1 was 55.00±26.61.CONCLUSION: QRT-PCR could specifically and accurately detect gene expression level in rat retina. In retina the TGF-β2 gene was expressed more abundantly than TGF-β1. It is suggested that TGF-β2 play an important role in retina diseases.
2.Expression of transforming growth factor-β type Ⅰ receptor and transforming growth factor-β type Ⅱ receptor in rat retina
International Eye Science 2008;8(6):1073-1075
AIM: To quantitatively investigate transforming growth factor-β type Ⅰ receptor (TβRⅠ) and transforming growth factor-β type Ⅱ receptor (TβRⅡ) gene expressions in rat retina.METHODS: Sprague-Dawley rats were chosen in this research. Gene expression was detected quantitatively by reverse transcription polymerase chain reaction (RT-PCR) analysis. RESULTS: The expression level of TβRⅠ and TβRⅡ were 0.00034±0.00013 and 0.0001±0.00005, respectively. The expression level of TβRⅠ was obviously higher than that of TβRⅡ in the rat retina with statistical significance (P<0.01). The ratio of TβRⅠ/TβRⅡ was 3.9±1.7.CONCLUSION: Real time quantitative RT-PCR is an effective method to detect differential expression genes in retina. The change of TβRⅠ/TβRⅡ expression may play an important role in the pathogenesis of retinopathy, which could be further investigated in its significance in the development of proliferation retinopathy.
3.Gene expression of transforming growth factor-β2 in retina of diabetic rats
International Eye Science 2008;8(6):1065-1069
AIM: To detect the gene expression of TGF-β2 in retinas of diabetic rats at different stages, to observe and analyze the effect of TGF-β2 on the retinas of diabetic rats, to explore the role of TGF-β2 in pathogenesis of diabetic retinopathy (DR), and to provide experiment data and experience for further clinic studies.METHODS: Sprague-Dawley (SD) rats were used and retinas were dissected. The total RNA was isolated from which the first strand of cDNA was prepared. Diabetes mellitus was induced by a single intraperitoneal injection of 60mg/kg streptozotocin (STZ) and the rats were held without insulin treatment until sacrifice. Besides, agematched rats treated with saline were used as controls. Tail vein blood glucose was measured after 2 days and rats were considered hyperglycemic if blood glucose reading>16.7mmol/L. Animals with blood glucose level<16.7mmol/L were excluded from the study. The rats were killed at the 4th, 8th, 12th, 16th, 20th and 24th week respectively after hyperglycemic models were established. The retinas were separated and preserved in liquid nitrogen. The expressions of TGF-β2 gene mRNA were detected by reverse transcription PCR (RT-PCR).RESULTS: The RNA of rat retina was integrative enough to be used to further carry out PCR analysis. Compared with control groups, the expression of TGF-β2 mRNA in retinas of diabetic rats was up-regulated at the 4th week, but there was no statistical difference (P>0.05); it was down-regulated at the 8th week, and there was statistical difference (P<0.05); it was also down-regulated at the 12th week, and there was statistical difference (P<0.05); at the 16th week there was no statistical difference (P>0.05); it was up-regulated at the 20th week, but there was no statistical difference (P>0.05); it continued to be up-regulated at the 24th week, and there was statistical difference (P<0.05).CONCLUSION: Since the expression of TGF-β2 mRNA in retinas of diabetic rats was down-regulated at the 8th week and 12th week statistically, up-regulated at the 24th week statistically, it has obviously shown that TGF-β2 was down-and up-regulated through the period of DR. That is, its changes are diphasic with time. It may confirm that TGF-β2, with the characteristic of diphasic regulation, played an important role in DR. It is necessary to study it furthermore.
4.Expression of transforming growth factor-β type 1 receptor and transforming growth factor type 2 receptor in diabetic rat retina
Dan, CAO ; Lin, LIU ; Wei, SHEN
International Eye Science 2006;6(2):247-251
· AIM: To quantitatively detect gene expression level of transforming growth factor-β type 1 receptor (Tβ R 1) and transforming growth factor-β type 2 receptor (Tβ R 2) in different stage of diabetic rats' retina. To observe and analyze the effect of transforming growth factor-βreceptors on the retina of rat diabetic animal model.· METHODS: 28 healthy adult Sprague-Dawley rats were chosen and randomly divided into two groups of normal control (CON) and diabetes mellitus (DM). Diabetes was induced by streptozotocin (STZ) intraperitoneal injection.Gene expression was detected quantitatively with real-time fluorescence quantitative reverse transcription polymerase chain reaction (QRT-PCR).· RESULTS: The mRNA level of Tβ R 1 and Tβ R 2 was 0.000493± 0.000133 and 0.000166± 0.000057 at 4wk. The mRNA level of Tβ R 1 and Tβ R 2 was 0.000608± 0.000232 and 0.000113± 0.000049 at 12wk. Tβ R 1 expression was gradually elevated during the progression of diabetic retinopathy, Tβ R 2 expression was up-regulated at 4wk,but down-regulated at 12wk.Tβ R 2) may play important role in the pathogenesis of diabetic retinopathy.
5.Experimental study of grape seed proanthocyanidin extract plus atorvastatin in the treatment of atherosclerosis
Lin SHEN ; Haiqing GAO ; Xiangju LIU
Chinese Journal of Geriatrics 2009;28(10):856-859
Objective To observe the effects of grape seed proanthocyanidin extract(GSPE) plus atorvastatin treatment on matrix metalloproteinase-9 ( MMP-9) in experimental atherosclerosis rabbits and to explore its mechanism. Methods Thirty male New Zealand rabbits were randomized into five groups. The normal control group were fed with standard diet for 24 weeks. And the other groups were fed with standard diet containing 1 % cholesterol for 12 weeks. In the sequential 12 weeks, the model control group was fed with standard diet. The GSPE group was fed with standard diet containing 1% grape seed proanthocyanidin(GSP). The atorvastatin group was fed with standard diet containing atorvastatin(2. 5 mg · kg~(-1) · d~(-1)). The GSPE plus atorvastatin group was fed with standard diet containing 1% GSP and atorvastatin (2. 5 mg~(-1) · kg~(-1) · d~(-1)). Blood samples were drawn from ear middle arteries of rabbits just before the experiment and at the 12th and the 24th weekend of the experiment. All the rabbits were fasted for at least eight hours before the blood was drawn. The blood samples were analyzed for the matrix metalloproteinase-9 (MMP-9). All the rabbits were sacrificed at the 24th weekend, and the expression of MMP-9 was observed in the thoracic aortic tissue using immunohistochemistry technique. Results The serum level and aorta expression of MMP-9 were increased in model group compared to control group (all P< 0. 05). The severity of atherosclerosis was less in three drug groups than that in model control group. The GSPE,atorvastatin and GSPE plus atorvastatin groups versus model group showed less atherosclerotic lession, the decreased expression of aorta MMP-9 and the decreased serum level of MMP-9 C(1. 06±0. 21), ( 1. 07 ±0.20), (0.81 + 0.16) vs. (1. 32±0. 24)ng/ml,all P<0. 05]. The effect in the GSPE and atorvastatin group was most obvious. Conclusions GSPE plus atorvastatin group has the most efficacy of anti-atherogenesis, which is associated with its reducing the expression of matrix metalloproteinase.
6.Clinical features and nursing for the male elderly patients with peripheral atherosclerosis
Lin SHEN ; Zhiying LIU ; Jian CAO
Chinese Journal of Practical Nursing 2009;25(5):19-21
Objective To investigate the clinical characteristics of peripheral arteriosclerosis in elderly males and to discuss corresponding nursing strategies. Methods 65 patients were divided into the peripheral arteriosclerosis group (the experimental group, 38 patients) and the nonperipheral arteriosclerosis group (the control group,27 patients) according to echocardiographic diagnosis. Questionnaires were designed to record factors such as age, smoking history, alcohol consumption history, primary hypertension, diabetes mellitus, coronary heart disease, blood pressure, height and body weight. Body mass index (BMI) was calculated. Results Hypertension course was much longer and proportion of patients with hypercholesterolemia and coronary heart disease and smoking index, TG, LDL-C and HDL-C levels were significantly higher in the experimental group than those in the control group. Conclusions Nursing care should be carried out from psychological, life style, medication and health education perspectives in elderly males with peripheral arteriosclerosis, aiming at improving their quality of life.
7.Effects of narcotics on adenyl cyclase and phosphodiesterase activity in rabbit bronchial smooth muscle.
Cunming LIU ; Jianfan SHEN ; Guifang LIN
Chinese Journal of Anesthesiology 1994;0(05):-
The effects of morphine, pethidine and fentanyl on adenyl cyclase (AC) and phospho- diesterases (PDE) in rabbit bronchial smooth muscle were examined in vitro. The results were as follows: morphine could inhibit the activity of AC which was decreased from 15. 9911 ? 0. 8597u to 15.0463?0. 7989u, P
8.Effects of propofol combined with resveratrol pretreatment on hepatic ischemia-reperfusion injury in rats
Xin SHEN ; Ge ZHAO ; Tingting LIU ; Rui WANG ; Lin LIU
Chinese Journal of Anesthesiology 2013;(3):371-375
Objective To evaluate the effects of propofol combined with resveratrol on hepatic ischemiareperfusion (I/R) injury in rats.Methods One hundred and eight male Sprague-Dawley rats,aged 6-7 weeks,weighing 220-270 g,were randomly divided into 6 groups (n =18 each):sham operation group (group S) ; I/R group; solvent group (group TW-80) ; propofol group (group P) ; resveratrol group (group R) ; propofol combined with resveratrol group (group P + R).The animals were anesthetized with intraperitoneal 10 % chloral hydrate 3.5-4.0 ml/kg.Liver ischemia was produced by clamping the first hepatic portal for 30 min,followed by 12 h reperfusion.In group P,propofol was infused intravenously at 10 mg· kg-1 ·h-1 starting from 10 min before ischemia until the end of operation.In group R,resveratrol 10 mg/kg was injected intravenously at 10 min before ischemia.In group P + R,resveratrol 10 mg/kg was injected intravenously and then propofol was infused intravenously at 10 mg· kg-1· h-1 starting from 10 min before ischemia until the end of operation.The equal volume of normal saline was given instead in groups S and I/R,and the equal volume of TW-80 was given instead in group TW-80.Six rats in each group were chosen at 3,6 and 12 h of reperfusion and blood samples were taken from the superior and inferior vena cava for measurement of serum alanine aminotransferase (ALT) and aspartate amino transferase (AST) activities.The rats were then sacrificed and livers were removed for microscopic examination and for determination of superoxide dismutase (SOD) and myeloperoxidase (MPO) activities,inducible nitric oxide synthase (iNOS) expression,and malondialdehyde (MDA) and nitric oxide (NO) contents in liver tissues.Results Compared with group S,the serum ALT and AST activities,and MDA and NO contents,MPO activity and iNOS expression in liver tissues were significantly increased,and the activities of SOD were decreased in the other four groups (P <0.05).Compared with group I/R,the serum ALT and AST activities,and MDA and NO contents,MPO activity and iNOS expression in liver tissues were significantly decreased,and the activities of SOD were increased in groups P,R and P + R (P < 0.05).Compared with groups P and R,SOD activity was significantly increased,and the other parameters were decreased in group P + R (P < 0.05).The pathological changes were significantly attenuated in groups P,R and P + R compared with group I/R.Conclusion Pretreatment with propofol and resveratrol can attenuate hepatic I/R injury in rats by increasing antioxidation and inhibiting lipid peroxidation and reducing production of endogenous NO,and the combination of the two agents provides better efficacy than etheir alone.
9.The correlation between carotid and femoral pulse wave velocity and composition of the ascending aorta in the patients with coronary disease
Beian YOU ; Haiqing GAO ; Lin SHEN ; Xiangju LIU ; Jie QIU
Chinese Journal of Geriatrics 2012;31(9):758-761
Objective To investigate the association between carotid-femoral pulse wave velocity (cfPWV) and composition of the ascending aorta in patients with coronary heart disease (CAD),Methods The study population comprised 60 consecutive CAD patients who underwent coronary artery bypass graft (CABG) surgery in Qilu Hospital of Shandong University.cfPWV vas measured using an automatic device (Complior,Artech,France).A quantitative study was conducted on ascending aorta specimens by histological observation (Masson staining and weigert's resorcin-fuchsin staining) and computer image analysis.Bivariate analyses were performed to study the association between composition of the ascending aortic media and cfPWV.Results cfPWV of the CAD patients was higher (14.2±2.0) m/s than that of the normal subjects.On Masson's-stained specimen slides,disorganization of smooth muscle and focal accumulations of collagen (44.1 ± 3.9)% were visible alongthe medial aorta of the CAD patients.Weigert's-stained cross sections of the ascending aortic media in CAD patients frequently exhibited focal breakdown or discontinuous segments of elastic fibers(18.4±3.2)%,cfPWV had a positive correlation with relative contents of collagen in the ascending aorta(r=0.68,P<0.01)and a negative correlation vith relative contents of elastin in the ascending aorta (r=-0.59,P<0.01),but no relation with relative contents of smooth muscle(r=0.01,P>0.05).Conclusions The reduced aortic elasticity in CAD patients can be partly ascribed to decreased elastin,increased collagen,and their disorganization.cfPWV can reflect the quantitative changes of collagen and elastin in the ascending aortic media in CAD patients.
10.Relationship between morphological features of carotid arterial atherosclerosis plaque and cardiovascular risk factors in the aged
Lin SHEN ; Haiqing GAO ; Beian YOU ; Jie QIU ; Xiangju LIU
Chinese Journal of Geriatrics 2011;30(4):265-268
Objective To explore the relationship between morphological features of carotid arterial atherosclerosis plaque and various cardiovascular risk factors in the aged.Methods The 321patients who underwent cerebral angiography were classified into two groups:256 patients with carotid arterial atherosclerosis and 65 patients without plaque.The relationships between carotid arterial plaque and cardiovascular risk factors including age,gender,cholesterol(TC),triglyceride (TG),high density lipoprotein cholesterol (HDL-C),low density lipoprotein cholesterol(LDL-C),C-reactive protein(CRP),homocysteine (HCY),hypertension,diabetes,smoking and drinking were analyzed.The relationships between morphological features,including the stenosis degree,surface fairing of plaque,and cardiovascular risk factors were analyzed.After 1 year follow-up,the prospective cardiovascular and cerebrovascular events were analyzed in the two groups.Results Carotid atherosclerosis was connected with cardiovascular risk factors such as TC,LDL-C,uric acid,CRP,hypertension,diabetes,smoking and drinking.Positive correlation was showed between the stenosis degree of carotid artery and age,smoking and diabetes.Closely correlation was showed between surface fairing of carotid arterial plaque and LDL-C,CRP,smoking and diabetes.The stenosis degree of carotid artery only showed positive correlation with cerebrovascular events,but the morphological features of surface fairing of carotid arterial plaque equally showed positive correlations with cerebrovascular events and cardiovascular events.Conclusions The development of carotid arterial atherosclerosis plaque in the aged is closely related to cardiovascular risk factors.Morphological features of carotid arterial plaque may respond the level of general atherosclerosis better than stenosis degree,and it is also in close correlation with cerebrovascular events and cardiovascular events.