1.Suppressive effect of adenovirus-mediated RNA interference on angiotensinⅡreceptor subtypes mRNA expression in mice brainstem
Chinese Journal of Geriatrics 2008;27(6):462-465
Objective To investigate the inhibitory effects of RNA silencing via adenovirusmediated angiotensin Ⅱ receptor subtypes shRNA(Ad-ATlaR-shRNA and Ad-ATlbR-shRNA)in mice brainstem nucleus tractus solitarius(NTS)in vivo. Methods C57BL mice were used as animal model.The microinjection into the nucleus of NTS was adopted.Two groups of male C57BL mice(n =7,n=8)were selected.After 10 days of microinjection,mice were killed and their brain tissue were fixed and sectioned.The levels of ATlaR mRNA and ATlbR mRNA at both sides of NTS were examined by in situ hybridization. Results The expression of ATlaR mRNA was significantly inhibited from 1.81/μCi/mg to 0.71μCi/mg[(61.6±6.8)%,P<0.01]by Ad-ATlaR-shRNA.Meanwhile,ATlbR mRNA expression was consistent at both sides,with no significant difference(P>0.05)after Ad-ATlaR-shRNA microinjection.ATlbR mRNA expression at the side with AdATlbR-shRNA injection was significantly inhibited from 10.28/μCi/mg to 4.97μCi/mg[(51.6±5.2)%,P<0.01]by Ad-ATlbR-shRNA.Meanwhile,ATlaR mRNA probe were consistent at both sides,with no significant difference(P>0.05). Conclusions The results show that both AdATlaR-shRNA and Ad-ATlbR-shRNA inhibit the corresponding receptor mRNA expression,and there is no cross-action for each other.
2.The dose-effect and its mechanisms of preventive effects of ~(103) Pd radioactive stent on in-stent restenosis in rabbits
Jiyan CHEN ; Hong YAN ; Shuguang LIN
Chinese Journal of Interventional Cardiology 2003;0(05):-
Objective To investigate the dose-response and its mechanisms of preventive effect of 103 Pd radioactive stent on restenosis in rabbits after balloon injury. Methods The abdominal arteries of the rabbits randomized into common or radioactive stent groups (including 5 Gy, 15 Gy, 25 Gy, 35 Gy groups) were balloon injured, and then stented. Intravascular ultrasound and angiography on abdominal arteries, immunohistochemical study of PCNA, and TUNEL assay were performed 8 weeks after stenting. Results Both the minimal lumen diameters and in-stent lumen areas in 103 Pd radioactive stent groups [(1.94?0.21) mm and (5.99?0.43) mm2, (2.17?0.12) mm and (7.01?0.93) mm2, (2.43?0.22) mm and (8.09?0.89) mm2, (2.56?0.27) mm and (8.49?0.54) mm2 in turn in 5 Gy, 15 Gy, 25 Gy, 35 Gy groups, respectively]were significantly improved with increases of the doses at the end of 8 weeks,revealing a dose-response effect. The positive ratio of PCNA in each 103 Pd stent group was reduced as compared with that in commen stent group (P
3.Oxidized low density lipoprotein induces the expression of macrophage migration inhibitory factor
Qiuxiong LIN ; Xiyong YU ; Zhixin SHAN ; Shuguang LIN ; Lan HUIYAO
Chinese Journal of Pathophysiology 2000;0(10):-
AIM: To examine expression of macrophage migration inhibitroy factor (MIF) gene and protein in macrophages induced by oxidized low density lipoprotein (ox-LDL). METHODS: Macrophages were incubated with ox-LDL at the concentration of 150 mg/L for time course (0-36 h) and with ox-LDL at the different concentrations (0-300 mg/L) for 24 h, expression of MIF mRNA and protein were detected by RT-PCR and ELISA. RESULTS: The results showed that ox-LDL increased MIF gene and protein expression in macrophages in a dose and time-dependent manner. After the exposure of macrophage to ox-LDL, the expression of MIF mRNA level increased consistently with protein. CONCLUSION: MIF may play an important role in atherosclerosis. [
4.Mechanism involved in the prevention of sub hypotensive dose of ramipril on hypertensive cardiac hypertrophy of rat
Luyuan CHEN ; Yanfang CHEN ; Shuguang LIN ; Zuojun LIN ;
Chinese Pharmacological Bulletin 1986;0(06):-
Aim To examine the role of antioxidant effect of subhypotensive dose of ACE inhibitor ramipril in its anti hypertrophic effect in rat myocardium. METHODS Model of hypertension induced myocardial hypertrophy of rat was reproduced by coarctation of abdominal aorta. The effects of nitric oxide (NO) synthase inhibitor N L arginine (6 mg?kg -1 ?d -1 ) and Vitamin E(200 U?d -1 ) on cardiac hypertrophy were examined. The change of superoxide dismutase (SOD) and NO production (assayed as NO 2 -) were monitored. RESULTS The myocardial hypertrophy was effectively prevented under hypertensive condition after treatment with ramipril for 8 week, while SOD activity and NO 2 - content were significantly increased and lipid peroxidation was significantly attenuated. Vitamin E enhanced ramipril effect and N L arginine reversed its benficail effects. CONCLUSION These data showed that sub hypotensive dose of ramipril and in combination with vitamin E enhances myocardial antioxidative capability and NO release, which plays an important role in the mechanism of action involved in their antihypertensive cardiac hypertrophic effects.
5.Effects of tetrandrine on hydroxyproline content and myosin ATPase activity of hypertrophied myocardium in renovascular hypertensive rats
Zean LU ; Qingping LI ; Manren RAO ; Xiyong YU ; Shuguang LIN
Chinese Journal of Pharmacology and Toxicology 2001;15(2):121-124
To study the effects of tetrandrine (Tet) on hypertrophied myocardial hydroxyproline content and myosin ATPase activity, left ventricular hypertrophy(LVH) was induced by renovascular hypertension (two-kidney, one-clip) in rats. Eight weeks after operation Tet 50 mg*kg-1*d-1 and enalapril(Ena) 6 mg*kg-1*d-1 were given by gavage for 8 weeks. The results showed that hydroxyproline content in LVH group was much higher than that of sham-operated one 〔(5.9±0.3) vs (3.6±0.4) mg*g-1 dry weight〕, and was decreased by 28.2% and 39.0% in Tet and Ena groups, respectively. Myosin ATPase activity in LVH group was much lower than that of sham-operated group 〔(0.43±0.09) vs (0.97±0.06) mmol Pi*min-1*g-1 protein, P<0.01〕. In Tet and Ena groups they were 60.5% and 118.6% higher than that of LVH group, respectively. The results suggest that Tet or Ena partially reduce the hydroxyproline content and elevate myosin ATPase activity of hypertrophied myocardium in renovascular hypertensive rats.
6.Survivin-siRNA inhibits proliferation of lung cancer A549 cells and enhances their chemosensitity to cisplatin
Shuguang ZHANG ; Xiaofan LIU ; Jiang DU ; Wenya LI ; Lin ZHANG
Chinese Journal of Cancer Biotherapy 2009;16(6):583-587
Objective:To investigate the effect of survivin-siRNA plasmid on survivin expression in human lung cancer cell line A549, and to observe its effect on the apoptosis, proliferation, and chemosensitivity of A549 cells. Methods: pSilencer-survivin-siRNA (survivin-siRNA) plasmid was constructed using pSilencer-U6 plasmid and was transfected into A549 cells. Expression of survivin mRNA and protein was examined by RT-PCR and Western blotting analysis, respec-tively. Apoptosis and proliferation of A549 cells were examined by DAPI staining and MTT, respectively. Results: Sur-vivin-siRNA plasmid was successfully constructed, and it significantly inhibited survivin mRNA and protein expression in A549 cells. Survivin-siRNA transfection induced apoptosis, inhibited proliferation and increased chemosensitivity of A549 cells to cisplatin. Conclusion: pSilencer-survivin-siRNA can silence survivin expression in A549 cells and subsequently inhibit proliferation, promote apoptosis, and enhance chemosensitivity of A549 cells to cisplatin. Survivin may serve as a potential target for gene therapy of lung cancer.
7.Oxided low density lipoproteins induce apoptosis with enhanced expression of Fas antigen in vascular endothelial cells
Ziwang HU ; Zhihua QUAN ; Yanfang CHEN ; Suizhan WANG ; Shuguang LIN
Chinese Journal of Pathophysiology 1989;0(05):-
AIM: To explore the effect of oxides of low density lipoprotein (OX - LDL)on apoptosis in vas- cular endothelial cells(EC) and its possible mechanisms. METHODS: Culted vascular ECS were incubated with OX - LDL(0.1?g/L or 0. 3?g/L) for 4 hours, flow cytometer with PI staining methed was used to determine the rate of cellular apoptosis, gel - electrophoresis was performed to observe foe DNA ladder of apoptosis, cellular RNA was isolated and reverse transcription-polymerase chain reaction (RT - PCR)was performed to determine Fas antigen ex- pression. RESULTS:OX - LDL treatment induced apoptosis in EC dose - dependently. No DNA ladder was ob- served in control EC. But it presented in EC beated with low dose of OX - LDL, and was even more when beted with higher dose of OX - LDL. RT - PCR revealed Fas antigen baldly had any expression in basic condition, OX - LDL treatment induced Fas antigen in EC in a dose dependent manner. CONCLUSION: OX - LDL can induce apoptosis in EC, upregulaton of Fas antigen expression maybe one of the possible mechanisms.
8.Transforming growth factor-? conjugated with cytotoxin saporin inhibits specifically proliferating vascular smooth muscle cells
Jun YANG ; Shuguang LIN ; Xiyong YU ; Qidong TANG ;
Chinese Pharmacological Bulletin 1987;0(02):-
AIM To testify the special cytotoxicity of TGF? SAP on proliferating vascular smooth muscle cells and endothelial cells. METHODS Conjugation of saporin to TGF? was accomplished after derivatization of saporin and TGF? with N succinimidyl 3 (2 pyridyldithio) proprionate. Cytotoxicity assays were measured by cell count. The studies of influence of TGF? SAP on values of thymidine and leucine incorporation into SMCs and ECs were measured by 3H thymidine uptake and 3H leucine uptake, respectively. RESULTS Cytotoxicity assays testified TGF? SAP conjugate could inhibit remarkably proliferation of SMCs in culture. The values of thymidine of TGF? SAP group (1?10 -9 mol?L -1 and 1?10 -7 mol?L -1 ) in comparison significantly decreased to 60 9% and 56 0% of the control group respectively, suggesting that cellular DNA synthesis obviously decreased as TGF? SAP was added. But saporin did not affect cellular DNA synthesis at higher level. The rate of 3H leucine incorporation of TGF? SAP group significantly decreased to 47 3% of the control group, suggesting that SMCs protein synthesis obviously decreased as TGF? SAP was added. But TGF? SAP at the same level did not affect DNA synthesis and protein synthesis of ECs compared with the control group. CONCLUSION TGF? SAP possesses the more effective cytotoxicity than saporin and the more specific citotoxicity on proliferating vascular smooth muscle cells than on proliferating endothelial cells.
9.A survey of dietary iodine intake among residents in Fujian Province
Zhihui CHEN ; Min HU ; Xi LIN ; Jiani WU ; Shuguang LIN ; Zhaohe LIN ; Muhua WANG
Chinese Journal of Endemiology 2014;(4):414-418
Objective To survey the dietary iodine intake among residents in the coastal province of Fujian after universal salt iodization. Methods Using multi-stage stratified random sampling method, four different geographic areas of coastal cities, inland cities, coastal rural and inland rural areas were selected; 3 counties (cities, districts) were selected from each area; 30 families were selected from each of those counties (cities, districts). Total dietary study method was applied to investigate the basic information, all food and water intake during the investigation, the spice variety, quantity and cooking methods, and recipes. All foods consumed were divided into 13 classes:grains and their products, beans(including nuts) and their products, potato and its products, meat and its products, eggs and their products, aquatic products, milk and dairy products, vegetables, fruits and their products, sugar and sugar products, beverage and water, alcohol and seaweeds. Food samples were collected and iodine contents of different types of food were measured, and daily food consumption of the reference man(adult male) in the four areas was calculated. Then the male adult dietary iodine intake in the four areas and Fujian Province was estimated. Results A total of 69 families(accounting for 19.2% of the 359 families surveyed) ate seaweed foods during the three days survey period. The average salt intakes per reference man were 4.5, 5.8, 7.5, 12.3 g, respectively, in coastal cities, inland cities, coastal rural and inland rural. The adult male dietary iodine intake in Fujian Province was 343.8μg/d, and the values were 234.4, 347.1, 328.4, 465.3μg/d, respectively, in coastal cities, coastal rural, inland cities and inland rural. If people did not eat seaweed foods, the results would be 239.7, 137.5, 253.6, 200.3, 367.4μg/d. The main sources of dietary iodine intake were vegetables (33.2%), seaweeds(30.3%), meat(13.4%) and aquatic products(13.0%). If people did not eat seaweed foods, the results would be vegetables(47.6%), meat(19.1%) and aquatic products(18.7%), and iodine contribution rate of the 3 types of food was 85.4%. Conclusions Under the conditions of eating iodized salt, male adult dietary iodine intake is appropriate in different areas of Fujian Province. There is no such problem as iodine excessive. Iodized salt is the main source of dietary iodine. Residents in coastal city, if not eating seaweed foods, will be at risk of iodine deficiency, even if they eat iodized salt.
10.Osteopontin promotes chondrocyte proliferation in osteoarthritis
Wei JIANG ; Bowen LIN ; Xuan ZHANG ; Hua WANG ; Shuguang GAO ; Fangjie ZHANG
Chinese Journal of Tissue Engineering Research 2017;21(24):3773-3777
BACKGROUND:Osteopontin, a kind of extracellular matrix glycoprotein, has been found to participate in synthesis and catabolism of osteoarthritic chondrocyte extracellular matrix. However, the effect of osteopontin on the proliferation ability of osteoarthritic chondrocytes is little reported. OBJECTIVE:To investigate the effect of osteopontin on the chondrocyte proliferation in human knee osteoarthritis. METHODS: Cartilage samples were obtained from the patients with knee osteoarthritis undergoing knee arthroplasty at the Xiangya Hospital from January 2012 to June 2012. The chondrocytes were isolated and cultured, and then divided into four groups: blank control, osteopontin, Con-shRNA and osteopontin-shRNA groups. The cell proliferation was detected by MTT and BrdU assays. RESULTS AND CONCLUSION: After transfection of osteopontin-shRNA lentivirus, the infection rate was up to 80%. Compared with the blank control group, osteopontin group showed a significant increase in the absorbance value of osteoarthritic chondrocytes, while after osteopontin-shRNA lentivirus transfection, the absorbance value was significantly decreased (bothP < 0.05). Additionally, after osteopontin-shRNA transfection, the expression level of osteopontin was significantly downregulated (P < 0.05). To conclude, osteopontin can promote the proliferation of osteoarthritic chondrocytes, which is considered as a new treatment target for osteoarthritis.