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Phospholipase Cγ1 and NF-κB are required for cell-matrix adhesion of colorectal cancer cells

Xiumei LI ; Xiaochun BAI ; Fan DENG ; Di LU ; Shenqiu LUO

Academic Journal of Second Military Medical University.2005;26(5):465-470.

Objective: To investigate the function and mechanism of phospholipase Cγ1 (PLCγ1) in cell-matrix adhesion in colorectal cancer. Methods: Highly metastatic colorectal cancer cell line LoVo and lowly metastatic colorectal cancer cell line SW480 were subjected to cell-matrix adhesion assay. U73122 (a specific inhibitor of PLC) and pyrrolidine dithiocarbamate (PDTC) (an inhibitor of NF-κB) were used to study the effect of PLCγ1 and NF-κB on cell-matrix adhesion. Furthermore, Western blot and gel electrophoresis mobility shift assay (EMSA) were performed to detect the mechanism of PLCγ1 in colorectal cancer cell adhesion to matrix. Results: Inhibition of PLCγ1 or NF-κB resulted in reduction of cell-matrix adhesion in a dose-dependent manner in LoVo cells(P<0.05), but had no marked effect on SW480 cells. Western blot analysis showed that epidermal growth factor (EGF) stimulated the phosphorylation of PLCγ1 in LoVo. The results of EMSA indicated that inhibition of PLCγ1 signaling pathway also down-regulated the activity of NF-κB while EGF reversed the function. Conclusion:These data suggest that PLCγ1 plays a pivotal role in the EGF-induced cell-matrix adhesion of highly metastatic colorectal cancer cells and that NF-κB is also functional in this signaling pathway.

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Expression, purification and characterization of PCP-2EC/Fc fusion protein in mammal cells

Peng ZHANG ; Hexin YAN ; Hongyang WANG

Academic Journal of Second Military Medical University.2005;26(3):300-304.

Objective: To construct the extracellular region of PCP-2(PCP-2EC) and the immunoglobin IgG Fc fusi on protein expression vector,and then express and purify the soluble PCP-2EC/Fc fusion protein for the study of its function in neuronal adhesion. Methods: PCP-2 extracellular region was amplified and cloned into an expression vector pIGplus containing human IgG Fc; PCP-2EC/Fc fusion protein was expressed by COS-7 and 293 cells transfected by the constructed plasmid and purified by protein A. The purified fusion protein was used as substrate to study its function in neuronal adhesion. Results: PCP2 extracellular region was cloned into IgG Fc expression vector successfully; PCP 2EC/Fc fusion protein was expressed and purified in mammal cells; and the purified fusion protein promoted neuronal adhesion. Conclusion:PCP 2EC/Fc fusion protein expression system is successfully constructed and the purified fusion protein can promote neuronal adhesion. These results lay a foundation for the research on the PCP-2 function in neuronal adhesion and the further functional study in the nervous system and other fields.

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Study on epithelial-mesenchymal transition in the early stage of mice renal interstitial fibrosis

Jun XIONG ; Kaimeng HU ; Kaihong JI ; Shuping TANG ; Yin WANG ; Houqi LIU

Academic Journal of Second Military Medical University.2005;26(4):355-360.

Objective: To observe morphological changes of epithelial-mesenchymal transition in the early stage of mice renal interstitial fibrosis. Methods: Renal interstitial fibrosis was induced by unilateral ureteral obstruction(UUO) in mice. Histological and immunohistochemical methods were used to analyze pathological changes and α-SMA expression in renal tissue.Argentum hexamethylenamine staining and transmission electron microscopy were used to observe changes of the renal tubule basement membrane. Gelatin zymographic analysis was used to observe the expression of MMP2 and MMP9 in renal tissue.Results:The mice suffered from renal interstitial fibrosis were identified by histological analysis and α-SMA positive cells in renal tissue. Argentum hexamethylenamine staining and transmission electron-microscopy showed that the renal tubule basement membrane disrupted locally and renal tubule epithelial cells invaded into the renal interstitium in the early stage of renal interstitial fibrosis. Gelatin zymographic analysis showed that the expression of MMP2 and MMP9 was increased transitorily in the early stage of renal interstitial fibrosis. Conclusion: Renal tubule basement membrane disruption, renal tubule epithelial cells invasion into the renal interstitium, and the expression of MMP2 and MMP9 are involved in the development of renal interstitial fibrosis.

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Heme oxygenase-2 gene deletion protects basal ganglia cells from oxidative injury induced by free Fe2+

Yan QU ; Xiang ZHANG ; Jianning ZHANG ; Haining ZHEN ; Dakuan GAO

Academic Journal of Second Military Medical University.2005;26(4):390-395.

Objective:To determine whether heme oxygenase-2(HO-2) gene deletion can attenuate oxidative injury induced by free Fe2+. Methods:Stereotactic injection of 10 μl sterile FeCl2 (10 mmol/L) was made into the right striata of HO-2 knockout mice and wild-type mice. Brain edema severity was measured at 24 h. Cell viability, protein oxidation, and lipid oxidation of the basal ganglia were determined at 72 h. Western blot analysis was applied for heme oxygenase-1 (HO-1) measurement.Results: Brain water content significantly decreased in HO-2 knockout mice at 24 h compared with wild-type mice. Protein oxidation and lipid oxidation significantly decreased in HO-2 knockout mice at 72 h compared with wild-type mice, while the striatal cell viability increased significantly. HO-1 expression at baseline and 72 h was also similar to that in wild-type mice. Conclusion:These results show that HO-2 gene deletion can protect basal ganalia cells from free Fe2+ -mediated oxidative stress injury,suggesting that selective inhibition of HO-2 may have a protective effect on brain oxidative injury.

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Effects of soybean isoflavone on liver oxidative stress resulting from 60Co-gamma rays

Lihua SONG ; Donglian CAI ; Hongli YAN ; Xiaoli CHEN ; Yahui LI ; Li MA ; Tongjie HU

Academic Journal of Second Military Medical University.2005;26(2):151-154.

To study the effect of soybean isoflavone on liver oxidative stress resulting from 60Co-gamma rays. Methods: Totally 80 normal female Kunming mice were evenly randomized into 5 groups according to body weight: 3 intervention groups, single irradiation group and normal control group. The normal group and single irradiation groups were given 0.5% CMC-Na, and the 3 intervention groups were given different doses of soybean isoflavone (50 mg/kg, 100 mg/kg, 400 mg/kg) respectively for 14 d. The whole body of single irradiation group and intervention groups were subjected to 4.56 Gy 60Co-γ radiation once on the 7th day, and then the mice were killed on the 2nd day and the 7th day after radiation. Results: The CAT activity of liver tissue of 100,400 mg/kg intervention groups and 3 SI groups were significantly increased on the 2nd day and 7th day after irradiation(P<0.05), respectively; the GSH-Px activity of 100 mg/kg SI group was significantly increased(P<0.05) on the 7th day after irradiation; the T-SOD activity of 50 mg/kg SI group was significantly decreased (P<0.05) on the 2nd day after irradiation,while no difference was observed among remaining groups. The MDA content of 100 mg/kg group was significantly decreased on the 7th day after radiation compared with control group, and MDA content of each group subjected to irradiation were increased on the 2nd day after irradiation,but 3 SI groups nearly decreased to normal level on the 7th day after irradiation. Conclusion: The soybean isoflavone can enhance the antioxidant capability of mice, but it does not show a dose-effect relationship.

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Electrophysiologic study of f-wave amplitude in chronic atrial fibrillation associated with rheumatic heart disease

Li LI ; Baocheng JIA ; Baoren ZHANG ; Zengwei WANG ; Jialin ZHU

Academic Journal of Second Military Medical University.2005;26(2):127-130.

To investigate the electrophysiologic characteristics of f-wave amplitude and to evaluate its role in development and persistence of chronic atrial fibrillation (AF) associated with rheumatic heart disease (RHD). Methods: Epicardial mapping was performed in 44 patients with chronic AF of RHD who underwent heart valve surgery. Ten patients with supraventricular tachycardia served as the control group. Results:The f-wave amplitude of left atrium (LA) and middle and low LA posterior regions were significantly lower than those of the control group. The f-wave amplitudes of the upper, middle and low sections in LA posterior region were significantly lower than those in right atrium (RA) (P<0.05). The f-wave amplitudes were compared before and after electrocardioversion in 14 patients with chronic AF. The mean atrial electrogram amplitude during sinus rhythm was significantly higher than that during AF (P<0.01).The f-wave amplitude in left appendage was higher than that in LA posterior region (the upper,middle and the lower part),P<0.05.The f-wave amplitude in the upper section of LA was significantly higher than that in the middle section. The f-wave amplitude in AF group was not correlated to the diameter or volume of both atria. Conclusion: There are amplitudes differences between the upper, middle and lower LA,suggesting that the middle and lower sections of LA posterior wall may be the region producing anisotropy and reentrant circle.

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Clinical application of transbronchial needle aspiration

J.f.turner JR ; Kopen WANG

Academic Journal of Second Military Medical University.2004;25(7):746-750.

Transbronchial needle aspiration(TBNA) is able to provide definitive answers for a wide variety of pulmonary benign and malignant diseases including infectious diseases,cystic lesions, granulomatous diseases, pulmonary and mediastinal mass, etc. It's proved particularly revolutionary to stage bronchogenic carcinoma during routine flexible bronchoscopy. What's more, TBNA is an extremely safe procedure with no reported major complications. This review introduces the techniques about identifying the target, obtaining the specimen and preparing and interpreting the specimen when conducting TBNA. The complications, a through unusual, and major limitations are also touched in this review.

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Arterial baroreflex function and left ventricular hypertrophy

Chaoyu MIAO ; Dingfeng SU

Academic Journal of Second Military Medical University.2004;25(4):448-448.

It is well known that the arterial baroreflex(ABR)plays a key role in the regulation of heart rate and stabilization of blood pressure.Currently,it appears that ABR dysfunction is involved in the pathophysiology of cardiovascular disease states.Since the mid-1990s,a number of studies have been carried out in our laboratory to explore the pathological significance of ABR function in cardiovascular damage.This minireview summarizes our research work on the topic of ABR and left ventricular hypertrophy(LVH).On the basis of discussion concerning the importance of ABR dysfunction in hypertensive LVH and sinoaortic denervation-induced LVH,we advance a new strategy for reversal of LVH,that is,restoration of impaired ABR function.We tested this hypothesis in animal models with ABR deficiency.It was found that improvement of impaird ABR function with long-term treatment of ketanserin or candesartan was accompanied by reversal of LVH.The preliminary results indicate that it is feasible to target ABR for treatment of LVH.

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Effects of glucose-insulin-potassium on baroreflex sensitivity, left ventricular function and ventricular arrhythmia in the subacute phase of myocardial infarction in rats

Jianguo LIU ; He SHU ; Fuming SHEN ; Chaoyu MIAO ; Dingfeng SU

Academic Journal of Second Military Medical University.2004;25(4):433-433.

Objective:Glucose-insulin-potassium(GIK) is clinically used for reducing mortality in acute myocardial infarction(MI). It is known that ventricular arrhythmia, left ventricular dysfunction and impaired baroreflex sensitivity(BRS) are the three major determinants for predicting the mortality after acute MI. The present work was designed to study the effects of GIK on BRS, ventricular arrhythmia, and left ventricular function in rats with coronary artery ligature. Sprague-Dawley rats were used and the myocardial infarction was produced by ligature of the left anterior descending artery. Five weeks after coronary artery ligation, BRS was measured in conscious state with a computerized blood pressure monitoring system and left ventricular function and electrocardiogram were determined in the anaesthetized state in the subacute phase of myocardial infarction. It was found that GIK did not affect the blood pressure and heart period in both conscious and anaesthetized rats. GIK did not enhance BRS, but reduced ventricular arrhythmia and improved left ventricular function by reducing left ventricular end diastolic pressure in anaesthetized rats with MI. It is proposed that reducing ventricular arrhythmia and improving left ventricular function contribute to the effect of GIK on reducing the mortality after MI.

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Contribution of blood pressure variability to the effect of nitrendipine on end-organ damage in spontaneously hypertensive rats

Jianguo LIU ; Liping XU ; Zhengxu CHU ; Chaoyu MIAO ; Dingfeng SU

Academic Journal of Second Military Medical University.2004;25(4):406-406.

Objective:It has been proposed that blood pressure variability(BPV) is positively related to end-organ damage(EOD) in hypertension.The present work was designed to observe the effects of long-term treatment with nitrendipine and hydralazine on BPV and EOD in spontaneously hypertensive rats(SHR),to examine the hypothesis that lowering BPV with an antihypertensive drug is an important factor in organ protection.Design and methods:Drugs were mixed in rat chow.After 4 months of drug administration,blood pressure was recorded continuously in conscious freely moving rats for 24 h.The heart,kidneys,and brain were then isolated and examined.Results:It was found that nitrendipine significantly decreased blood pressure and BPV,and significantly decreased EOD score in SHR.Hydralazine decreased blood pressure,but did not lower BPV.No effect on EOD was found in hydralazine-treated rats.In control rat(n=38),EOD score was weakly related to systolic blood pressure(r=0.331,P<0.05) and closely related to long-term systolic BPV(r=0.551,P<0.01).In nitrendipine-treated rats,EOD score was closely related to long-term systolic BPV(r=0.602,P<0.01),but not to blood pressure level(r=0.174,P>0.05).Conclusion:BPV plays an important role in the organ-protecting effects of nitrendipine.

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