Objective To study the effects of dietary estrogens genistein(GS)and zearalenone(ZEA)on apoptosis in-duced by estrogen depletion in PEO4cells.Methods The monolayer ovarian cancer cell line PEO4cells were cultured in DMEM medium containing10%bovine serum.Before the addition of the testing compounds the cells were washed in phosphate-buffered saline and the medium was displaced with a phenol red-free DMEM medium containing5%dextral charcoal-stripped FBS and the cells were cultured for5days in order to exhaust the estrogen stored in the cells,and then cells were divided into5groups,including solvent control group,estrogen control group,anti-estrogen control group and2experimental groups.After treatment the apoptotic features of the cells were observed by cellular morphology,DNA fragmentation and location and height of cell hypodiploid were indicated by flow cytometry.Results The typical characteristics of apoptosis in PEO4cells were observed after estrogen deletion and then disappeared following exposure of the PEO4cells to32?10 -9 mol/L and96?10 -9 mol/L ZEA for72hrs.32?10 -6 mol/L and96?10 -6 mol/L GS could significantly aggravate apoptosis in PEO4cells.Conclusion Zearalenone is a kind of mycoestrogen that has estrogenic activity to inhibit apoptosis in PEO4cells.Genistein is a kind of phytoestrogen that has anti-estrogen activity(tamoxifen-like)to promote apoptosis in PEO4cells with the high doses range.

Objective To investigate the protective effect of ascorbic acid on myocardium from ischemia and reperfusion injury and its electrophysiological mechanism. Methods Using modified Ferrier's arrhythmia induced by ischemia and reperfusion injury in isolated guinea pig right ventricular wall. Results Ascorbic acid at 5, 10, 50 and 100μmol/L slightly decrease incidence of ischemia arrhythmia (P ＞ 0. 05). In contrast, ascorbic acid at 10, 50 and 100μmol/L significantly reduced the incidence of arrhythmia during early reperfusion(2～12min) (P ＜ 0. 01 or P ＜ 0. 001). Ascorbic acid at 5～ 100μmol/L didn't reverse the abbrevating of APD of endocardium and epicardium and didn't affect the conduction time in endocardium (P ＞ 0. 05). However, ascorbic acid abbreviated epicardial conduc tion time significantly (P ＜ 0. 05 or P ＜ 0. 01 or P ＜ 0. 001 ). Conclusion The study suggests that ascorbic acid might exert antiarrhythmic effect via alleviating conduction abnormity to avoid transmural reentries during ischemia and early reperfusion.

Objective To explore the effect of purification on monoclonal antibody (MAb) against PGRP by Protein A-Sepharose affinity chromatography, and to provide some based data for the purification of other antibody using the same method. Methods The ascites which include MAb was purified by Protein A-Sepharose affinity chromatography. The purity and activity of MAb was tested by SDS-PAGE and ELISA. The biological function was identified by flow cytometer and immunohistochemistry. Results The average concentration of protein in ascites before purification is 23.62 mg/ml. Before and after purification, the total protein is 148.79 mg and 146.67 mg, respectively. The recovery coefficient of protein is 98.58%. The concentration of MAb in ascites is 5.21 mg/ml averagely. The MAb purity is more than 95 %. The immunoactivity of purified antibody is higher than that of unpurified antibody. Conclusion The purity of MAb against PGRP purified by Protein A-Sepharose affinity chromatography is very high. The immunoactivity of purified antibody is higher than that of unpurified antibody. So the ProteinA-Sepharose affinity chromatography is a rapid, convenient and reliable method for the purification of MAb Against PGRP.

ObjectiveTo study the biologic distribution of 131I-Herceptin in BALB/c-neu nude mice bearing HER-2 positive SK-BR-3 human breast cancer xenografts and the radioimmunoimaging characteristics of nude mouse bearing human SK-BR-3 breast cancer xenografts. MethodSK-BR-3 breast cancer cells were implanted subcutaneously to athymic mice to establish animal model.Tumor bearing mice were continuously imaged with SPECT. The radiocounting per minute (cpm) of different organ on a γ-arithmometer was measured at 4,12,24,48 h postinjection of 131I-Herceptin or 131I-mlgG,and the T/NT ratios and the uptake percentages per gram of the injection dose (％ ID/g) was gained. ResultsModel was established in 96％ nude mouse.Compared with the control group,there was a significantly stronger contrast enhancement of tumor imaging,bigger T/NT and ％ ID/g in experimental group ( P ＜ 0.0l ).Conclusions 131I-Herceptin concentrates obviously in implanting tumor tissues of nude mouse,hence it is a good radiopharmaceutical agent targeting SK-BR-3 xenografts.

Objective To elucidate the expression and clinical significance of C-erbB-2 in the tissue of lung cancer. Methods Immunohistochemistry method was used to detect the protein expression of C-erbB-2 in lung cancer tissue. Results The positive expression rate of C-erbB-2 protein in 38 cases of lung cancer was 53.3% (21/38),which was higher than those in lung benign control group (P＜0. 001). The significant correlation were found between the protein level and tumor stage(r＝ +0. 64,P＜0.02). The order was stage Ⅳ＞stage Ⅲ ＞stage Ⅱ ＞stage Ⅰ . There was no correlation among protein expression of C-erbB-2 in various histological types of lung cancer (P＞0.05 for all). Conclusion The positive expression rates of C-erbB-2 were significantly higher in lung cancer group than those in benign control group. There is significant correlation between C-erbB-2 expression and lung cancer stage. There is no correlation among protein expression of C-erbB-2 and histological types of lung cancer.

Objective:To label monoclonal anti-GPⅢa antibody (SZ21) with 99 Tcm and evaluate the value of 99 Tcm-SZ-21 for detecting thrombi in vivo.Methods:SZ-21 was modified with 2-iminotholane and labeled with 99 Tcm-GH and radiochemical purity was determined by ITLC-SG.99Tcm-SZ-21 was injected via ear edge vein in 5 rabbits in which thrombi were induced in the right femoral arteries.As control,99 Tcm-GH was administered in 1 rabbit.Static images were acquired and irregularly shaped ROIs were drawn on the images to calculate the ratios of T/M and T/B.Vein blood was drawn at 2 min,5 min,10 min,30 min and 1 h-3 h after injection in 2 rabbits so as to observe the blood clearance of 99 Tcm-SZ-21.Rabbits were sacrificed after 3 h of imaging.The vessels including clots were harvested and imaged.Cardiac muscle,liver,spleen,lung,kidney,etc.were excised and weighed.Radioactivity counts were measured to calculate % ID/g.Results:The radiochemical purity of 99 Tcm-SZ21 was beyond 90% and stable in vitro.Thrombi could be visualized at 30 min after injection,and at 2 h image of thrombi was clearly visualized,T/M (2.55?0.72) and T/B (1.94?0.51) ratios were high.In vitro imaging showed that T/B was 4.43?1.5.Conclusion:99Tcm-SZ-21 could be a potential agent for imaging diagnosis of thrombotic disease.

0.05, respectively), but those to noradrenaline (NA: 0.01?mol/L), histamine (His: 3?mol/L), and 5-hydroxytryptamine (5-HT: 0.1?mol/L), significantly reduced (P0.05, respectively). Under atropine or Ani pretreatment, the NA-, His- and 5-HT-elicited contractions in endothelium-denuded aorta were similar to those in endothelium-intact aorta. Conclusion Atropine and Ani can inhibit receptors-mediated constrictions of rabbit aortic vascular smooth muscle cells; the actions are endothelia independent.

AIM: To investigate the electrophysiological characteristics of ventricular myocyte both in subendocardium and in subepicardium during ischemia. METHODS: Using modified Ferrier's method of ischemic and reperfusion injury in isolated guinea pig right ventricular wall. RESULTS: The transmembranal electric activities in subendocardium and in subepicardium were all significantly abnormal, and it was more significant in subendocardium. CONCLUSION: The alteration degree of ventricular myocyte in subendocardium and in subepicardium during ischemia and early reperfusion was different, and this might be the electrophysiological basis of the vulnerability of the subendocardium.

AIM: To observe the changes of interleukin-6(IL-6), IL-8 and tumor necrosis factor-?(TNF-?) in serum and lung at different time, and the effects of anisodamine (654-2) treatment in rats with oleic acid-induced ARDS. METHODS: The ARDS model induced by intravenous injection of oleic acid in the rat was used and levels of IL-6, IL-8, TNF-? in serum and lung tissue supernatant were measured using enzyme linked immunosorbent assay (ELISA). RESULTS: Levels of serum and lung tissue IL-6, IL-8, TNF-? in oleic acid type ARDS 4 h group were increased significantly. These cytokines in oleic acid type ARDS 8 h group were lower than that of ARDS 4 h group, but serum IL-6, TNF-? and lung tissue IL-6 were still higher than that of control group . In oleic acid type ARDS 16 h group, serum IL-6, TNF-? were lower than that of the ARDS 8 h group and serum TNF-? and lung tissue IL-6 were higher than that of control group. After 654-2 treatment, the levels of serum and lung tissue IL-6, TNF-? were decreased significantly. CONCLUSION: IL-6, IL-8 and TNF-? might play important roles in the oleic acid-induced ARDS in the rat. 654-2 might alleviate ARDS by inhibiting excess production of IL-6 and TNF-?.

Objective: To explore the mechanisms of the effect of isoflavone in reducing prostate cancer incidence throught studying the effects of isoflavone on apoptosis in PC-3 cell. Methods: Prostate cancer cell PC-3 was grown in RPMI 1640 medium supplemented with 10% bovine serum and 10 000 U/ml of penicillin/streptomycin in an incubator maintained at 5% CO 2 95% air and 100% humidity at 37 ℃. The respective test compound was added in fresh medium and the control cell received only the vehicle (MDSO). Apoptosis of PC-3 cells was analyzed by cell morphology under light microscope, agarose gel electrophoresis and flow cytometry. Results: Exposure of PC-3 cell to 50 ?mol/L GS, 75 ?mol/L DA and 75 ?mol/L GL after 72 h, the cell morphology indicated typical features commonly used to define apoptosis; agarose gel electrophoresis demonstrated laddered electrophoretic profiles of oligonucleosomal DNA fragments indicative of apoptosis, and flow cytometric analysis revealed a hyperdiploid population in the tested cells. Conclusion: Isoflavones could induce apoptosis in prostate cancer cells PC-3 and it may be the main cause of their role in cancer inhibition.