OBJECTIVETo evaluate the effects of Carvedilol on cardiopulmonary bypass (CPB)-induced myocardiocyte apoptosis and its effects on regulation of Fas, FasL expression, caspase-3 activity and oxidative stress in the left ventricle (LV) in this setting.

METHODSTen adult dogs undergoing conventional hypothermic CPB were randomly divided into control and Carvedilol treated groups (n = 5, respectively). Dogs in Carvedilol treated group received a bolus of Carvedilol (1 mg/kg) intravenously and a maintenance dosage of Carvedilol (3 micro g.min(-1).kg(-1)) for 3 hours after the reperfusion of the heart. Dogs in control group received no Carvediolol. LV samples were obtained before, during and 3 hours after CPB. In situ nick end-labeling (TUNEL) technique was used to detect the apoptotic cells. The expressions of Fas and FasL were detected immunohistochemically and quantified by fluorescence activated cell sorting (FACS). The activity of caspase-3 enzyme and malondialdehyde (MDA) level were measured by cleavage of Z-DEVD-AMC substrate and thiobarbituric acid reactive substance (TBARS) method, respectively.

RESULTSBefore and during CPB, all the parameters were not significantly different intra- or between groups (P > 0.05). After CPB, these parameters in both groups were significantly elevated compared with those of before and during CPB (P < 0.028, respectively). However, the number of apoptotic cells in Carvedilol treated group was significantly decreased compared with that of the control group (P < 0.021). The expressions of Fas and FasL were significantly downregulated by Carvedilol (P < 0.001 and 0.003, respectively). The caspase-3 activity and the content of MDA in the Carvedilol treated group was also significantly reduced (P < 0.026 and 0.005, respectively).

CONCLUSIONSCarvedilol significantly reduces CPB-induced cardiomyocyte apoptosis in dog hearts and the reduction of cardiomyocyte apoptosis is associated with downregulation of Fas and FasL expression, inhibition of caspase-3 activity and oxidative stress in LV.

Adrenergic beta-Antagonists ; pharmacology ; Animals ; Apoptosis ; Carbazoles ; pharmacology ; Cardiopulmonary Bypass ; Caspase 3 ; Caspases ; metabolism ; Dogs ; Down-Regulation ; Fas Ligand Protein ; Female ; In Situ Nick-End Labeling ; Male ; Membrane Glycoproteins ; metabolism ; Myocytes, Cardiac ; cytology ; metabolism ; Propanolamines ; pharmacology ; Signal Transduction ; fas Receptor ; metabolism

Objective To compare epididymis-sparing orchiectomy (group A) with traditional orchiectomy (group B ) in patients with advanced prostate cancer,and to evaluate which procedure is better.Methods A total of 60 cases of advanced prostate cancer patients were enrolled,with 30 cases in group A and 30 cases in group B.They were given oral anti-androgen from 1 day after castration.Serum level of testosterone and prostatic specific antigen (PSA) was detected before castration,and 1 week,1,3,6,9 and 12 months after castration.Patient satisfaction was also evaluated.Results On time point of 12 months after castration,the average level of serum testosterone was 0.2 nmol/L (95 ％ confidence interval,0.1 ～ 0.9 nmol/L) in group A and 0.3 nmol/L (95％ confidence interval,0.2 ～ 0.9 nmol/L) in group B (P ＞0.05 ) ; the average value of PSA was 0.22 ng/ml in group A and 0.27 ng/ml in group B (P ＞0.05 ) ; patient satisfaction rate was 96.7％ (29/30) in group A and 53.3％ (16/30) in group B.Conclusions No significant difference of testosterone level and PSA is found between the 2 groups.However,epididymis-sparing orchiectomy meets the psychological needs better because it helps to maintain the appearance of the scrotum through epididymis preservation and epididymoplasty.

Whether conventional hypothermic CPB induces myocyte apoptosis in dog hearts and modulation of bcl-2, bcl-xl, bax, bad, and caspase-3 pathways in this setting was investigated. Ten healthy adult dogs were randomized into sham-operated and CPB groups. Samples of left ventricle were obtained before, during and 3 h after CPB. In situ TUNEL was used to detect apoptotic myocytes. Immunohistochemistry and flow cytometry were employed for detection of expressions of bcl-2, bcl-xl, bax and bad proteins. Z-DEVD-AMC substrate cleavage and TBARS methods were used to measure the activity of caspase-3 and the content of lipid peroxide in LV myocardium, respectively. After CPB, the number of apoptotic myocytes in CPB group was significantly increased. The results of immunohistochemistry demonstrated that bcl-2, bcl-xl, bax and bad proteins were constitutionally present on the sarcolemma of the LV myocytes. FACS results showed that, after CPB, expressions of bax and bad in CPB group were significantly upregulated, while the expressions of bcl-2 and bcl-xl were not significantly changed in both groups. The activity of caspase-3 and the content of lipid peroxide in LV myocardium in CPB group were also significantly increased after CPB. The present study shows that there exists myocardiocyte apoptosis in dog hearts undergoing conventional hypothermic CPB and the myocyte apoptosis is initiated by ischemia and performed during reperfusion. Moreover, the CPB-induced myocyte apoptosis was associated with upregulation of expressions of bax and bad proteins, activation of caspase-3 and increase of oxidative stress.
Animals ; Apoptosis ; Cardiopulmonary Bypass ; Caspases ; metabolism ; Dogs ; Female ; Hypothermia, Induced ; Lipid Peroxidation ; Male ; Myocytes, Cardiac ; metabolism ; pathology ; Proto-Oncogene Proteins ; genetics ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; Random Allocation ; bcl-2-Associated X Protein

OBJECTIVE To heterologously express and purify bioscavenger organophosphorus hydrolase(OPH),and evaluate its ability to resist ethyl paraoxon poisoning in vivo.METHODS The Escherichia coli expression strain of OPH was constructed and purified by Ni-column affinity chroma-tography and gel filtration chromatography,and the purified product was identified by mass spectrometry.The enzyme activity and kinetic constants(Km,Vmax,kcat and kcat/Km)were measured using ethyl paraoxon as the substrate.Twelve SD rats were randomly divided into the experimental group and control group.The experimental group was given 1 mg·kg-1 of OPH solution by iv administration while the control group was given the same volume of normal saline.After administration,the two groups were immedi-ately sc administration with 2×LD50 ethyl paraoxon(0.86 mg·kg-1).The state of the rats was observed and the poisoning symptoms were scored.The survival rats were given 2×LD50 ethyl paraoxon every 24 h,and the survival curve and symptom score chart were drawn according to survival and poisoning symp-toms of the rats to evaluate the anti-organophosphorus poisoning ability of OPH.RESULTS The expres-sion strain of OPH in E.coli was successfully constructed.After two-step purification,a single band of OPH was obtained by sodium dodecyl sulphate-polyacrylamide gel electrophoresis,indicating that OPH had high purity,and the prepared protein sequence was consistent with the target protein.For OPH,Km=7.5×10-5 mol·L-1,Vmax=2.2×10-7 mol·L-1·s-1,kcat=158.4 s-1,kcat/Km=2.1×106 L·mol-1·s-1.The rats in the control group showed obvious poisoning symptoms after being given 2×LD50 ethyl paraoxon,and all rats died with in 15 min.The rats in the experimental group did not show poisoning symptoms after the first exposure,and the poisoning symptoms gradually deepened after continuous exposure until all the rats died on the 4th day.CONCLUSION OPH with high purity is successfully prepared in this study,and OPH could effectively resist ethyl paraoxon poisoning in rats.