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Objective To construct and identify adenovirag (Ad-RUNEP) carrying human B-endorphin(B-EP) genes which can be regulated by mifepfistone (RU486)-inducible system and to evaluate the effects of different concentrations of RU486 on the transgene expression in adenovirus in vitro.Methods The shuttle plagmid pDC312一RUNEP carrying B-EP genes which can be regulated by RU486-inducible system was constructed and was combined with adenovims to form a recombinant Ad-RUNEP using AdMAXTM system.The recombinant Ad.RUNEP was then amplified and purified.The titers of the adenovirus were determined and the adenovirus vector was checked.After being infected by Ad-RUNEP for 24 h,A431 cell line was incubated in liquid culture media containing RU4860,10-10,10-9,10-8,10-7,and 10-6 mol/L respectively(group R0-5) for 48h,and Was then transferred to liquid culture media containing no RU486.The liquid culture media were obtained on 1st.2nd and 4th day of incubation and centrifuged.The supematant Was collected for determination of B-EP concentration by ELISA.Results The analysis of enzyme-incision demonstrated that RU486 regulating system and B-EP were cloned directly into pDC312-RUNEP.The titer of Ad-RUNEP was 2.25×1010 pfuml.The expression of B-EP was significantly higher in group R1-s than in group R0 and was significantly lower in group R1-3 than in group R4 (P<O.05).The expression of B-EP wag significantly higher on 2nd than on 1st and 4th day.Conclusion The adenovims carrying B-EP genes which can be regulated by RU486 (Ad-RUNEP) Wag successfully constructed.
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Objective To compare the effects of volume preloading with crystalloid and colloid fluid to prevent hypotension associated with thoracic epidural block combined general anesthesia.Methods Ninety ASA Ⅰ～Ⅱ grade patients,scheduled for elective upper abdominal surgery,were randomly allocated to three groups(30 patients in each group).A,B and C group respectively received 1000ml lactated Ringer's solution,500ml and 1000ml polygeline injection for volume preloading in 40min before general anesthesia induction.During volume preloading,epidural catheter was placed at T_ 8～9 and blocked with mixed solution of 1.6% lidocaine and 0.2% dicaine.Results Systolic blood pressure(SBP) and diastolic blood pressure(DBP) were significantly lower in group A and B than those in group C after epidural block,general anesthesia induction and intubation(P
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0. 5mv. LAD occlusion was maintained for 50min and then released for reperfusion (120min) . Myocardial infarct size was measured by nitroblue tetrazolam at the end of experiment. Results There was no statistically significant difference in HR, MAP, cardiac output (CO), left ventricular systolic pressure (LVSP), left ventricular end-diastolic pressure ( LVEDP) and rate-pressure products ( RPP) among all groups during ischemia and early reperfusion period except for LVEDP in control group which was much higher than that in all preconditioning groups during the late reperfusion period. Compared with the control group, desflurane, sevoflurane, isoflurane and G+ desflurane reduced myocardial infarct size by 41%, 47%,31.7% and 17.8% respectively without significant homodynamic effects. Myocardial infarct size in G + desflurane groups was significantly larger than that in desflurane group. Conclusion Preconditioning with desflurane, sevoflurane and isoflurane reduce myocardial infarct size in rabbits to some extent. The protective effects may be partly medialed via activation of KATP channel.
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The plasma level of endogenous digitalis-like substance(EDLS) was measured in 15 patients who underwent cardiac operation under cardiopulmonary bypass (CPB). Blood samples were taken before and after anesthesia induction,immediately before CPB and removing the aortic crossclamp,and at 5,30min after heart rebeating. Resluts showed that the plasma EDLS level slightely decreased after anethesia indction, and significantly decreased immediately before removing the aortic crossclamps and at 5 min after heart rebeating (P
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Objective To assess the changes in the effect compartment concentration (Ce) and bispectral index (BIS) during a step-by-step TCI of propofol in the elderly.Methods Ten ASA Ⅰ -Ⅱ patients (6 male, 4 female) aged 67-77yr and weighing 51-78kg, undergoing elective surgery were studied. Patients with severe cardiovascular disease were excluded. The patients were unpremedicated. Anesthesia was induced with propofol administered by a TCI system (Diprifusor) . The target concentration (Ct) of propofol was increased step-by-step from 1?g?ml-1 to 4?g?ml-1 in 6 steps. At each step Ct increased by 0.5?g?ml-1 and the interval between the two steps was 2min. The changes in Ce (calculated and displayed by Diprifusor) and BIS were recorded. Modified OAA/S sedation score were measured. Blood samples were taken from artery for determination of blood concentration of propofol (Cb) before TCI (T0 ) and when Ct was 1, 2, 3, and 4?g*******ml-1 (T1-4) in 5 randomly selected patients.Results (1) Ce consistently increased with the increase in Ct and there was a delay between Ct and Ce. When Ct was increased to 4?g*******ml-1, it took (14.4 ? 0.5) min to achieve the balance between Ct and Ce. (2) Cb of propofol was higher than Ct of propofol at each step. MDPE and MDAPE was 9.7% and 11.2% respectively. (3) There was a close correlation between BIS and Ct, Ce and OAA/S score( r = - 0.878, - 0.888 and 0.913 respectively , P
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Objective To evaluate the roles of K ATP channels in desflurane-induced myocardial protection from anoxia/reoxygenation injury Methods Primary cultured rat myocardial cells were randomly allocated to four groups: control group(A): without any treatment; anoxia/reoxygenation group(B): reoxygenation of 1 h following anoxia of 2 h; desflurane preconditioning group(C): 20 min of 9% desflurane preconditioning followed by 10 min washout before anoxia/reoxygenation and K ATP channel blocker group(D): adding glybenclamide at final concentration of 12?g/ml to culture medium 10 min before the same procedures as group C The activities of lactic dehydrogenase (LDH) and creatine kinase (CK), rates of cell viability and apoptosis, contents of cellular malondiadehyde (MDA) and adenosine triphosphate (ATP), and intracellular free calcium concentration were measured Results Compared with control group, anoxia/reoxygenation caused great increases of levels of LDH, CK, apoptosis and MDA ,and decreases of ATP and cell viability (P
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0 05); the increased degree of only AST activity reduced in PE group(P
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6 h (n = 20). Swan-Ganz catheter was left in place after surgry for 40h. MAP, HR, ECG, cardiac output (CO), cardiac index (CI), PAP, PCWP, CVP, SVR, PVR and DO_2,VO_2, O_2-extraction rate (O_2 ER) were measured and/or calculated immediately after operation (T_1), 16 h (T_2 ),24 h (T_3 ) and 40 h (T_4 ) after operation. Results VAS scores were significantly lower in PCIA group than thosein IM group at T_(2, 3, 4) (P