0.05) and group TP2 ( P

Objective To construct the recombinant adeno-associated virus vector with human preproenkephalin gene (rAAV-hPPE). Methods The human preproenkephalin (hPPE) gene was cloned into the adeno-associated virus (AAV) vector plasmid pSNAV which contained neo expression box. The recombinant pSNAV-hPPE was then transfected into BHK cells using lipofectamine?2000. The G418-resistant cells, BHK / SH1, were obtained. The BHK / SH1 cells were infected with HSV1-rc /△UL2 which has the function of packaging the recombinant AAV(rAAV) . After purification, the construction of rAAV-hPPE was achieved.Results The construction of pSNAV-hPPE was confirmed by digestion with restriction enzyme. Southeon bolting was used to detect the virus liters (2.5 ? 1212 v.g /ml) .Conclusion This rAAV-hPPE virus vector with high liter and strong infectivity can be used in transgenic analgesic research.

Objective To investigate the effect of ketamine on spatial learning and memory in a rat model of acute pain produced by an incision in plantar area. Methods Seventy-two 3-month old male SD rats weighing 200-250g were randomly divided into 3 groups: (1) control group (C) received no plantar incision (n = 24); (2) acute pain group (M) received an incision in the sole of the hindpaw according to the method described by Brennan (n = 24) and (3) ketamine group (K) received plantar incision and intraperitoneal ketamine 10 mg?kg-1 every day for 7 days starting from the day when plantar incision was made ( n = 24). In group C and M normal saline (NS) was given i.p. instead of ketamine. The three groups were further divided into 2 subgroups according to the time when Morris water maze (MWM) testing was started-1 week (C1, M1,K1) or 3 weeks (C3, M3, K3) after plantar incision. MWM test was performed 4 times a day for 6 consecutive days. The latent periods and swimming distances were recorded automatically by MWM monitoring system. Six days after the last MWM testing the animals were anesthetized and killed. The hippocampus was removed for microscopic examination. Results (1) MWM test: From the 1st to the 6th day of testing, the latent period and swimming distance in group K1 were significantly longer than those in group C1 and M1 (P

Objective To investigate the protective effects of isovolemic hemodilution with 6% hetastarch (HAES) and ligustrazine on myocardium against ischemia/reperfusion injury. Methods Thirty-two healthy male rabbits were anesthetized with 3% pentobarbital 30 mg?kg-1 , tracheotomized and mechanically ventilated. Left femoral artery and vein were cannulated for direct BP monitoring, blood collection and fluid infusion. Chest was opened and myocardial ischemia was produced by temporary ligation of the anterior descending branch of left coronary artery. Myocardial ischemia was confirmed by elevation or depression of S-T segment and /or high T-wave. The animals were randomly divided into 4 equal groups with 8 animals in each group: Ⅰ control group was subjected to 45 min myocardial ischemia followed by 180 min reperfusion without any treatment; Ⅱ hemodilution group in which 9 ml Ⅲ kg-1 blood was removed and blood volume was maintained by simultaneous infusion of equal volume of 6% HAES at 20 min after myocardial ischemia was started; Ⅳ ligustrazine group received ligustrazine injectio 20 mg?kg-1 iv 20 min before and 40 min after myocardial ischemia was started; Ⅱ hemodilution + ligustrazine group in which hemodilution was performed as in group Ⅲ and ligustrazine injectio was given iv as in group Ⅲ . BP and HR were recorded before during and at the end of myocardial ischemia, and at 30, 60, 120, 180 min of reperfusion. Hct was measured before and after hemodilution. Blood samples were taken for determination of plasma CPK and LDH activities before ischemia (T0), at the end of 45 min ischemia (T1) and at the end of 180 min reperfusion (T2 ) . At the end of the experiment, myocardial tissue 0.2 g was obtained from ischemic and non-ischemic area for determination of myocardial CPK and LDH activities and election microscopic examination. Results (1) In all four groups plasma CPK and LDH activities were significantly increased after ischemia (T1 ) and were increased further after reperfusion (T2) (P

Objective To determine the mimic epitope of N-methyl-D-aspartate receptor 2B subunit (NR2B). Methods The monoclonal antibody interacting with NR2B was used as target protein to screen the binding peptide from a 12-mer Ph. D random peptide library. After three rounds of affinity screening, twelve specific clones were selected randomly and identified by sandwich ELISA. The peptide sequences were analyzed by DNA sequencing. The clones containing a common sequence were named positive clone. The competitive inhibition of the native antigen bound to monoclonal antibody against the NR2B by the positive clone was assayed by cell ELISA. Results After 3 rounds of screening the phages specifically bound with mAb NR2B were selected and amplified. Nine of the 12 clones displayed a common aminoacid sequence: SHPPVMPWPTST. The inhibitory assay showed that the mimic epitope peptides displayed on the phage surface could effectively inhibit the combination of the monoclonal antibody with native antigen. The inhibitory rate of mimic epitope was (45?3)% .Conclusion The mimic epitope of NR2B was screened successfully from the 12-mer Ph D random peptide library. This peptide mimics an epitope of the native NR2B.

Objective To evaluate the imaging characteristics of the bone infarct . Methods 13 cases of the bone infarct proved by clinical manifestation and surgical pathology were collected,there were 5 males and 8 females. All patients had X-ray films, 4 were imagedwith CT scanning , 7 were imaged with MRI , and 2 underwent all three techniques . The bone infarct were classified into the early,themiddle and later stages.X-ray , CT and MRI findings of bone infarct ,were analysed characteristics . Results Partial osteoporosin and punctate calcifications could be seen on X-ray and CT , in the early and middle stages , which presented as iso-/low-signal on T_1WIand iso-/high-signal on T_2WI in the center of the bone infarct with a tortuous low-signal on T_1WI and high-signal on T_2WI zonearound.In the later stages,the bone infarct was characterize by irregular sclerosis and tortuous calcification on X-ray and CT,and by low-signal on both T_1WI and T_2WI . Conclusion MRI is the most effective way to find the bone infarct in the early stages , X-ray is helpful in diagnosing the bone infarct in the middle and later stages , CT is more sensitive than X-ray . X-ray ,CT and MRI should be combined in diagnosing the bone infarct .

Objective: To perform target-controlled infusion (TCI) in total intravenous anesthesia (TIVA) of propofol and fentanyl. Method: Using effect compartment modeling, computer controlled infusion(the computer software was developed by Coezee and Pina) was performed during induction and maintenance of anesthesia in two groups of adult patients. The target predicted concentration of theoretical effect-site compartment for propofol was 4?g/ml and for fentanyl was 2?g/ml. The plasma concentration (Cm) of propofol was determined by fluorospectrophotometry and Cm of fentanyl was measured with radioimmunoassay. Result: The mean Cm from 0 to 120 min showed that excessive dose of propofol was administered, MDAPE=25%,however the mean Cm of fentanyl was lower than the target level obviously,MDAPE=35.5% in first group. After an imitative calculation,another pharmacokinetic (PK) parameter sets of propofol and fentanyl were selected in the second group,MDAPE=15.5% for propofol and MDAPE=37. 75% for fentanyl. Conclusion: The concentrations of propofol and fentanyl in the effect site compartment can be achieved rapidly by using the effect compartment control algorithm. The PK parameter,described by different authors influences the accuracy of TCI administration.

0.05). Conclusion:The stress reaction to human injury stimulation are inhibited effectively by propofol - fentanyl intraveous anesthesia

Objective To study the effects of profound hemodilution with 6% HES (200/0.5) on coagulation.Methods Ten male new Zealand long-ear rabbits weighing (2.43 ?0.19) kg were anesthetized with intravenous thiopentone 8 mg-1 and tracheotomized and mechanically ventilated (VT = 15 ml-1 , RR = 24 bpm) . Anesthesia was maintained with intravenous infusion of thiopentone and succinylcholine. Femoral artery and vein were cannulated for BP and CVP monitoring. Hemodilution was performed in 6 steps at 30 min intervals. Blood was withdrawn from artery and simultaneously replaced by intravenous infusion of equal volume of 6% HES until Hct was 5 % -8 % . Blood samples were taken before hemodilution and 30 min after each step of hemodilution for determination of coagulation function curve using Sonoclot coagulation and platelet function analyzer. Results When Hct was 25%-30% there was no significant difference in coagulation function before and after hemodilution. When Hct was 15%-20% there was significant difference in ACT, TTP, clot rate and MCS before and after hemodilution but the normal coagulation process was not affected. When Hct

Objective To investigate the effects of nuclear factor κB(NF-κB) antisense oligodeoxynucleotides (AS-ODNs) on transforming growth factor β1 (TGF-β1)-induced epithelial mesenchymal transition (EMT) in human renal tubular epithelial cells. Methods NF-κB AS-ODNs were transferred into the human renal tubular epithelial cells (HK-2), and the cells were stimulated by 10 μg/L TGF-β1 for 24 hours. The expression of NF-κB mRNA and α-SMA mRNA were measured by RT-PCR. α-SMA protein expression was assessed by fluorescence spectrum.Results TGF-β1 significantly up-regulated the expression of NF-κB mRNA, which was 8 folds of blank control (P<0.01). TGF-β1-indueed epithelial mesenchymal transition was inhibited by NF-kB AS-ODN and the NF-KB mRNA expression of AS-ODNs was decreased by 75%(P<0.05).The expression of α-SMA mRNA and protein was also down-regulated obviously (P<0.05).Conclusion NF-κB AS-ODN can inhibit the expression of NF-κB and the epithelial-mesenchymal transition, which may be a new therapeutic strategy against tubulointerstitial fibrosis.