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Objective To evaluate the role of mitochondrial permeability transition pore (mPTP)in reduction of brain injury by sevoflurane postconditioning in a rat model of hemorrhagic shock and resuscitation (HSR).Methods Ninety pathogen-free healthy adult male Sprague-Dawley rats,weighing 300-350 g,were divided into 5 groups (n =18 each) using a random number table:sham operation group (group S),group HSR,sevoflurane postconditioning group (group SP),sevoflurane postconditioning plus atractyloside (ATR,a specific mPTP opener) group (group SP + ATR) and ATR group.Hemorrhagic shock was produced by withdrawing 40％ of the total blood volume from the right carotid artery over an interval of 30 min,and 1 h later the animals were resuscitated by infusion of the shed blood via the left jugular vein over 30 min.SP and SP+ATR groups were exposed to 2.4％ sevoflurane for 30 min starting from the onset of reinfusion.In ATR and SP+ATR groups,ATR 5 mg/kg was intravenously injected at 10 min before reinfusion.Six rats in each group were randomly sacrificed at 24 h after the end of autologous blood reinfusion,and the hippocampus was harvested for determination of the expression of Bcl-2 and Bax in hippocampal tissues (by Western blot) and degree of mPTP opening.At 72 h after the end of autologous blood reinfusion,the rest 6 rats in each group were selected and underwent Morris water maze test,and the cognitive function was evaluated.Results Compared with group S,the escape latency was significantly prolonged,the number of crossing the original platform and locomotor distance in the target quadrant were decreased,the expression of Bcl-2 was down-regulated,the expression of Bax was up-regulated,and the degree of mPTP opening was increased in group HSR (P＜0.05).Compared with group HSR,the escape latency was significantly shortened,the number of crossing the original platform and locomotor distance in the target quadrant were increased,the expression of Bcl-2 was up-regulated,the expression of Bax was down-regulated,and the degree of mPTP opening was decreased in group SP (P＜0.05),and no significant change was found in each parameter in ATR and SP+ATR groups (P＞0.05).Compared with group SP,the escape latency was significantly prolonged,the number of crossing the original platform and locomotor distance in the target quadrant were decreased,the expression of Bcl-2 was down-regulated,the expression of Bax was up-regulated,and the degree of mPTP opening was increased in group SP+ATR (P＜0.05).Conclusion The mechanism by which sevoflurane postconditioning ameliorates brain injury may be related to inhibiting mPTP opening in a rat model of HSR.
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Objective To evalute the role of phosphatidylinositol 3?kinase(PI3K)∕serine?threo?nine kinase(Akt)∕endothelial nitric oxide synthase(eNOS)signaling pathway in sevoflurane postcondi?tioning?induced attenuation of brain injury in a rat model of hemorrhagic shock and resuscitation(HSR). Methods Seventy?two pathogen?free healthy adult male Sprague?Dawleg rats, weighing 300-350 g, were divided into 4 groups(n=18 each)using a random number table: sham operation group(group S), group HSR, sevoflurane postconditioning group(group SP)and sevoflurane postconditioning plus PI3K∕Akt signaling pathway specific inhibitor wortmannin group(group SP+WT). Hemorrhagic shock was in?duced by withdrawing blood(40% of the total blood volume)from the right common carotid artery over an interval of 30 min, and 1 h later the animals were resuscitated with infusion of the shed blood via the left jugular vein over 30 min. In group SP+WT, wortmannin 0.6 mg∕kg was administrated via the jugular vein at 30 min before establishment of the model. In SP and SP+WT groups, 2.4% sevoflurane was inhaled for 30 min starting from the onset of infusion of the shed blood. At 10 min before withdrawing blood(T0), im?mediately after the end of withdrawing blood(T1), at 30 min and 1 h after the end of withdrawing blood (T2,3)and immediately after infusion of the shed blood(T4), blood samples from the common carotid ar?tery were collected for blood gas analysis, the blood lactate concentration was recorded, and mean arterial pressure was simultaneously recorded. At 24 h after infusion of the shed blood, 6 rats were randomly select?ed from each group and sacrificed, and their brains were immediately removed for determination of cerebral infarct volume(by TTC staining), expression of hippocampal caspase?3(by immuno?histochemistry), and expression of Akt, phosphorylated Akt(p?Akt)and eNOS(by Western blot). The ratio of p?Akt∕Akt was calculated. Results Compared with group S, the mean arterial pressure was significantly decreased and the blood lactate concentration was increased at T1?3, the cerebral infarct volume was increased, and the expression of caspase?3 was up?regulated in the other three groups, and the ratio of p?Akt∕Akt was sig?nificantly increased, and eNOS expression was up?regulated in group SP(P<0.05). Compared with group HSR, the cerebral infarct volume was significantly decreased, the expression of caspase?3 was down?regula?ted, the ratio of p?Akt∕Akt was increased, and eNOS expression was up?regulated in group SP(P<0.05). Compared with group SP, the cerebral infarct volume was significantly increased, the expression of caspase?3 was up?regulated, the ratio of p?Akt∕Akt was decreased, and eNOS expression was down?regula?ted in group SP+WT(P<0.05). Conclusion PI3K∕Akt∕eNOS signaling pathway activation mediates sevoflurane postconditioning?induced attenuation of brain injury in a rat model of HSR.
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Objective To investigate the inhibition effects of an hTERT-promoter-dependent oncolytic adenovirus Ad-VT that expresses apoptin on human prostatic carcinoma cell PC-3. Methods MTT assay was used to measure viability of PC-3 cell which was infected by recombinant adenovirus.The viability was measured at time points of 12,24,36,48,60,72,84 and 96 h after infection.AO/EB staining,DAPI staining,Annexin V assay were used to investigate the lethal effect and style of Ad-VT on PC-3 cell in vitro.The Caspases were measured by whole cell extraction of PC-3 cells 48hrs after infection. Results Ad-VT,Ad-VP3 and Ad-GT inhibited the proliferation of PC-3 cell in vitro.Ad-VT and Ad-GT were more effective than Ad-VP3 on cell growth,P ＜ 0.05.At 48,72,96 h time points,the inhibition effect of Ad-VT on PC-3 cell exhibited a dose related manner.When infection at MOI 100,the inhibition effect of Ad-VT on PC-3 cells exhibited time related manner.The AO/EB staining,DAPI staining,Annexin V assay,Annexin V assays and Caspase assays showed that Ad-VT inhibited the proliferation of PC-3 cells by inducing apoptosis of prostate cancer cells,Loss of cytoplasmic membrane integrity. Conclusions The hTERT-promoterdependent oncolytic adenovirus Ad-VT could effectively suppress prostate cancer cells PC-3 growth.
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Objective To analyze the risk factors of fevers after percutaneous nephrostolithotomy (PCNL) and to determine a more effective prophylaxis method.Methods A retrospective analysis of 320cases who underwent PCNL for renal calculi from 2008 to 2011 (men 233,women 87,age between 22 years to 72 years) was made.The average age of the patients and the average diameter of the stones were 42 years and 3cm(0.8 -6 cm),respectively.We analyzed factors such as age ( ＞60 years and ≤60 years),stone size ( ＞2.0 cm and ≤2.0 cm),operative time ( ＞60 min and ≤60 min),irrigation pump pressure ( ＞120 mm Hg and ≤ 120 mm Hg),obstructive conditions,preoperative urinary tract infection and fever.Wethen compared the fever rate with each of the risk factors.Results There were 59 cases with fever after PCNL; 18.4％ of the cases had a temperature over 38 ℃.There were two cases of pyemia.The patients whose stone diameter ＞ 2 cm had a post-operative fever rate of 22.4％.The fever rate in patients whose stone diameter ≤2.0 cm was 10.4％.The fever rate in patients with an operative time ＞60 min and ≤60 min was 27.2％ and 10.4％,respectively.Patients with irrigation pump pressure ＞ 120 mm Hg and ≤ 120mm Hg had post-operative fever rates of 28.3％ and 11.0％,respectively.There was statistical significance between each post-operative fever risk factor group.Conclusions The post PCNL fever risk factors are stone diameter ＞ 2.0 cm,operation time ≤60 min and irrigation pump pressure ＞ 120 mm Hg.The effective prophylaxis policy of post-operative fever are the pre-operative using of broad-spectrum antibacterial agents,shorter operative time and lower irrigation pump pressure.A two-stage operation procedure will also reduce the post-operative fever.
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Objective To discuss the cause and management of delayed hemorrhage after minimally invasive percutaneous nephrolithotomy (PCNL). Methods From 2004 to 2009,there were 13 cases (10 men and 3 women, mean age 47 years) developed severe bleeding following PCNL. The cause of hemorrhage and treatments were retrospectively analyzed and summarized. Results The time of hemorrhage was 5-40 d. In 7 of 13 cases, the bleeding were was controlled by complete bed rest,hemostatic,balloon compression and clamped nephrostomy tube. Six of 13 underwent selective renal arteriography after ineffective with conservative treatment. The DSA showed 4 were false aneurysm and 2 were arteriovenous fistula. All the 6 cases were treated by the super-selective arteriolar embolization. The hematuria was disappeared 1- 3 d later. The intravenous pyelogram revealed the renal function kept well during follow-up visit. Conclusions Delayed hemorrhage is one of the severe complications, which may be caused when the renal puncture passage established as forming false aneurysm and arteriovenous fistula. The renal arteriography and super-selective arteriolar embolization could be a safe and effective treatment for the severe hemorrhage after PCNL.
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Objective To investigate the efficacy of Holmium laser for upper urinary tract diseases.Methods A total of 101 patients with urinary tract stricture or calculus in the lower or middle ureter were treated with holmium laser by ureteroscopy from January 2005 to June 2007 in our hospital.For the patients with stones in the upper urinary tract or the kidney,percutaneous nephrolithotripsy(PNL) with holmium laser by ureteroscopy was performed.Results No patients developed perforation,urinary leakage,or infection after the operation.In the 23 cases of ureteral stricture,6 were cured,10 were improved,5 were failed,and 2 were deteriorated or even developed hydronephrosis.The two who were deteriorated after the operation was converted to open surgery because of pain in the kidney area and refractory infection.They were cured after resection of the strictured segment and end-to-end anastomosis.Follow-up was available for 3 to 24 months(mean,12 months);none of the patients had recurrence during the period.In the 40 patients with lower or middle ureteral stones,nephrolithotripsy was completed successfully in all but 7,in whom the stones moved into the kidney during the procedure.These 7 patients were then cured by extracorporeal shock wave lithotripsy.The 40 patients were followed up for 3 to 18 months(mean,15),none of them had recurrence.In the 24 cases of upper urinary tract stones and 14 patients with renal calculus,the stones were removed completely by the first PNL in 36;2 patients received a second operation after indwelling renal cannula for 1 week because of intraoperative hemorrhage.These 38 patients achieved a follow-up of 3 to 24 months(mean,18),no one of them showed residual stones,or recurrent calculus or stricture of the ureter.Conclusions Holmium laser by ureteroscopy is safe,effective,and minimally invasive for patients with upper urinary tract disorders.The treatment is worth being used thanks to a low rate of intra-and postoperative complications.