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To establish a two-dimensional gel electrophoresis (2-DE) map for comparative proteomic analysis of rat spinal cord with chronic morphine tolerance, and to detect differentially expression proteins that are associated with chronic morphine tolerance. Methods: Sixteen male SD rats received the intrathecal catheterization operation and they were randomly divided into a morphine tolerance group (MT group, n=8) and a saline group (NS group, n=8). The lumbar enlargement segments of the MT group and the NS group spinal cord were harvested and proteins were separated by 2-DE. Differential proteome profiles were established and analyzed by means of immobilized pH gradient-based two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). The 2-DE maps were visualized after coomassie blue staining and analyzed using PDQuest analysis software. Identification of differential protein spots was conducted by MALDI-TOF-MS, and the Mascot query software was used to search Swiss-Prot database for bioinformatics analysis. Western blotting was used to verify the expression of some differentially expressed proteins. Results: A total of 1 000 spots were identified in 2-DE maps of rat spinal cord tissues from the MT group and the NS group, and 36 proteins were significantly differentially expressed in the MT group compared with the NS group. Identification was conducted by MALDI-TOF-MS and Swiss-Prot database through Mascot query software, and a total of 14 proteins were obtained. Among them, 2 protein spots were down-regulated in the MT group compared with that in the NS group, and 12 protein spots were up-regulated in the MT group compared with that in the NS group. Two kinds of proteins (NUDAA, ENOG) were verified by Western blotting and the results were consistent with proteomics data. Conclusion: The optimized 2-DE profiles for the proteome of spinal cord tissue in rats with chronic morphine tolerance is established preliminarily, which showed that morphine tolerance can cause changes in the expression of various proteins in the spinal cord.
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Animals , Male , Rats , Electrophoresis, Gel, Two-Dimensional , Morphine , Proteome , Proteomics , Rats, Sprague-Dawley , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Spinal CordABSTRACT
Objective To investigate the effect of pre-treatment of ginkgo biloba extract from portal vein on oxidative stress in liver surgery.Methods Sixty cases of hepatic portal occlusion for hepatectomy surgery in patients (38 males, 22 females, ASA grade Ⅰ or Ⅱ, cardiac function Ⅰ or Ⅱ, Child-Pugh class A), were randomly divided into three groups (n=20 each): portal vein injection group (group P), jugular vein injection group (group J) and control group (group C).Patients in group P were injected with ginkgo biloba extract injection 5 ml slowly from portal vein after freeing the portal vein, patients in group J were injected with ginkgo biloba extract injection 5 ml slowly from the jugular vein after freeing the portal vein, while patients in group C were injected with normal saline 5 ml slowly from the jugular vein after freeing the portal vein.Venous blood samples were drawn from jugular vein at the following time points: pre-occlusion ten minutes (T0), 1 h (T1), 6 h (T2), 24 h (T3) after reperfusion respectively, and then detected the levels of ALT, AST, tumor necrosis factor(TNF-α), manlondialdehyde (MDA) and the activity of superoxide dismutase (SOD).Results Compared with T0, the serum levels of ALT, AST, TNF-α and MDA at T1-T3 were significantly increased,the activity of SOD was significantly decreased in all groups (P＜0.05).Compared with group C, serum levels of ALT, AST, TNF-α and MDA at T1-T3 were significantly decreased, the activity of SOD was significantly increased in groups J and P (P＜0.05).Compared with group J, serum levels of ALT, AST, TNF-α and MDA at T1-T3 were significantly decreased, the activity of SOD was significantly increased in group P (P＜0.05).Conclusion The pre-treatment of ginkgo biloba extract from portal vein can increase the SOD activity, inhibit the activation of Kupffer cells, reduce the release of TNF-α, enhance the ability of anti-oxidative stress and produce significant protective effect on liver ischemia-reperfusion injury.
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Objective To investigate the protective effects of tanshinone-IIA sodium injection post-conditioning combined with controlled low central venous pressure on hepatic ischemia-reperfusion injury during liver resection.Methods Eighty patients scheduled for liver resection, 46 males and 34 females, aged 30-65 years, BMI 20-26 kg/m2, ASA physical status Ⅰ or Ⅱ, were randomly divided into four groups: tanshinone-IIA sodium post-conditioning (group D), tanshinone-IIAsodium post-conditioning combined with controlled low central venous pressure (CVP 1-5 cm H2O) group (group DL), controlled low central venous pressure (CVP 1-5 cm H2O) group (group L) and control group (group C) that took the static-compound anesthesia and maintained CVP 6-12 cm H2O, 20 cases in each group.The venous blood samples were drawn from internal carotid vein at different time point: pre-occlusion ten minutes (T0), post-occlusion 2 h (T1), 6 h (T2), 12 h (T3), 24 h after operation (T4), and then detected the levels of NF-κB, intercellular cell adhesion molecule-1 (ICAM-1), ALT and AST.The MAP was detected, HR and CVP were recorded.Results Compared with group C and group D, CVP were significantly lower at T0and T1in group L and group DL (P＜0.01).Compared with T0, levels of NF-κB, ICAM-1, ALT and AST in four group at T1-T4were significantly increased (P＜0.01).Compared with group C, levels of NF-κB, ICAM-1, ALT and AST in group DL, group L and group D at T1-T4 were significantly decreased (P＜0.05).Compared with group DL, levels of NF-κB, ICAM-1, ALT and AST in group D and group L at T1-T4 were significantly increased (P＜0.01).Conclusion Tanshinone-IIA sodium injection post-conditioning, combined with controlled low central venous pressure in patients with partial hepatectomy, can reduce the degree of ischemia-reperfusion injury.
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Objective To investigate the effects and mechanism of pre-treatment with an Nrf2 inducer dh404 on renal ischemia reperfusion injury in rats.Methods Thirty SD rats were divided into three groups using completely randomized digital table,dh404 was dissolved in sesame oil and was given orally 1.5 mg/kg the night before procedures and 5 hours before procedures.Rats in group Sham received no treatment of ischemic reperfusion.In group IR and group dh404,the renal ischemia reper-fusion (IR)model was established,24 hours after IR,the levels of serum creatininc (Cr)and urea ni-trogen (BUN),the activities of superoxide dismutase (SOD)and the content of malonaldehyde (MDA)in serum were measured,and hematoxylin eosin(HE)staining observe the changes in renal structure,the levels of γ-glutamate-cysteine ligase catalytic (GCLC)and modifier (GCLM)subunit, the expression of NF-κB,COX-2 and eNOS were measured.Results Compared with group Sham,the values of Cr,BUN in group IR and group dh404 were significantly higher (P <0.05).Compared to the group IR,the group dh404 Cr,BUN values significantly decreased after reperfusion for 24 h(P <0.05 ).Compared to group Sham,group IR SOD activity decreased,while the value of MDA increased(P <0.05 ).Compared to group IR,group dh404 had much higher SOD activity,while the value of MDA significantly decreased.Observed with optical microscopy,compared to group Sham, the renal tubular injury of group IR was obvious.Compared to group IR,group dh404 significantly reduced tubular injury.Compared to group IR,the levels of GCLC and modifier GCLM subunit were higher,while there were no significant differences of levels among NF-κB,COX-2 and eNOS. Conclusion Pre-treatment with an Nrf2 inducer dh404 can protect the kidney from IRI through possi-bly reducing IRI kidney oxidative stress.
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Objective To retrospectively analyze the influence of intensity-modulated radiotherapy (IMRT) on tumor regression in primary nasopharyngeal carcinoma (NPC).Methods 272 patients with NPC received radical radiotherapy alone,196 by IMRT with a total treatment time of 6 weeks,and 76 by bilateral field conventional radiotherapy (CRT) with the total treatment timc of 7 weeks.Results By the end of radiotherapy,the primary tumor and neck lymph node residual rates of the IMRT group were 36.7％ and 44.2％,respectively,both significantly higher than those of the GRT group (21.1％ and 26.6％,x2 =6.15,3.99,P ＜ 0.05).Three months after the radiotherapy,residual lesions were observed at the nasopharynx or neck lymph nodes in 12 of the IMRT group,with a residual rate of 6.1％,not significantly different from that of the CRT group (9.2％,7/76).The 12 residual lesions of the IMRT group all vanished completely 4 -9 months after the radiotherapy.Conclusions There is an obvious difference in regressive mode between IMRT and CRT technique in NPC treatment.At the end of IMRT,the tumor residual rate is slightly increased.However,the delivered dose of gross tumor volume (GTV) is sufficient,and the boost dose should not be delivered indiscreetly.
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Objective To evaluate the value of L-(methyl-11C)-labeled methionine positron emissions tomography (MET PET) and MRI in target volume delineation for postoperative radiotherapy for brain high grade glioma (HGG).Methods Thirty-seven patients with supratentorial HGG were included.Both MRI and MET PET scan were performed in the same treatment position for all patients.The consistency to determine residual tumor between MRI and MET PET was analyzed.Imaging data of MET PET and MRI were coregistered using the BrainLAB image fusion software.The extension of the volume with high uptake (VMET) on MET PET were compared quantitately with the enhancing area on MRI T1W gadolinium enhancement (VGd) and the hyperintensity area on MRI T2W (VT2).Results Both MET PET and MRI were positive for 19 patients and negative for 7 patients.The consistency between these two scans was 70.3%.MET PET was integrated with MRI in 30 patients with positive MET uptake.VMET were partially or entirely outside VGd in 29 patients and VT2 in 17 patients, whereas VGd and VT2 were partially or entirely outside VMET in all patients.The maximal distance from the margin of VMET to VGd was ≥ 2.0 cm in 50%patients and the corresponding distance of VMET to VT2 was ≥ 1.0 cm in 33% patients.Conclusions The differences are existing between MET PET and MRI in determination and identification of the location and extension of residual tumor for patients with HGG.The integration of MET PET and MRI can accurately delineate radiation target volume.