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BACKGROUND: Rabbit serum samples are widely used in basic researches, and two-dimensional gelelectrophoresis (2-DE) is the most classic technique for protein separation. Therefore, it is of great significance to establish a stable technique system of 2-DE for rabbit serum.OBJECTIVE: To establish a 2-DE technique system for rabbit serum protein separation. DESIGN, TIME AND SETTING: A single sample observational experiment was performed at Tianjin Key Laboratory of Biomarkers for Occupational and Environmental Hazard of Chinese People's Armed Police Forces Medical College from June to July in 2008. MATERIALS: Six healthy rabbits were provided by the Animal Experimental Center of Tangshan Vocational Technical College METHODS: Health rabbit serum was dissolved in rehydration sample loading buffer before and after eliminating high abundance proteins to make proteins in it schizolysis adequately. After reductive alkylation, the samples were loaded into the rehydration tray to undergo passive rehydration for 14 hours. Isoelectric focusing (IEF) electrophoresis was followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). After sliver staining, the gel was analyzed by PDQuest7.4. MAIN OUTCOME MEASURES: ①Efficiency of eliminating high abundance proteins. ②Two-dimensional eleotrophoregrams (2-D electrophoregrams)RESULTS: Distinct 2-D eleotrophoregrems were obtained with high resolution and good reproducibility. The removal of high abundance proteins in serum failed to result in better 2-D electrophoregrams.CONCLUSION: We have successfully established a 2-DE technique for rabbit serum proteome, which can lay the foundation for the further study of serum proteomics of diseases.
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Objective To observe the effects of 6%hydroxyethyl starch(Volven 130/0.4)infusion on hemodynamics and coagulation.Methods 60 cases of open heart surgical children under cardiopulmonary bypass were randomly divided into two groups(30 cases for each):The control group(group C)were infused with 400 ml plasma and volven group(group V)were infused with 400 ml Volven before cardiopulmonary bypass.Then mean aterial pressrue(MAP),heart rate(HR),and central venous pressure(CVP)were measured.The venous blood samples were collected before and after infusion for the measurements of the following parameters:haematocrit(HCT),blood sedimentation(BSR),platelet count(PLC),prothrombin time(PT),and activated whole blood clot time(ACT).Those indexes before operation,15 min after bypass,after returning to ICU,and 24 hours after operation were observed.Results HCT and PLC after infusion were significantly lower than that before infusion.BSR after infusion was significantly faster than that before infusion.There were no significant changes in MAP,HR,CVP,PLC,ACT and PT.Conclusion 6%Hydroxyethyl starch infusion is safe and beneficial during cardiopulmonary bypass.
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BACKGROUND: Limb ischemia reperfusion (LIR) as a stressor leads to gastric mucosal injury, and then results in the occurrence of stress ulcer.OBJECTIVE: To observe the effects of LIR on gastric mucosal injury, investigate part of the mechanism, and the role of several transient limb ischemia in the occurrence of gastric mucosal injury.DESIGN: A randomized grouping design and controlled animal experiment.SETTING: Department of Pathophysiology of North China Coal Medical College.MATERIALS: The experiment was carried out in the pathophysiological laboratory of North China Coal Medical College from January to June 2002. Fifty-four healthy adult male Wistar rats were randomly divided into three groups with 18 rats in each group. Ischemia reperfusion (I/R) group: The rats were duplicated into models according to the Rosenthal method that under superficial anesthesia with ether, the roots of both hindlimbs were ligated by wrapping with rubber strap, blood flow was blocked for 4 hours and then recovered to perfusion for 4 hours, and finally killed by bleeding from abdominal aorta. Ischemic preconditioning group: Before model establishment, blood flow of both hindlimbs was blocked for 5 minutes, and then recovered to perfusion for 5 minutes, which was repeated for four times, and the following operations were the same as those in the I/R group. Control group: The operations were the same as those in the I/R group,but both hindlimbs were ligated at relaxation without blocking the blood flow.METHODS: Sections of gastric mucosa were prepared, and then observed under light microscope and electron microscope, and the index of gastric mucosal injury was determined according to the Guth standard. The colorimetric assay was performed with 721 spectrophotometer at 650 nm, and the amount of gastric combining mucus was calculated.Meanwhile, the blood flow of gastric mucosa, contents of phospholipid and hexosamine in gastric mucus, content of nitric oxide in plasma and gastric tissue and activity of nitric oxide synthase (NOS) in gastric mucosa were determined.MAIN OUTCOME MEASURES: Index of gastric mucosal injury, amount of gastric combining mucus, blood flow of gastric mucosa, contents of phospholipid and hexosamine in gastric mucus, contents of nitric oxide in plasma and gastric tissue and NOS activity in gastric mucosa.RESULTS: All the 54 rats were involved in the analysis of results. ① In the I/R group, gastric mucosal injury was serious, edema, hyperemia, erosion and disintegration of gland of mucosal glands were observed, infiltration of inflammatory cells (formation of ulcer) was observed in basal and inferior mucosa. In the ischemic preconditioning group, the gastric mucosa was complete, and the damaged severity was milder than that in the I/R group; Under electron microscope, the organell structures of gastric parietal and chief cells were incomplete and destroyed. The cell injuries in the ischemic preconditioning group were milder than those in the I/R group (index of injury: 18.00±10.71, 34.00±15.01, P＜ 0.01). ② The blood flow and combining mucosal amount of gastric mucosa, contents of phospholipid and hexosamine in gastric mucus in the ischemic preconditioning group and I/R group were all obviously lower than those in the control group [(2.12±0.56), (10.84±2.56), (25.52±2.97) mL/(kg·h); (2.01±0.91), (2.79±0.73), (3.99±0.87) mg;(7.68±1.95), (9.74±1.04), (11.98±1.98) mg/g; (3.83±1.18), (5.42±0.47), (5.76±1.21) mg/g, P ＜ 0.05, 0.01], those the above indexes were all higher in the ischemic preconditioning group than in the I/R group. ③ The contents of nitric oxide in plasma and gastric tissue and NOS activity in gastric mucosa in the ischemic preconditioning group and I/R group were significantly lower than those in the control group [(250.0±5.6), (270.0±11.3), (210.0±7.4) μmol/L; (9.34±0.67), (11.34±1.00), (7.50±0.67) μ kat/g, P ＜ 0.01], those were also signficantly higher in the ischemic preconditioning group than in the I/R group.CONCLUSION: As a stressor, LIR can lead to gastric mucosal injury, and cause stress ulcer.Ischemic preconditioning can alleviate the gastric mucosal injury following LIR
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AIM: To observe the degree of gastric mucosal injury following limb ischemia/reperfusion(LIR),and to investigate the mechanism of gastric mucosal injury and the influence of the series of brief ischemia/reperfusion of hind limbs of rats on gastric mucosal injury. METHODS: Referring to Rosenthal's method, the model rats which underwent 4 hours of ischemia and 4 hours of reperfusion in hind limbs were made. The indexes of gastric mucosal injury after LIR and ischemic preconditioning (IPC) + LIR were determined. The morphologic changes were observed with light microscope and transmission electron microscop respectively. The GMBF,histologic lesion score, gastric barrier mucus, phospholipids, hexosamine, nitric oxide and nitric oxide synthase in mucus were measured in different groups. RESULTS: Serious damage in gastric mucosa was observed under microscope and EM after LIR. But less serious damage was observed in the IPC group. After LIR, compared to the control group, the GMBF and the content of gastric barrier mucus, phospholipids and hexosamine in mucus decreased significantly. There was significant difference in most indexes between the control group and the IPC group, but compared to LIR group, the histologic lesion score decreased significantly and the GMBF and the content of gastric barrier mucus, phospholipids, hexosamine ,nitric oxide and nitric oxide synthase in mucus increased significantly. CONCLUSION: LIR caused the gastric mucosa injury. IPC alleviated the damage of gastric mucosa following ischemia/reperfusion in hind limbs.
