| Immune system do not existence and perform it's function Solitary, but contact tight with nervous system and endocrine system, on some characteristic cells which regulate internal environment by immune mechanisms, there are also molecul receptors having regulation function to nervous system and endocrine system, so there is neuroim-munology and neuroendocrinology concept in modern. When nervous system and endocrine system are interfered with by environment, it is inevitable to effect the function of Immune system, especially the Specificity Immune system.During perioperation, the interference with Immune system from tension, anxious, drugs, wounds, anesthesia, etc, had caused our attention in the increasing of infection ratio and tumor repea. At present, most studies focus on the function of different anesthesia to relieving immunosuppression from stress of operation, but it has few reports about the effects of epidural block on Immune system in the condition of without stress of operation.ObjectiveTo observe the effects of epidural block with ropivacaine on T -lymphocyte subsets ?natural - killer cell and immunoglobulin -IgA,IgM in the condition of without stress of operation.Material and Methods45 patients requiring epiclural anesthesia for undering bladder or lower extremity surgery were randomly divided into three groups; Group H was given high concentration local anesthetic (1.0% ropiva-caine) ; Group L was given low concentration local anesthetic (0. 5% ropivacaine ) ; Group N - the control group was given only testing dose and identical Saline. At 30min before entering operation room, mid-azolam 50ug/kg and atropine 10ug/kg were given to all patients of three groups. Monitoring ECG,HR,BP,SPO2 and set up vein -via. Puncture at left - lateral position, change to supine position after puncturing and placing catheter. If there were no symptoms of injecting in blood vessel and entire spinal cord narcosis 3min after giving test dose 1% lidocaine, specified concentration of ropivacaine or saline 20ml were given according the difference of groups by 5min interval, Stabing technique was used to determine block range every 3min until stabilizing of pain sense blocking range, the range up to T8 -T10 and down to S3 - S5. 10 - 15mg efedrina was given to the patients occurrencing Severe hypo tension and 0. 3 -0. 5mg atropine was given to the patients occurrencing bradycardia. Before giving test dose ( pre - block .T1 ) , 30min,60min,90min ( T2 ,T3 ,T4) after stabilizing of pain sense blocking range (normal control group based on the average value of force - experiment: 13.5 ±4. 8min) , two samples of peri -venous blood were obtained; one was 1.5ml, anticoagulated with EDTA; The other was 4ml, centrifugated at 3000r/min for 5min, then 2ml serum was obtained, putted in - 80℃ cryogenic refrigerator stored and waited for measurement.Measurement of T - lymphocyte subsets - CD3+ , CD4+ , CD8+, ( CD3+ + CD4+ )/( CD3+ + CD8+ ) , natural - killer cell ( CD16++56) : Place 20ul two -color direct immunofluorescence reagent (Simultest IMK - Lymphocyte) in each sample tube , use a fresh micropipetter tip and carefully add 100ul anticoagulated whole blood into the bottom of the labeled tubes, mix and incubate for 30min, then add 1 x Lysing Solution into each tube, mix and incubate for 12min (do not exceed 12min) in the dark, centrifuge tubes at 2500r/min for 10min after washing with CellWASH, add 0.5ml fixing solution (0.1% paraform-aldehyde) ,mix and then place in the dark , be ready to be analyzed on the flow cytometer.Measurement of immunoglobulin: ( 1) establish standard curve: dissolve working standard labeled with the content of IgG,IgA,IgM with 0. 5ml distilled water, then dilute it with saline to 5 standard solution with different concentration. Add standard solution 10ul for IgG,IgA and 20ul for IgM into 5 antero - hole of each monophasic im-munodiffusion plate, place in level moist box and incubate in 37℃ he-mo iothermy incubator, observe results 48h later, measure cyclic s diameter of diffu...
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