| Preparation of Adsorbent for Whole Blood Immunoadsorption of MG and Observation of it's BiocompatibilityObjective To prepare the desired adsorbent for immunoadsorption of anti-AChR antibodies in MG patients and observe the biocompatibility of this adsorbent. Methods The carrier were prepared, i.e. cellulose beads activated by epichlorohydrin, L-tryptophan chosed as the ligand. The ligand was immobilized on the carrier to make adsorbent. The healthy rabbits were divided into two groups, i.e. control group (without adsorbent) and experimental group (with adsorbent). The rabbits underwent extracorporeal whole blood adsorption for 120 minutes and blood samples were withdrawn before and after hemoperfusion. Blood cells were counted, plasma electrolytes, blood lipids and proteins were assayed. Student t test was used for statistic analysis.Results The adsorbent was prepared with cellulose as carrier and tryptophan as the ligand. Control group showed no significant changes of blood cells, plasma protein, blood lipids and electrolytes in the extracorporeal blood circuit. Blood cells, blood lipids and electrolytes were not damaged or altered by this adsorbent, but the concentration of the plasma protein dropped down. Total protein of experimental group decreased from 56.9±4.56 to 49.7±6.22 (g/l, P<0.05), and globulin protein decreased from 28.7±3.86 to 22.4±6.38 (g/l, P<0.05), but albumin protein doesn't altered.Conclusion The adsorbent with cellulose as carrier tryptophan as ligand was biocompatible with the blood system. Extracorporeal whole blood circuit was safe enough for the blood cell, plasma electrolytes, blood lipids and proteins. Theexperiment itself couldn't influence the analysis of the results.Passive Transfer of Experimental Autoimmune Myasthenia Gravis in Rabbit with Human Anti-Acetylcholine Receptor AntibodiesObjective To induce passively transferred experimental autoimmune myasthenia gravis (EAMG) rabbits for the study of therapy on myasthenia gravis (MG)and observe clinical manifestation, electrophysiological function and the light microscopic alterations in EAMG rabbits.Methods Sera obtained from the myasthenia gravis patients and normal people were stored at -70 °C. Crude immunoglobulin fractions were prepared from pooled sera by ammonium sulphate precipitation at 33% saturation. Crude immunoglobulin was purified by chromatography on DEAE-Sephadex A-50. The 24 healthy rabbits were divided into three groups, i.e. experimental group (receiving IgG of myasthenia gravis patients), healthy control group (receiving IgG of normal pepole) and control group (not receiving IgG). The IgG was injected intraperitoneally(i.p) in one dose for 3 days. Clinical manifestation was graded in 4 levels. Electrophysiological function was assessed by repetitive nerve stimulation (RNS), single fibre electromyography (SFEMG) and miniture endplate potential(MEPP) tests. Anti-acetylcholine receptor antibodies were evaluated by enzyme linked immunosorbent assay (ELISA). Histological studie were evaluated by immunohistology. Student t test was used to analyze the difference among the three groups.Results The second day after the passive injection, all the rabbits of experimental group showed clinical signs of fatigability and weakness. The weakness exacerbated on the third day. Clinical symptoms could be improved by neostigmine. Anti-acetylcholine receptor antibodies in experimental group was significantly higher than that of the healthy control group and control group (P<0.01). Thepositive ratio of anti-acetylcholine receptor antibody in experimental group was 8/8, and that in the healthy control group and control group was 0/8. At 3, 5 > lOHz, the decrement of compound muscle action potential (CMAP) and the mean jitter (MCD) of all the rabbits of experimental group were significantly higher than the healthy control group and control group(PO.Ol). The percentage of decrement of CMAP in the order of control group, healthy control group and experimental group were:3Hz: 1.000±0.756,l .25±0.463,21.875±3.220,5Hz:l .250±0.707,l .500±0.925,22. 250±2.815,10Hz:1.125±0.641,1.625±0.744,24.375±1.685. Following the above sequence, the MCD (}j.s)were 3Hz:6.375±2.503,6.75±2.314,23.125±2.997,5Hz:7.625±3.159,8.125±2.997,26.375±4.172,10Hz:9.625 ±1.922,10.125±1.807,30.375±4.470. The amplitude and frequency of MEPP in experimental group decreased significantly comparing with that of healthy control group and control group(P<0.01).The amplitude(uV) in the order of control group, healthy control group and experimental group were 49.33±4.86,9.07±4.67,30.54±3.74. Following the above sequence, the frequency(/sec) werel9.45±2.21,19.11±2.48,17.18±2.87. The quantity of neuromuscular junction per unit area(25mm2) in the order of control group, healthy control group and experimental group were 12.81±2.56,12.4±2.29,9.83±3.40. The quantity of neuromuscular junction per unit area(25mm2) in experimental group was decreased significantly comparing with the healthy control group and control group(P<0.05). Conclusion Passive transfer the IgG of myasthenia gravis patients to rabbits could successfully induce the EAMG rabbits, accompanied by clinical signs of weakness,elevation of anti-acetylcholine receptor antibodies, blockage of neuromuscular transmission and decrement of the neuromuscular junctions. Neostigamine could improve the manifestation of the rabbits.
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