| PrefaceShortage of insulin secretion absolutely or relatively and the reduction of target cell sensitivity to insulin, diabetes mellitus can result in metabolism disorder of sugar, protein, fat and water-electrolyte. At the present time, diabetes has developed very fast and become the 5th death reason all of the world. Type 2 diabetes is the major type of diabetes that brings increase of the incidence. With reforming and opening in our country, development of economic and the change of people s living form, the incidence of diabetes is rising day by day and becomes a serious public health problem.Normally, people think that type 1 diabetes is an autoimmune disease which has organ specificity, it has surely hereditary foundation, triggered by various environmental factors and mediated by T cell. The cause of type 2 diabetes hasnt been understood very much yet, in most cases, people think this disease perhaps relates with the heredity factor because it has obvious hereditary trend and heterogeneity. With the research of many immune cells during these years, im-munopathological reactions mediated by T cell and the functions of other cytokine factors play the important roles in diabetes development. The test is to study the change of T cell subsets in obese and non-obese patients who are suffered from type 2 diabetes. Now, reportis as follows.MaterialSpecimen source: All blood samples of experiment groups are taken from the endocrine ward of china medical university affiliated the first hospital. According to the diagnosis standard option of ADA, 46 persons who have been suffered from type 2 diabetes are chose. According to the difference of body mass index ( BMI) , we divided them into two groups. Testees whose BMI > 27 are set up for the obese group; 19s, 12 male examples, 7 female examples, 30 -64 years old, 47. 84 ±8. 54 years old in average. Course of disease 15 days -15 years, 5.02 ±4.33 years in average. Testees whose BMI <23 are set up for the non-obese group: 27s, 14 male examples, 13 female examples, 39 ~ 73 years old, 57. 48 ± 10. 09 years old in average. Course of disease 10 days - 20 years, 8.21 ±6. 15 years in average. None of all has the immune system diseases and urgently chronic influence history. The normal control group is selected from the healthy blood donors in the hospital, aged 19-46 years old. The crowd which BMI > 27 are set up for obese control, 7 male examples, 5 female examples. The crowd whose BMI <23 are set up for non-obese control, 8 male examples, 5 female examples. Picking blood method; Take venous blood 3ml, put it in the tube with heparin, split T lymphocytes and mark with monoclonal antibody. The sample must be examined immediately to avoid the death of WBC.Test instrument and reagent; flowcytometer, centrifuge and oscillator etc. Major reagents are PE-CD3 monoclonal antibody, FITC-CD4 monoclonal antibody, FITC-CDg monoclonal antibody (purchased fromFrench Immunotech company) and lymphocytes Separation Medium.MethodProcess; Transfer centrifugal blood on lymphocytes Separation Medium slowly, centrifuge at 2500rpm 10min, separate the lymphocytes , get 3 test-tubes and mark them with 1, 2 and 3. Add CD3 (10μl)and CD4(10μl) antibody in the first test-tube, CD3(10μl) and CD8( 10μl) antibody in the second test-tube, no antibody in the third test-tube. Vibrate 20 seconds to make well mixed. Place the mixture in 4± for 30 -45minutes, wash with PBS and centrifuge at 3000rpm x 5min, repeat this procedure three times, send the sample into the flowcytometer and record the results.ResultThe T cell subsets of type-2 diabetes obese patient measure results as follows: CD3 + , range 48. 15 ~ 87.46, 70. 10 \ 10. 67 in average ; CD4 + , range 21. 00 ~ 69.00, 38.49 \ 11. 25 in average; CDg + , range 11.47 -33.77, 23. 08 \4. 93 in average, CD4/CDg, range 0. 86 -4. 86, 1. 77 \0.84 in average. The T cell subsets of type 2 diabetes non-obese patient measure results as follows; CD3 + , range 46. 92 - 78.66, 61.46 \ 9.52 in average; CD4 +, range 22.18 - 54...
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