| ObjectiveBased on the experiments of immunology and molecular biochemistry this paper deals with the relativity of the level of sex hormones and the CAG polymorphisms of androgen receptor gene of the patients with acne in Hubei Province and syndrome differentiation and typing of TCM, thus finds the microcosmic theoretical basis for the TCM treatment of acne and its clinical syndrome differentiation, and the reliable evidence for the judgment of the prognosis of the patients and active therapy of TCM and Western medicine, to some extent it provides realistic theoretical basis for the application of the TCM syndrome differentiation and the disease differentiation of the therapy of TCM and Western medicine in this field.MethodsThe cases are all the acne patients in the acne specialized subject of the dermatology section in the First Hospital of Wuhan in Hubei Province. All the patients fill the case forms which are numbered. According to the syndrome differentiation of TCM we collect 93 cases of acne patients with Liver-qi Depression and 98 cases of patients with retention of damp-heat in the interior.All the testers are exsanguinated the blood 5ml from the outer veins, heparin anticoagulant, spin the tubes at 3000r/min for 5min, isolation blood serum 1.5ml into Eppendorf test tube, and put it in the refrigerator in -30℃ for testing. For the female testers we exsanguinate their blood before 7±2 days of emmenia, and avoid the emmenia and ovulation period. We adopt the radio-immunological methods to test the LH (MIU/ml), FSH (MIU/L), E2(pg/ml), T (ng/ml) and PRL (ng/ml) level of the blood serum.We select 60 random samples of the patients with Liver-qi Depression and the patients with retention of damp-heat in the interior, i.e. 30 samples of patients with Liver-qi Depression and 30 samples of patients with retention of damp-heat in the interior, then isolation the outer vein blood of 1.5ml of the above-mentioned patients, EDTA anticoagulant, and isolation the DNA of the outer vein blood leucocyte as the template.Then we select outer primers (P1: 5'- CTT TCC AGA ATC TGT TCC AGA GCG TGC-3'; P2: 5-GGCT GTG AAG GTT GCT GTT CCT CAT-3') to amplificate the microsatellite fragments of the CAG of the androgen receptor gene (reaction system of PCR is, including 25mmol/L dNTP 2ul,10×PCR Buffer 5μl,1.5mmol/L Mg2+ 3μl, 25pmol/L primer every 1μl, 5U/μl Taq DNA polymerase 0.3μl,H2O 33.5μl, template DNA 5μl;PCR program: 95℃ beforehand denaturation 5 min. Then into the cycle,94℃ denaturation 10s,65℃ annealing 30s,prolongation 60s of 74℃,cycIing 50 times), and the amplification fragments are about 202bp. 1.5% of gelose gel electrophoresis, and we get the evidence of the PCR outcome under the ultraviolet radiation lamp. We analyze it under the gel image system, and record the data with optical photography, then send the PCR outcome of the 16 random samples of thepatients with Liver-qi Depression and 14 random samples of the patients with retention of damp-heat in the interior to Shanghai Bioasis Biological Technology Co., Ltd for testing with the inside primer (5'- AGG CAC CCA GAG GCC GCG AG-3'). The result of the testing using Web Angis of Australia biological database to analyze.Results1. On the level of the serum sex hormonesThe average value (x±S) of the level of the serum sex hormones of the female patients with Liver-qi Depression:LH,3.204±2.343;FSH,2.933±2.491;E2,61.439±20.879; T,0.447±0.188;PRL, 18.388±4.568;LH/ FSH, 1.375±1.040 . The average value (X±S) of the level of the serum sex hormones of the female patients with retention of damp-heat in the interior: LH,2.927±2.786;FSH,4.817±4.581;E2,93.321±59.824;T,0.998 ±0.361;PRL,14.262±6.703;LH/FSH,0.670±0.356. Comparison between the two types: no obvious difference in LH (P>0.05); obvious difference in FSH, E2,T, PRL, LH/FSH (P<0.01); it is obvious that the average level of FSH and T of the female patients with Liver-qi Depression is higher than that of the female patients with retention of damp-heat in the interior; the average level of PRL and LH/FSH of fe... |
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