| Objective:The peri-menopausal period is the female transitional stage from the period of duration to the senium. The physiological change of this period represented by the gradually dysfunction of the ovary, the ovarian atrophy and the obviously decreased of the estradiol excretion. The broken balance of the hypothalamic-pituitary-ovarian axis resulted in a series of symptoms such as hectic fever, nervousness, depression, irritability, insomnia and so on, namely the Climacteric Syndrome. With the change of the living conditions, the contagious deseases had been better controlled, however, the incidence rate of the Climacteric Syndrome had the increasing tendency. More and more people attached great importance to the living qualities of the aged people suffered from this . In recent years, the studies to the mechanism of the Climacteric Syndrome mainly focused on the field of generendocrine and neuroendocrine. Although the studies explained the mechanism of the Climacteric Syndrome in some degree, there were still other questions we couldn' t answer. So it was necessary to study the mechanism of the syndrome in another field.It was proved that Chinese medicine therapy had great effect on the Climacteric Syndrome. And its little side effct was also acceptable.Our previous clinical researchs and experiments of the Gengniananyi tablet (GNAT) showed that the GNAT had good effects on the syndrome and the actovoty ratio reached 96% ;and the GNAY had the effcts of adjusting the nerve function, regulating the generation endocrine, enhancing the immunological competence, preventing and curing the bone rarefaction and the cardiovascular diseases, postponeing the ageing, and it was safe and had no poison.This research would filter the significative genes about cell signal transduction through the change of the suffer' s genes between they hadn' t been cured and had been cured, and from the change of transcription level of the gene about cell signal transduction to discuss it with gene chip. From which we would make preliminary exploring on the molecular level about the mechanism of GNAT curing the syndrome.Methods:This research regarded the patient of the deficiency of the kidney and spleen type as the object. The patient was treated by GNAY for 4 weeks. We getted 5ml blood from peripheral vein of the patient before the teatment and anticoagulate it with Heparin, quickly separated the leucocytes and added lml Trizol agent, blended, scattered the test tube untill the mixture looked clear, bright and not viscous, then put it into the refrigerator at 4°C. After that we drew the RNA in the cell with the one-step and condensed RNA through isopropyl alcohol deposition, purified the total RNA, measuredit with spectrophotometer at last. Then they were measured in quality by lectrophoresis with sagauoe. Crossing and rinsing, the labeled RNAs were dissolved in the crossed liquid and were stayed over with crossing at 42 °C. After being crossed, they were rinsed in the liquid at 42°C, then cleaned in 0. 2XSSC at room temperature. Been swinged and dried, the slides were scaned with dual channel laser scanner. Dealed with the blood of the patient after the treatment in the same way. The test had used two chip, one is labeled with Cy5 as before the treatment and Cy3 as after. Another was labeled with Cy3 as before the treatment and Cy5 as after, in order to avoid the difference of absorbency. Then all the number of the foreground subtracted the backgroud, and drew the fluorescence signal number of Cy5> Cy3. Collecting the picture of the chip and analyzing the date : analyzing the picture, we turned the picture signal into number signal, grouped all the data together with Lowess'method, then confirmed the difference of the expressed gene at the standard by double difference.Results:1. The results of RNA purified: absorbency of OD26O/28O>1. 9, 28S and 18S clear- visible, 5S was loom, 3 banding were not degrade. It explained that the total RNA was intact.2. The crossing pseudo color picture of the model chip , Cy5 exhibited by red and Cy3 exhi... |
PDF Full Text Request