The Expression And Significance Of Vascular Endothelial Growth Factor And Basic Fibroblast Growth Factor In Endometrium Bleeding Induced By Intrauterine Device

Objective: With the development and consummation of IUD , the acceptance of the child-bearing period women was risen and the application range was broaden . But the common ill-effect was still abnormal uterus bleeding , such as menorrhagia,menostaxis , spotting bleeding anterior-mensturation and post-menstruation . The mechanism was multiple factors , dependended on endometrium vascular's generation and plerosis . The vascular's gereation was regulated by vascularization factor . At present , VEGF and bFGF were two promoting vascularization factors followed with interest , although their mechanisms of promoting vascularization were different , they were all regulated by estrogen and progestogen , they coordinated and cooperated during the process of endometrial blood vessel generation and plerosis . The objection of this study was utilizing the hysteroscope technique , drawing the endometrium under the location by looking steadily , adoption immunohistochemical (IHC)method and computer image analysis technique . To study the expression and significance of VEGF and bFGF in endometrium bleeding induced by Intrauterine Device (IUD) , to find the new idea of IUD bleeding and provide the theory basement for clinical therapy from the vascularization mechanism. Method : The patients were selected from the outpatients of Obstetrical and Gynecology, second Affiliated Hospital, Hebei Medical University .Ⅰgroup : 30 patients were abnormal uterus bleeding after inserting IUD , 15 patients with metal "O" ring and 15 patients with "T" ring . Ⅱgroup : 16 patients were normal menstrual cycle after inserting IUD , 8 patients with metal "O" ring and 8 patients with "T" ring . Ⅲ group: 15 patients were normal menstrual cycle without IUD . Each group was comparable on the age , the time of inserting ring and past history. The patients of normal mensterual cycle were selected to check on the 3-7 days after menstruation clean. The patients of abnormal bleeding drew the endometrium in the follicular phase , and definied by hematoxylin eosin (HE) staining . The objects were healthy child-bearing period women , their physical examination and blood chemical examination were normal , without using Hormone drug in 3 months before checken. The endometrium drawn by hysteroscope location was promptly fixed by 10% neutrality formalin, desiccation,pellucidum, dip-cera and embedded in paraffin. The samples proseed HE stain and constitution form observation to prove proliferative phase , after eliminate functional bleeding, ployp, submucous myoma etc.proceed immunohisto chemistry(IHC) stain. The experiment method of immunohistochemistry(IHC) is to detect the expression and distribution of VEGF and bFGF in each group. Concrete procedure as follow: 1.After deparaffinization the paraffin section was performed in 3% hydrogen peroxide solution (H2O2) hatching for 10 minutes to inactivate endogenous peroxidase. 2.antigen plerosis adopted boiling with electric stove .VEGF, bFGF plerosised differently for 20 minutes,15 minutes. 3. Normal goat blood blocked, hatched 10 minutes in room temperature. 4. Added uni-anti, overnight in 4℃ refrigerator. 5. added biotin making the first antibody-Ⅰ, hatching 30 minutes in 37℃. 6. added horseradish enzyme labeling chain enzyme avidin . hatching 30 minutes in 37℃. 7. VEGF show color for 3 minutes and bFGF for 2 minutes with DBA, tap water terminate sections were counter stained with hematoxylin for 5 mintutes , Hydrochloric acid alcohol differentiation 20 seconds . Alcohol desiceation , hyalined by dimethylbenzene. After sealed sheet by gum, observed it. Result: ㈠ Hysteroscope observation : Ⅰgroup : there were phanero-hyperemia , increasing raw blood vessel and bleeding like point or patch in endometrium. The massivebleeding had blood clot , blood fila suspending in uterine cavity , annular impression induced by mental "O"ring bleeding concentrated in the annular shape area , "T"ring across arm sticked on the metrawall tightly . There were angiectasia an...