| Objective:Pregnancy is such a procedure, the maternalimmunal system recognize the embryo with paternal alloantigenand produce protective immunological response until the fetal isdelivered. Fetomaternal immunal balance being destroyed,immunologic rejection result in the failure of embryo'sallotransplantation. So the abortion occurres. Recurrentspontaneous abortion is a common compilation of pregnancy,occurring in 1% of all population births, occurring in 15% of allhabitual abortions. The etiological factor of RSA involveshereditary,anatomy,embryo deformity and infection,etal.Besides the above, there are 40%-60% patients whoseetiological factor can't be explained. The study of recent yearsindicate that defective production of LIF,M-CSF and Th2-typecytokines at fetomaternal interface is associated with recurrentspontaneous abortion. There are not reports about therelationships between serum levels of LIF, M-CSF and RSA.Theaim of our study is to investigate the changes of serum IL-10,M-CSF,LIF levels after active immunotherapy in women withunexplained recurrent spontaneous abortion (RSA).We try tofind out the role IL-10,M-CSF,LIF play in RSA and active 4immunotherapy. Methods: The patients were selected from the outpatientsof Obstetrical and Gynecology, second Affiliated Hospital,Hebei Medical University. Abortion group:20 RSA patients, thenumber of abortions≥3,eliminating the factors of chromatosome,anatomy, endocrine, infection and embryo deformity .All thepatients receive active immunotherapy, venous blood wasdrawed pretherapy and three weeks after a course wascompleted. Normal non-pregnancy group:20 women whorequest to place intrauterine device, They all have a history ofdelivery, do not have a history of spontaneous abortion, norhave a history of fetal death,stillbirth. Normal pregnancygroup:20 women who request artificial abortion because ofpregnancy for 40-45 days didn't have a history of informalpregnancy nor have a history of the above diseases. They didn'thave birth sign before artificial abortion. Elbow venous bloodwas collected into a sterile tube without anticoagulant. All bloodsamples were stored at -70℃ refrigerator. Concentrations ofIL-10 , M-CSF , LIF were measured by enzyme-linkedimmunosorbant assay (ELISA) method in sera from twentycases of unexplained RSA women before and after activeimmunotherapy. Twenty normal non-pregnancy (NNP)womenand twenty normal pregnancy(NP)women were taken as control.Concrete procedure as follow:(1)Secure the desired number ofcoated wells in the holder. (2)Dispense 100μl of standard intoappropriate wells. (3) Dispense 50μl of Specimen Diluent into 5appropriate wells. (4) Dispense 50μl of Specimens intoappropriates wells. (5) Dispense 50μl of Enzyme ConjugateReagent to each well. (6)Thoroughly mix for 30 seconds. It isvery important to have a complete mixing in this setup. (7)Incubate for 60 minutes at 37℃. (8) Rinse and empty themicrotiter wells 6 times with Washing Buffer. (9) Strike thewells sharply onto absorbent paper or paper towels to remove allresidual water droplets. (10)Dispense 50μl Color Reagent A and50μl Color Reagent B into each well. (11) Incubate for 15minutes at 37℃. (12) Stop the reaction by adding 50μl of StopSolution to each well. (13) Gently mix for 30 seconds.(14)It isimportant to make sure that all the blue color changes to yellowcolor completely. (15) Read the optical density at 450nm with amicrotiter plate reader within 30 minutes. Results:(1) Serum concentrations of IL-10 and M-CSFwere lower significantly (P<0.01) in RSA women[(0.19±0.048)pg/ml and(100.99±8.32)pg/ml,respectively]than those in NNP women [ ( 0.24±0.026 ) pg/ml and(110.97±11.82)pg/ml, respectively] .However, there were nosignificant differences in serum concentrations of LIF betweenRSA women and NNP women.
|
PDF Full Text Request