| Part I Study on the expression of BDNF gene in placenta of preeclampsiaObjective: By analyzing the gene expression of BDNF in placenta tissue collected from women with preeclampsia,explore the possible relationship between BDNF and preeclampsia.Methods: 21 cases of women with preeclampsia were collected from the Affilited Hospital of Qingdao University during June 2017 to June 2018 as case group(PE group),and 21 cases in normal pregnant as control group(control group).RNA and protein were extracted from placentas of 42 targets to detect expression levels of BDNF by RT-PCR and Western-blot.Results: The expression level of BDNF in PE group was lower than that in normal control group(T=2.337,P=0.0245)at mRNA level.At the protein level,which was detected by WB and image analysis,expression of BDNF in PE group was significantly lower than that in control group,and the difference was statistically significant(T=3.531,P=0.0011).Conclusion: The BDNF gene has a low expression trend in the placenta of patients with preeclampsia,which may result in preeclampsia,and BDNF may play a protective role in this disease.Part II Polymorphism analysis of Val66Met in BDNF gene of preeclampsia patientsObjective: Analyze the relationship between the single nucleotide polymorphism of BDNF Val66Met and the genetic susceptibility of preeclampsia.Methods: Targets of study were from the preeclampsia sample pool established by the Prenatal Diagnostic Laboratory of the Affiliated Hospital of Qingdao University during January 2014 to December 2017.The R i386 3.4.3 software was used to eliminate and match the relevant factors and obtained 1138 cases as case group from 1800 cases and 1342 cases as control group from 5000 cases.DNA was extracted from peripheral blood of all subjects and single nucleotide polymorphism was analyzed by Taq Man probe fluorescent PCR method,and 30 samples were randomly selected from all samples to be analyzed by first generation sequencing to verify the accuracy of the results obtained with the probe besides.Results: 1.The clinical data showed that there was no different between case group and control group in maternal age,BMI,pregnancy times and number of births(P>0.05).Maternal blood pressure of case group,including systolic blood pressure and diastolic blood pressure,was significantly higher than that in control group(P<0.05).Gestational weeks of admission and delivery and birth weight of newborns in case group were lower than those in control group(P <0.05).2.Compared with control group,the frequency distribution of Val66Met alleles of BDNF in case group was statistically different(X2=4.657,P=0.031,OR=0.884,95%CI=0.791-0.989).There was no statistical difference in genotype distribution analysis(X2=4.555,P=0.103),and there was no statistical difference in additive gene too(X2=2.682,P=0.105,OR=0.856,95%CI=0.710-1.031).Conclusions: 1.Preeclampsia leads to earlier time of maternal need for medical intervention and even delivery.2.The single nucleotide polymorphism of BDNF gene of the maternity,Val66Met,may be associated with preeclampsia genetic susceptibility.Part III Analysis of single nucleotide polymorphism of BDNF locus Val66Met in mother fetal pairing and association between preeclampsia and neonatal outcomesObjective: By analyzing the clinical data of neonates delivered by mothers with preeclampsia,the relationship between preeclampsia and neonatal adverse outcomes was explored.In addition,the inheritance and transmission of Val66Met of BDNF between maternal and fetal were explored by analyzing peripheral DNA.Methods: From January 2018 to June 2018,targets were amassed from the neonatology department of the Affiliated Hospital of Qingdao University,consisting of 200 infants delivered by mother with preeclampsia as case group and 208 infants delivered by healthy parturient as control group.The clinical data of the infants and mothers were collected and were retrospectively analyzed in gestational age matching groups(<30 weeks,30~33+6weeks and 34~36+6 weeks)to explore the relationship between preeclampsia and neonatal adverse outcomes.At the same time,Taq Man probe fluorescent PCR was used to analyze the allele frequency and genotype distribution of Val66Met in maternal fetal pairing.20 cases were randomly selected for sequencing to verify the accuracy of the test results.Results: 1.There was no statistical difference in maternal pregnancy and parity times between two groups(P>0.05).The birth weight,birth gestational age and 1-minute Apgar score of case group were statistically lower than those of control group(P<0.05).2.Compared with the control group,incidence rate of Small for Gestational Age and Bronchopulmonary Dysplasia were statistically higher in the <30 weeks gestational age group and 30~33+6 gestational age group(P<0.05).In the 34~36+6 gestational age group,the incidence of Small for Gestational Age was still higher than that of control group(P<0.05).There was no statistical difference in other diseases such as Respiratory Distress Syndrome,Patent Ductus Arterious,White Matter Damage,intracranial hemorrhage and Necrotizing Enterocolitis between two groups after gestational age matched(P>0.05).3.The alleles frequency distribution of Val66Met in BDNF was statistically different in mothers(X2=9.054,P=0.003,OR=0.630,95%CI=0.466-1.852),consistent with the results of large sample data,and the genotype and additive genes were not statistically different(P>0.05).However,there was no statistical difference between two groups in genotype distribution,allele and additive gene analysis of fetal Val66Met(P>0.05).In addition,the analysis of Val66Met transfer in maternal-fetal pairing showed that there was no statistical difference between case group and control group(X2 = 4.896,P = 0.086).Conclusions: 1.Preeclampsia leads to intrauterine growth restriction in infants,and may affect fetal lung development,resulting in an increase in the incidence of Small for Gestational Age and Bronchopulmonary Dysplasia.2.Single nucleotide polymorphisms of Val66Met in BDNF gene may be not associated with preeclampsia-related neonatal prognosis. |
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