| Backgroundand ObjectiveIn recent years,recurrent spontaneous abortion(RSA)has gradually become one of the common clinical diseases in gynecology.The World Health Organization(WHO)defines RSA as a spontaneous abortion occurring three or more times before the 20th week of pregnancy.As the incidence of RSA rises year by year,some experts suggest that spontaneous abortion of twice or more should be highly assessed and intervened to avoid RSA.The causes of RSA are complicated,mainly include genetic factors,immune dysfunction,endocrine factors,anatomical factors,infectious factors,prethrombotic state,etc.Therefore,it is absolutely necessary to clarify the cause to assist in diagnosis and treatment.With the continuous exploration of the causes of RSA,embryo chromosomal abnormalities is one of the important causes,people with autoimmune disorders have a high risk of abortion,and old age is also one of its high risk factors.This study examined the chromosomes of abortive tissue embryos,compared and analysed the value of the next generation sequencing technology in the genetics of recurrent spontaneous abortion,studied the relevant factors of recurrent spontaneous abortion,hope to provide basis for the prevention and treatment of RSA.Materials and Methods1 Materials248 cases of spontaneous abortion from September 2013 to December 2017 in the Second Affiliated Hospital of Zhengzhou University were collected.Chorionic celles of their abortion tissues were cultured and then examined by G-banding karyotype analysis.73 cases of spontaneous abortion from January 2015 to November 2017 in this hospital were collected.Chorionic celles of their abortion tissues were examined by next generation sequencing(NGS)technology.According to the age of pregnant women,the overall study was divided into groups of?35 and<35.According to the number of spontaneous abortions of pregnant women,the subjects were divided into spontaneous abortion group 3 times and spontaneous abortion group?4 times.There were no significant difference in the number of births,parity and body mass index between the both groups(P>0.05).Reviewed the medical records and followed up the chromosomes and autoimmune results of all pregnant couples at the same time.Enrollment criteria:(1)Spontaneous abortion patients who self-discharged pregnancy tissue by vaginal or clinically diagnosed as missed abortion.(2)Number of spontaneous abortions(including this pregnancy)?3times.(3)Exclusion of genital tract organic disease by ultrasound.(3)Exclusion of genital tract infections by testing cervical secretion.2 Statistic AnalysisUsing SPSS17.0 software to analyze the data.Measurement datas were described with the mean±standard deviation(?x±s).To use paired t test analysed the treatment outcome in each group.Chi-square test is for counting data which expressed as a percentage.There were significant differences as P<0.05.Results1 Villus chromosome results of abortion embryonic tissuesTwo of the 248 cases of chorionic villus cell culture and chromosome G banding karyotype analysis techniques failed to culture,and the rest were successfully detected with a success rate of 99.2%.134 cases were normal in chromosome of G-banding karyotype analysis in 246 cases,accounting for 54.5%.103 cases were abnormal in chromosome number(including 28 cases of chimeras),accounting for41.9%.9 cases were abnormal in chromosome structure,accounting for 3.7%.All of73 specimens tested by NGS were successful,the rate of success was 100%.28 cases were abnormal in chromosome,accounting for 38.4%.19 cases were abnormal in chromosome number,accounting for 26.0%.26 cases were abnormal in chromosome structure,accounting for 35.6%.2 Peripheral blood chromosome test results of both parents of aborted embryos163 peripheral blood chromosome results(the both parents)were collected according to view medical records and follow-up the patients.There were 4 cases of maternal chromosome abnormalities,including 3 cases of chromosome balance translocation and 1 case of inversion karyotype in arm 10.The fathers'peripheral blood were normal.3 Age factor319 subjects were divided into?35-year-old group and<35-year-old group according to the age of the pregnant women,comparing the percentage of villous chromosomal abnormalities in two groups.The rate of villous chromosomal abnormalities(57.6%)was higher in those aged?35 years than in those who were<35 years old(40.3%),?~2=9.556,P=0.002.In the two groups,the number of abnormal chromosomes were dominated by three bodies,accounting for 48.1%(38/79)and39.5%(17/43)respectively.Comparison of the positive rates of autoimmune antibodies between the two groups was not statistically significant,P=0.891.4 The relationship between the number of spontaneous abortion with chromosomal abnormalities and autoimmune antibody positive319 subjects were divided into spontaneous abortion of 3 times and?4 times according to the number of spontaneous abortions,comparing the rate of chorionic chromosomal abnormalities in two groups.The rate of villous chromosomal abnormalities in patients who had spontaneous abortion of 3 times(63.4%)was higher than that of?4 times(31.9%),?2=31.330,P=0.000.In the two groups,the number of abnormal chromosomes was mainly in three-body,accounting for 46.0%(40/87)and 42.9%(15/35)respectly.Compared the positive rates of autoimmune antibodies in the two groups,the rate of autoimmune antibody positive was higher in spontaneous abortion patients of 3 times(49.3%)than that of?4 times(33.7%),?2=3.902,P=0.048.Conclusions1 NGS has higher chromosome detection success rate and chromosome microdeletions/microreplications structural abnormalities detection rate than villus cell culture and chromosome G banding karyotype analysis techniques.2 Patients with RSA should pay attention to the chromosome examination of both the embryo and the parents in order to clarify the cause of abortion and provide genetic counseling and recommendations for repregnancy.3 Age?35 years is a highly risk factor for abnormal chromosomes in embryos that cause RSA.4 The abnormal chromosome rate of embryos decrease and the positive rate of autoimmune antibodies increase in RSA patients with the increasing of the number of abortion. |
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