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Objective To investigate the features and characteristics of GLIS3 gene mutation in patients with congenital hypothyroidism(CH),and to establish the theoretical basis for gene diagnosis and prenatal diagnosis of CH.Methods Genomic DNA was extracted from peripheral blood leukocytes of 50 patients with CH who were collected from February 2007 to November 2016 in Shandong Province.The exon 2 to 11 of GLIS3 were amplified with 11 pairs of sequence specific primers designed by Primer 5.0.Polymerase chain reaction and the first generation of sequencing method(Sanger sequencing) were used to detect the mutation.Comparison of the sequencing results with the GLIS3 reference sequence (National Center for Biotechnology Information Reference Sequence:NC_000009.12) helped to screen gene mutations.Results The 50 CH patients included 22 boys and 28 girls,and the sex ratio was 1.0 ∶ 1.3.The mean age was (2.5 ± 0.5) years.Six cases (12％) had thyroid gland hypoplasia,23 cases (46％) had thyroid gland agenesis and 21 cases(42％) with ectopic thyroid gland.C2507A missense mutation was found in exon 10 of GLIS3 in a thyroid gland agenesis case,which might result in proline to glutamine substitution at codon 836.One mutant (rs780019691,c.C289T) was detected which was nonsense mutation (Arg→Stop) in another thyroid gland agenesis child.Conclusions The mutation rate of GLIS3 gene is very low in CH children of Shandong province.Further studies are needed to investigate the relationship between GLIS3 genotypes and clinical phenotypes.
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Objective To investigate the relationship of S100B protein expression and the pathogenesis of early-onset and late-onset preeclampsia.Methods Sixty patients with preeclampsia who received caesarean section at Qingdao Municipal Hospital from October 2010 to September 2011 were enrolled in this study.Thirty cases were early-onset preeclampsia( referred as early-onset preeclampsia group,＜ 34 weeks),and the other 30 cases were late-onset preeclampsia (referred as late-onset preeclampsia group,≥34 weeks).Thirty women who received caesarean section because of pelvic structural deformities,breech presentation,macrosomia and social factors were included as the control group.The expression of S100B mRNA in the placenta was detected by reverse transcription ( RT)-PCR.The expression of S100B protein in the placenta was detected by immunohistochemistry.Results ( 1 ) S100B mRNA was expressed in the trophoblasts of preeclampsia and control groups.The expression of S100B mRNA in early-onset preeclampsia group (0.73 ±0.11 ) was significantly higher than the control group (0.58 ±0.08) and lateonset preeclampsia group (0.64 ±0.10,P ＜0.05 ).There was no significant difference between late-onset preeclampsia group and the control group ( P ＞ 0.05 ).(2) S100B protein was expressed in the plasma membrane and cytoplasm of the trophoblasts,correlated positively with the brownish yellow and brown particles inside the cells.It was expressed in all the three groups.Immunohistochemistry revealed that the expression of S100B protein in the placenta of early-onset preeclampsia group was 100％ (30/30),significantly higher than those of late-onset preeclampsia group and the control group,in which the positive rate were 70％ (21/30) and 63％ (19/30) respectively (P ＜0.05).There was no difference between late onset preeclampsia group and the control group (P ＞0.05).Conclusion Early-onset and late-onset preeclampsia may have different etiology and pathogenesis.S100B may be a factor in the pathogenesis of early-onset preeclampsia.