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Revaluation of cytogenetic diagnosis of partial hydatidiform moles by short tandem repeats polymorphism analysis
A16
Abstracts / Placenta 34 (2013) A1–A17
generates oxidative metabolites called biopyrrins (BPns) as a result of the reaction with reactive oxygen species. Then in this study, to elucidate whether the HO-1 and BPns in human placenta were morphologically investigated. Methods: Placental tissues from ten patients complicated with PIH and seven patients with preterm birth delivery indicated by maternal disease as normal controlled were examined. All deliveries were caesarian sectioned, and no significant differences in delivery weeks were confirmed between two groups. The immunohistochemical stainings were used with anti-bilirubin (24G7) and anti-HO-1 (EP1391Y) monoclonal antibodies. Results: In the PIH group, immune-localizations of EP1391Y were demonstrated around decidual spiral arteries, especially with atherosis, in the area of infarction and villous stroma accompanied with peri-villous fibrin deposits, and in the syncytial knot. 24G7-localizations were identified around the area of infarction and decidual vasculopathy. These proteins were stained with more diffuse and high intense degree in those of PIH than those of controlled group. Conclusions: This is the first report to demonstrate the localization of BPns (24G7) immunohistochemically in human placenta not only obtained from normal controlled but PIH cases. However, few studies have reported placental HO-1 expression levels are lower in women with PIH in contrast to our result. We have to study more the BPn mechanisms in PIH cases.
http://dx.doi.org/10.1016/j.placenta.2013.07.057
No.45 REVALUATION OF CYTOGENETIC DIAGNOSIS OF PARTIAL HYDATIDIFORM MOLES BY SHORT TANDEM REPEATS POLYMORPHISM ANALYSIS Jia Qu, Hirokazu Usui, Makio Shozu Department of Reproductive Medicine, Chiba University Graduate School of Medicine, Chiba, Japan
Abstract Purpose: Partial hydatidiform moles (PM) have a triploid, diandric and monogynic genome arising from the fertilization of a normal egg by two spermatozoa. Three allelic loci of villous tissue by short tandem repeat analysis indicate triploidy or trisomy. In two allelic loci, triploidy or trisomy would be deduced from uneven peak heights, theoretically the ratio is 1:2, whereas diploid cells show the even peak heights. In this study, we tested if the measured peak-height ratio of two allelic loci could distinguish triploid from diploid. Methods: Genomic DNA was extracted from patients’ peripheral blood cells and chorionic villi. We analyzed STR polymorphism with PowerPlex16HS system compared patient and her villous tissue genotyping profiles. Fifteen cases of PM that have two or more loci with three alleles were included. We measured all peak-height ratios (peak height of shorter peak/ length of longer peak) of two allelic loci of PM and patients. We compared the distributions of peak-height ratio between PM (triploidy) and patients (diploidy). Results: Two allelic loci of 141 in PM and 177 in patients were analyzed. The mean peak-height ratios were 0.550.15 in PM and 0.860.11 in patients, respectively. Assuming that the discrimination ratio is 0.7, the two allelic loci of 119 (84%) in 141 partial moles were categorized as triploid, and the loci of 166 (94%) in 177 patients were categorized as diploid. Conclusion: This multiplex PCR STR assay could discriminate the ploidy with the peak-height ratio of two allelic loci.
http://dx.doi.org/10.1016/j.placenta.2013.07.058
No.46 PATHWAY ANALYSIS PREECLAMPSIA
OF
MICROARRAY
DATA
INVOLVED
IN
Kaoru Kawasaki, Eiji Kondoh, Yoshitsugu Chigusa, Ryusuke Murakami, Mari ujita, Hikaru Kiyokawa, Fumitomo Nishimura, Kohei Fujita, Haruta Mogami, Ikuo Konishi Department of Gynecology and Obstetrics, Kyoto University Graduate School of Medicine, Kyoto, Japan
Abstract Objective: The pathophysiology of preeclampsia has not been fully clarified. The aim of the present study was to identify the pathways related to the pathophysiology of preeclampsia by analysis of gene expression microarray profiling. Methods: Gene expression date of the preeclamptic and normal control placentas from GEO (Gene Expression omnibus) were obtained, and ssGSEA (single sample Gene Set Enrichment Analysis) was performed to assess gene set activation scores. Candidate pathways, which were upregulated or down-regulated in preeclampsia, were identified from three platforms (t-test; GSE25906: p<0.01, GSE10588: p<0.001, GSE14722: p<0.005). These selected pathways were validated using hierarchical clustering (gene cluster 3.0). RNA was purified from tissue biopsies from 10 placentas of preeclampsia and 10 normal controls. The expressions of selected genes involved in the pathways were examined by RT-PCR. Results: 24 pathways were significantly up-regulated or down-regulated in preeclampsia. The pathways were associated with cell proliferation, apoptosis, amino acid metabolism and anti-oxidative stress. Of 24, glutathione pathway, which was closely linked to anti-oxidative stress, was suppressed in preeclampsia. Indeed, the mRNA expressions of GCLC (glutathione cysteine ligase catalytic subunit) and GCLM (glutathione cysteine ligase modulatory subunit) were decreased in preeclampsia (Mann-Whitney test, GCLC; p¼0.0029, GCLM; p¼0.0005). Conclusions: The 24 candidate pathways can be closely related with the pathophysiology of preeclampsia. http://dx.doi.org/10.1016/j.placenta.2013.07.059
No.47 A CASE OF PLACENTA INCRETA IN A PATIENT WITH A HISTORY OF UAE FOR POSTPARTUM HEMORRHAGE Fuki Hirowatari
Abstract Uterine Artery Embolization (UAE) has been used to treat postpartum hemorrhage to avoid hysterectomy. A few studies have reported the pregnancy outcome after this procedure. [Case] A 29-year-old primigravida had transferred to our hospital because of a risk of preterm delivery at 34 weeks. She had normal vaginal delivery soon after the arrival followed by increasing postpartum hemorrhage. She had undergone bilateral UAE to control the bleeding. 4 years and 5 months later, she became pregnant naturally, and the course of pregnancy was uneventful except breech presentation. During scheduled cesarean section, we found several abnormal extended vessels and placenta around 3x4cm square were invading the myometrium reaching on the uterine surface serosa. We inverted the uterus and removed the placenta manually resulting in missing some part of myometrium because it was firmly adherent. We left uterus in situ, not having the authorization for an hysterectomy. 7 days after the operation，she had fever and abscess resisting antibiotic therapy. Supra-cervical hysterectomy was performed 28 days after the first operation and she was discharged home on day 41.UAE is widely used to treat various conditions for the management of postpartum hemorrhage. But
Abstracts / Placenta 34 (2013) A1–A17
generates oxidative metabolites called biopyrrins (BPns) as a result of the reaction with reactive oxygen species. Then in this study, to elucidate whether the HO-1 and BPns in human placenta were morphologically investigated. Methods: Placental tissues from ten patients complicated with PIH and seven patients with preterm birth delivery indicated by maternal disease as normal controlled were examined. All deliveries were caesarian sectioned, and no significant differences in delivery weeks were confirmed between two groups. The immunohistochemical stainings were used with anti-bilirubin (24G7) and anti-HO-1 (EP1391Y) monoclonal antibodies. Results: In the PIH group, immune-localizations of EP1391Y were demonstrated around decidual spiral arteries, especially with atherosis, in the area of infarction and villous stroma accompanied with peri-villous fibrin deposits, and in the syncytial knot. 24G7-localizations were identified around the area of infarction and decidual vasculopathy. These proteins were stained with more diffuse and high intense degree in those of PIH than those of controlled group. Conclusions: This is the first report to demonstrate the localization of BPns (24G7) immunohistochemically in human placenta not only obtained from normal controlled but PIH cases. However, few studies have reported placental HO-1 expression levels are lower in women with PIH in contrast to our result. We have to study more the BPn mechanisms in PIH cases.
http://dx.doi.org/10.1016/j.placenta.2013.07.057
No.45 REVALUATION OF CYTOGENETIC DIAGNOSIS OF PARTIAL HYDATIDIFORM MOLES BY SHORT TANDEM REPEATS POLYMORPHISM ANALYSIS Jia Qu, Hirokazu Usui, Makio Shozu Department of Reproductive Medicine, Chiba University Graduate School of Medicine, Chiba, Japan
Abstract Purpose: Partial hydatidiform moles (PM) have a triploid, diandric and monogynic genome arising from the fertilization of a normal egg by two spermatozoa. Three allelic loci of villous tissue by short tandem repeat analysis indicate triploidy or trisomy. In two allelic loci, triploidy or trisomy would be deduced from uneven peak heights, theoretically the ratio is 1:2, whereas diploid cells show the even peak heights. In this study, we tested if the measured peak-height ratio of two allelic loci could distinguish triploid from diploid. Methods: Genomic DNA was extracted from patients’ peripheral blood cells and chorionic villi. We analyzed STR polymorphism with PowerPlex16HS system compared patient and her villous tissue genotyping profiles. Fifteen cases of PM that have two or more loci with three alleles were included. We measured all peak-height ratios (peak height of shorter peak/ length of longer peak) of two allelic loci of PM and patients. We compared the distributions of peak-height ratio between PM (triploidy) and patients (diploidy). Results: Two allelic loci of 141 in PM and 177 in patients were analyzed. The mean peak-height ratios were 0.550.15 in PM and 0.860.11 in patients, respectively. Assuming that the discrimination ratio is 0.7, the two allelic loci of 119 (84%) in 141 partial moles were categorized as triploid, and the loci of 166 (94%) in 177 patients were categorized as diploid. Conclusion: This multiplex PCR STR assay could discriminate the ploidy with the peak-height ratio of two allelic loci.
http://dx.doi.org/10.1016/j.placenta.2013.07.058
No.46 PATHWAY ANALYSIS PREECLAMPSIA
OF
MICROARRAY
DATA
INVOLVED
IN
Kaoru Kawasaki, Eiji Kondoh, Yoshitsugu Chigusa, Ryusuke Murakami, Mari ujita, Hikaru Kiyokawa, Fumitomo Nishimura, Kohei Fujita, Haruta Mogami, Ikuo Konishi Department of Gynecology and Obstetrics, Kyoto University Graduate School of Medicine, Kyoto, Japan
Abstract Objective: The pathophysiology of preeclampsia has not been fully clarified. The aim of the present study was to identify the pathways related to the pathophysiology of preeclampsia by analysis of gene expression microarray profiling. Methods: Gene expression date of the preeclamptic and normal control placentas from GEO (Gene Expression omnibus) were obtained, and ssGSEA (single sample Gene Set Enrichment Analysis) was performed to assess gene set activation scores. Candidate pathways, which were upregulated or down-regulated in preeclampsia, were identified from three platforms (t-test; GSE25906: p<0.01, GSE10588: p<0.001, GSE14722: p<0.005). These selected pathways were validated using hierarchical clustering (gene cluster 3.0). RNA was purified from tissue biopsies from 10 placentas of preeclampsia and 10 normal controls. The expressions of selected genes involved in the pathways were examined by RT-PCR. Results: 24 pathways were significantly up-regulated or down-regulated in preeclampsia. The pathways were associated with cell proliferation, apoptosis, amino acid metabolism and anti-oxidative stress. Of 24, glutathione pathway, which was closely linked to anti-oxidative stress, was suppressed in preeclampsia. Indeed, the mRNA expressions of GCLC (glutathione cysteine ligase catalytic subunit) and GCLM (glutathione cysteine ligase modulatory subunit) were decreased in preeclampsia (Mann-Whitney test, GCLC; p¼0.0029, GCLM; p¼0.0005). Conclusions: The 24 candidate pathways can be closely related with the pathophysiology of preeclampsia. http://dx.doi.org/10.1016/j.placenta.2013.07.059
No.47 A CASE OF PLACENTA INCRETA IN A PATIENT WITH A HISTORY OF UAE FOR POSTPARTUM HEMORRHAGE Fuki Hirowatari
Abstract Uterine Artery Embolization (UAE) has been used to treat postpartum hemorrhage to avoid hysterectomy. A few studies have reported the pregnancy outcome after this procedure. [Case] A 29-year-old primigravida had transferred to our hospital because of a risk of preterm delivery at 34 weeks. She had normal vaginal delivery soon after the arrival followed by increasing postpartum hemorrhage. She had undergone bilateral UAE to control the bleeding. 4 years and 5 months later, she became pregnant naturally, and the course of pregnancy was uneventful except breech presentation. During scheduled cesarean section, we found several abnormal extended vessels and placenta around 3x4cm square were invading the myometrium reaching on the uterine surface serosa. We inverted the uterus and removed the placenta manually resulting in missing some part of myometrium because it was firmly adherent. We left uterus in situ, not having the authorization for an hysterectomy. 7 days after the operation，she had fever and abscess resisting antibiotic therapy. Supra-cervical hysterectomy was performed 28 days after the first operation and she was discharged home on day 41.UAE is widely used to treat various conditions for the management of postpartum hemorrhage. But