Detection of Homozygous Deletions and Mutations in the CDKN2A Gene in Hydatidiform Moles

OBJECTIVE To investigate homozygous deletions and mutations in the CDKN2A gene(p16 INK4a and p14 ARF gene)in hydatidiform moles. METHODS A total of 38 hydatidiform mole samples and 30 villi samples were examined for homozygous deletions in the CDKN2A gene by PCR and for mutations by DHPLC. RESULTS i)Among 38 hydatidiform mole samples, homozygous deletions in the p16 INK4a exon 1 were identified in 5 cases(13.2%),while no homozygous deletions were found in the p16I NK4aexon 1 of 30 early-pregnancy samples.The rates of those deletions in hydatidiform compared to early-pregnancy villi samples was statistically significant(P=0.036).ii)No homozygous deletions in the p14 ARF exon 1 or p16 INK4a exon 2 were found in any of the hydatidiform moles or early-preganancy samples.iii) In all hydatidiform moles and early-pregnancy villi samples,no mutations were detected by DHPLC. CONCLUSION We suggest there may be a close correlation between homozygous deletions in the CDKN2A gene and occurrence of hydatidiform moles variation in the CDKN2A gene is mainly caused by homozygous deletions,while mutations may be not a major cause.

Single Nucleotide Polymorphism-Based Chromosomal Microarray Evaluation of Hydatidiform Moles: A US National Reference Laboratory Experience

Objectives: This retrospective study evaluated 1) benefits of single nucleotide polymorphism (SNP)-based chromosomal microarrays (CMAs) in the diagnosis of complete hydatidiform mole (CHM) and partial HM (PHM) in products of conception (POC) and amniotic fluid (AF) specimens, and 2) frequency of whole-genome uniparental disomy (wgUPD) and triploidy in POC and AF specimens received at a US national reference laboratory. Methods: We reviewed consecutive 2138 POC and 3230 AF specimens and identified the cases with wgUPD and triploidy which are associated with molar pregnancy. Results: Of 2138 consecutive POC specimens tested, SNP-based CMA detected wgUPD in 10 (0.47%) and triploidy in 84 (3.93%). Of the 10 wgUPD cases, 9 (90%) were confirmed as CHM. Of 3230 consecutive AF specimens, the array detected wgUPD in 1 case (0.03%) and triploidy in 11 (0.34%). Conclusions: SNP-based microarray allows detection of wgUPD in POC and AF specimens at a US national reference laboratory. Correctly diagnosing HM and differentiating CHM from PHM are important for clinical management. The effective SNP-based CMA detection of wgUPD in CHM may enable physicians to monitor patients at risk for gestational trophoblastic disease and neoplasm. Conventional chromosome analysis of POC has a high failure rate, cannot be performed on formalin-fixed paraffin embedded samples, and cannot detect wgUPD. Further multi-institutional collaborative assessmen on accuracy, cost-effectiveness, and adequate access to SNP-based CMA, may lead this testing platform to be considered as the first-tier analysis tool for POC specimens, including those showing PHM or CHM.

Complete Hydatidiform Mole at Menopause: A Case Report from Panzi Hospital, Bukavu, Democratic Republic of Congo

Gestational trophoblastic disease is an abnormal proliferation of trophoblastic tissue during pregnancy. It occurs in women of childbearing age, although a few cases have also been observed in post-menopausal women, although it is extremely rare in the latter. Here we describe a rare case of complete hydatidiform mole in a 56-year-old female patient who presented with genital bleeding combined with nausea and vomiting and a gravid uterus 16 cm in height. The ultrasound findings and the increase in serum β-HCG to 182566.00 mIU/ml suggested a diagnosis of complete hydatidiform mole. Given the post-menopausal state and the future risk of post-molar gestational trophoblastic neoplasia, we opted for total hysterectomy without preservation of the adnexa via a transabdominal approach, followed by antimitotic treatment with methotrexate. The uterus measured 18.45 cm × 11.18 cm with intra cavitary vesicles. Microscopic examination showed chorionic villi of variable size and shape, most of which were dilated and oedematous, associated with trophoblastic cell proliferation and haemorrhage suggestive of complete benign hydatidiform mole. Follow-up showed a consistent decrease in serum β-HCG levels and no evidence of residual disease. A suspicion of gestational trophoblastic disease should be borne in mind when evaluating a patient with peri- or post-menopausal bleeding to avoid delay in diagnosis and treatment.

Genetically identified complete hydatidiform mole coexisting with a normal fetus following IVF-ET

A complete hydatidiform mole coexisting with a fetus following in vitro fertilization and embryo transfer (IVF-ET) is a rare event. The diagnosis is often not easy because of the morphological similarity to a partial mole, but important to the treatment. We present a recent case in which STR polymorphism analysis clearly revealed a different genetic origin for the fetal and molar parts. STR polymorphisms on 15 variable number tandem repeat loci and a gender-determination locus, which were detected by polymerase chain reaction, indicating that the cord/placenta and molar tissue were parental and androgenous, respectively. During follow-up, the patient developed persistent gestational trophoblastic tumor (GTT) which was successfully treated with chemotherapy. In this case, STR polymorphism analysis exactly diagnosed a twin pregnancy consisting of a complete hydatidiform mole and a fetus.

Study on neoplasia of hydatidiform mole by detecting telomerase reverse transcriptase mRNA expression in peripheral blood mononuclear cells

By setting up a real-time fluorescent quantitative RT-PCR assay to detect human telomerase reverse transcriptase （hTERT） mRNA in hydatidiform mole in peripheral blood mononuclear cells, to analyze the correlation between the expression level of hTERT mRNA and the prognosis of hydatidiform mole, and to evaluate the clinic value of quantitative determination of hTERT mRNA in the diagnosis of hydatidiform mole. Methods： A real-time fluorescent quantitative RT-PCR （FQ RT-PCR） assay based on TaqMan fluorescence methodology and the Light-Cycler system was used to quantify the full range of hTERT mRNA copy numbers in 30 samples of hydatidiform mole and the neoplasia of hydatidiform mole. The normalized hTERT （NhTERT） was standardized by quantifying the number of GAPDH transcripts as internal control and expressed as 100x （hTERT/GAPDH） ratio. Based on the prognosis of the hydatidiform mole, the patients were divided into two groups： the experimental group and the control group, to compare the telomerase reverse transcriptase mRNA expression in peripheral blood mononuclear cells. Results： hTERT mRNA was both expressed in the peripheral blood mononuclear cells and pathological tissues in the mole experimental group and the control group. In the mole experimental group, the values were 6.31±0.32 and 6.24±0.44, respectively, and there was no significant difference between them （P〉0.05）. But in the control group the values were 1.21±0.65 and 1.40±0.61, respectively, and there was no significant difference between them （P〉0.05） The values in experimental group was significantly higher than those in the control group （P〈0.01）. Conclusion： Quantitative determination of hTERT mRNA by FQ RT-PCR is a rapid and sensitive method, hTERT in peripheral blood mononuclear cells may have potential use as a biomarker for the early detection of the prognosis of the hydatidiform mole.

Preliminary Study on c-Ha-ras Oncogene Mutations in Hydatidiform Mole Tissues

ve To study the presence of c-Ha-ras oncogene mutations in hydatidiform mole (HM) tissues and to further explore its relationship with mole's malignancy

Low and Maternal-specific Expression of p57^(KIP2) in Hydatidiform Mole and Its Clinical Implication

Summary: In situ hybridization was applied to locate and detect the expression of p57 KIP2 in hydatidiform mole (5 cases of partial hydatidiform mole and 18 cases of complete hydatidiform mole) and normal villi (23 cases). The positive signals of p57 KIP2 expression were analyzed by HPIAS-1000 Image-Analysis System. p57 KIP2 was highly expressed in normal villi but showed distinct low expression in hydatidiform mole (P<0.01). Furthermore, the locus of low expression of p57 KIP2 accorded with the place where lesion of trophoblast occurred. Detection of p57 KIP2 made it possible to study the genetics of hydatidiform mole at the transcriptional level. Low expression of p57 KIP2 could be a molecular marker in hydatidiform mole and a target for therapy.

Special epithelioid trophoblastic tumor:A case report

BACKGROUND Epithelioid trophoblastic tumor(ETT)is a special type of gestational trophoblastic tumor.However,its pathogenesis has been incompletely elucidated.ETT rarely occurs in the ovaries and fallopian tubes,unlike placental site trophoblastic tumor,requiring a histopathological biopsy and immunohistochemistry for further diagnosis.CASE SUMMARY A 29-year-old woman with irregular vaginal bleeding and elevated serum chorionic gonadotropin(β-hCG)levels presented similar symptoms to ectopic pregnancy.Transvaginal ultrasound revealed abnormal echoes of the left adnexa.Postoperatively,the pathology of the left ovary and fallopian tube was reported as ETT.The patient was followed up with regular hCG measurements and ultrasounds.The blood hCG values showed an upward trend 3 mo after the operation and then chemotherapy was prescribed.The current health status is normal.CONCLUSION For women of childbearing age with elevated serumβ-hCG levels,practitioners should consider ETT and be alert to the poor prognosis of the disease.After surgery,the patient’s condition should be closely observed to prevent recurrence and metastasis.Postoperative chemotherapy is only helpful for treating the disease to a certain extent.

The value of PAPP-A in the diagnosis and prognosis of GTD

Objective： To investigate the value of PAPP-A （pregnancy assouated plasma protein-A） in the diagnosis and prognosis of gestational trophoblastic disease （GTD）. Methods： The serum 13-HCG （β-human chorionic gonadotropin） and PAPP-A levels of 25 normal pregnant women, 28 patients with complete hydatidiform mole and 38 patients with invasive mole were measured by enzyme linked immunosorbent assay （ELISA） during the periods of diagnosis, treatment and follow-up. Results： Compared with control group, patients with complete mole and invasive mole had higher levels of β-HCG （P 〈 0.01）. But there was no significant difference between the complete and invasive mole group （P 〉 0.05）. The PAPP-A level of complete mole group was significantly higher than that of control group （P 〈 0.01）. The PAPP-A level of invasive mole group was significantly higher than that of complete mole group and control group（P 〈 0.05）. In complete mole group, serum β-HCG and PAPP-A levels of the patients with malignant sequelae were significantly higher than those with benign sequelae （P 〈 0.05）. The β-HCG level had no relationship with the clinical stage of invasive mole. However, the PAPP-A level increased with clinical advancement of invasive mole. The levels of β-HCG and PAPP-A gradually decreased after evacuation in patients with complete moles, but maintained positive or even increased in patients with subsequent malignancy diagnosis of hydatidiform mole and invasive mole, but Conclusion： The PAPP-A level can give us some help not only in early also in the prognosis of malignant sequelae.

Ectopic Tubal Molar Pregnancy: A Case Report

Background: Molar pregnancy is a rare condition;its ectopic form is even rarer occurring in 1.5 per 1,000,000 pregnancies. Aim: We report a case of ectopic molar pregnancy located in the fallopian tube and draw attention to the importance of systematic histological examination in any ectopic pregnancy. Case report: A 32-year-old primigravida woman presented with metrorrhagia in a context of about 2 months of amenorrhea. An ectopic pregnancy was diagnosed by ultrasonography. A Laparotomy was performed and a ruptured left ampullary ectopic pregnancy was found, and a total left salpingectomy was performed. The pathologic examination and the histological examination show an aspect of a partial molar ectopic tubal pregnancy. Conclusion: Ectopic molar pregnancy is a rare entity, its diagnosis requires histological confirmation. Beta-HCG monitoring is the essential tool for monitoring the postoperative evolution.

Twin pregnancy and partial hydatidiform mole following in vitro fertilization and embryos transfer： a novel case of placental mosaicism

Twin pregnancy with mosaic partial hydatidiform mole （PHM） and survival of two healthy fetuses following in vitro fertilization and embryos transfer （IVF-ET） is a rare situation and is considered a challenge for management. A 32-year-old Chinese woman conceived twin pregnancy following IVF-ET. At 22 weeks＇ gestation, an additional intrauterine echogenic mass with features of PHM were shown by successive ultrasound examinations. At 35 weeks＇ gestation, two live male infants and two placentas were delivered by caesarean section （CS）. Histologic examination of the abnormal placenta confirmed mosaic PHM. Genetic study showed the abnormal placental mosaicism （expressed in molar-69XXY and normal vili-46XY）, co-existing with a hypospadia new-born （46XY） in one amniotic sac. However, the other one was normal. Serial serum β-hCG levels showed a declining trend and serum β-human chorionic gonadotropin （hCG） were undetectable at 6 months after delivery. The case demonstrated that it is possible to prolonged gestation by PHM under close surveillance during the entire pregnancy.

Pulmonary deportation of hydatidiform mole:a 12-year,single tertiary center experience in China

Background:Pulmonary deportation of hydatidiform mole is an exceedingly rare entity.The underlying mechanisms and proper management strategies remain unclear based on sporadic case reports over the past six decades.This study aimed to investigate the clinical features and rational treatment of patients with benign molar pregnancies with pulmonary deportation based on our experience.Methods:Medical records of 20 cases of hydatidiform mole with pulmonary deportation were retrospectively reviewed at Peking Union Medical College Hospital from November 2006 to May 2019.The detailed information of all patients was recorded and analyzed.Patients were divided into different groups according to their characteristics and Mann-Whitney U test was used to compare the duration to achieve a normal b-human chorionic gonadotrophin(b-hCG)level after the first evacuation among groups.Results:Initial pulmonary computed tomography scans showed suspected bilateral,left and right chest deportation of hydatidiform mole in 12,four,and four patients,respectively,with the maximum nodular diameter ranging from 0.6 to 1.2 cm.Ten patients achieved lesion resolution while the remaining ten patients achieved decreases in the size of their pulmonary lesions.The median duration to achieve a normal b-hCG level after the first evacuation was 15.5(13.0,21.9)weeks.There was no significant difference in the duration to achieve a normal b-hCG level after the first evacuation between two groups based on age(≥40 years vs.<40 years:15.8[12.2,21.5]weeks vs.15.5[12.9,23.0]weeks,Z=0.094,P=0.925),type of antecedent mole(partial mole vs.complete mole:15.2[12.5,27.4]weeks vs.15.9[12.9,21.5]weeks,Z=0.165,P=0.869),distribution of pulmonary nodules(bilateral lungs vs.unilateral lung:15.2[12.8,22.5]weeks vs.15.9[13.2,22.2]weeks,Z=0.386,P=0.700),maximum size of pulmonary nodules(>0.5 cm vs.0.5 cm:13.0[11.3,17.2]weeks vs.16.0[14.5,23.8]weeks,Z=1.815,P=0.070),and number of uterine evacuations(once vs.twice or three times:15.0[13.0,16.3]weeks vs.16.0[12.8,23.9]weeks,Z=0.832,P=0.405).The post-molar cohort was followed up for 17 to 139 months,and no gestational trophoblastic neoplasia was observed.Conclusions:No surgeries other than uterine evacuation and no chemotherapy regimens are recommended for such patients if they achieve satisfactory decreases in the level of hCG and gradual decrease or disappearance of pulmonary deportation nodules.Patients should be informed about the necessity of long-term follow-up.More collaborative international studies on this exceedingly rare condition may guide decisions regarding optimal management strategies.

Analysis of Clinicopathologic Classification Features of Hydatidiform Mole Misdiagnosed as Missed Abortion Preoperatively

Objective:To analyze clinicopathologic classification features of the cases that were diagnosed as missed abortion preoperatively but turn out to be hydatidiform mole(HM)postoperatively.Methods:A retrospective analysis was conducted on the patients who had a missed abortion in our hospital from 2017 to 2018.Clinical and pathological characteristics of different types of HMs were observed.Diagnostic value of karyotype in HM was discussed based on the karyotype analysis of villi chromosome.Results:A total of 278(11.2%)HM patients were misdiagnosed as missed abortion.Naked-eye detection rate of HM was 26.61%,and sensitivity of transvaginal ultrasound on HM was 7.91%.One hundred and forty-seven(52.88%)HM cases could not be genotyped via pathological hematoxylin and eosin(HE)staining.70 HM patients underwent P57 immunohistochemistry,which had guiding significance to the classification.In addition,the karyotype diagnosis of the tissues from 15 cases basically matched their P57 classifications.Conclusions:P57 should be listed as a routine test in hydatidiform pathological examination at the same time of HE staining,and what’s more,P57 expression is consistent with genotyping,which should be recommended for the patients with HM if observed by naked eye.

Analysis of p53 expression in partial hydatidiform mole and hydropic abortion

BACKGROUND： Gestational trophoblastic disease （GTD） is a heterogeneous group of disorders characterized by abnormal trophoblast tissue. Molar and non-molar hydropic placental changes are the most common forms of GTD. Differential diagnosis of GTD is sometimes problematic. Recently, p53 expression was identified as a good marker for distinguishing GTD types. ALMS： Comparison of p53 expression in partial hydatidiform mole （PHM） and hydropic abortion. METHODS： In this prospective cross-sectional study, molar and non-molar hydropic pregnancy specimens were collected. Immunohistochemical staining, based on the Labeled Streptavidin Biotin （LSAB） technique, was carried out on multiple 4 Ixm paraffin block sections prepared from formalin-fixed trophoblastic tissues. Polymer-based Envision was used to assess p53 tumor suppressor protein immunoreactivity, p53 expression was then compared between both groups. RESULTS： In the study, 40 patients were included： 20 with confirmed PHM and 20 with hydropic pregnancy, p53 protein was positive in 60% of patients with PHM and 25% of patients with hydropic pregnancy. The p53 positive rate was significantly higher in patients with PHM （p = 0.027）. Moreover, patients with PHM had a significantly high grade of staining （p 〈 0.001）. CONCLUSION： Our findings indicate that immunohistochemical analysis of p53 protein can be used to distinguish PHM and hydropic pregnancy.