The parental origin correlates with the karyotype of human embryos developing from tripronuclear zygotes / 대한생식의학회지

OBJECTIVE: It has previously been suggested that embryos developing from intracytoplasmic sperm-injected (ICSI) zygotes with three pronuclei (3PN) are endowed with a mechanism for self-correction of triploidy to diploidy. 3PN are also observed in zygotes after conventional in vitro fertilization (IVF). The parental origin, however, differs between the two fertilization methods. Whereas the vast majority of 3PN IVF zygotes are of dispermic origin and thus more likely to have two centrioles, the 3PN ICSI zygotes are digynic in origin and therefore, more likely to have one centriole. In the present study, we examine whether the parental origin of 3PN embryos correlates with the karyotype. METHODS: The karyotype of each nucleus was estimated using four sequential fluorescence in situ hybridizations-each with two probes-resulting in quantitative information of 8 different chromosomes. The karyotypes were then compared and correlated to the parental origin. RESULTS: 3PN ICSI embryos displayed a significantly larger and more coordinated reduction from the assumed initial 3 sets of chromosomes than 3PN IVF embryos. CONCLUSION: The differences in the parental origin-and hence the number of centrioles-between the 3PN IVF and the 3PN ICSI zygotes are likely to be the cause of the differences in karyotypes.

Clinicopathologic features and prognostic factors of malignant phyllodes tumors / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the clinicopathologic features of malignant phyllodes tumors (PT) by histopathologic analyses, immunohistochemical profiling and DNA content assay, and evaluation of the clinical outcome.</p><p><b>METHODS</b>Ten patients with malignant PT from 1999 to 2013 who were treated by surgery were enrolled in this study. The morphologic characteristics were studied under light microscope, standard two-step EnVision method of immunohistochemical staining was used to assess the expression of CK5/6, CKpan, 34β E12, desmin, p63, ER-α, PR, Ki-67, CD34, SMA, p53, p16, bcl-2 and CD117 in the tumors. The corresponding paraffin blocks were also used for flow cytometric DNA content assay. These data were correlated with the follow-up results.</p><p><b>RESULTS</b>The median age of onset was 46.5 years old. The mean tumor size was 7.4 cm (2.0-25.0 cm). At the end of the follow-up period (22 to 125 months), there were tumor recurrences in 3/8 patients and the median time of recurrence was 24 months. Metastasis occurred in 3/8 patients who all died of the tumors. PT had heterogeneous histology, with stromal overgrowth with leaf-like projections, periductal stromal overgrowth, and most commonly, diffuse stromal overgrowth with sarcomatous differentiation. The mean positive index of Ki-67 was 11.4%. The stromal tumor cells were positive for CD34, SMA, p53, p16, and bcl-2 in 3/10, 9/10, 6/10, 8/10, and 4/10 cases, respectively. CD117,ER-α and PR were negative. Interpretable DNA histograms were obtained in nine cases with triploidy in two cases.</p><p><b>CONCLUSIONS</b>The diagnosis of malignant PT should be considered based on the diversity of growth patterns and heterogeneous histology.Ki-67 and CD34 are valuable diagnostic and prognostic factors in patients with malignant PT. Tumors with diffuse stromal overgrowth, heterologous elements, Ki-67 ≥ 20% or aneuploidy are more likely to metastasize.</p>

A Case of Near-triploidy in Myelodysplastic Syndrome with del(5q) Combined with del(1p) and del(13q)

Numerical and structural chromosomal abnormalities are common in hematological malignancies. Near-triploidy (58-80 chromosomes) is a numerical abnormality observed in 3% of adult cases of acute lymphoblastic leukemia. Near-triploidy is rare in myeloid lineage hematologic malignancies and compared to near-triploidy in lymphoid malignancies, near-triploidy in myeloid malignancies is associated with poor outcomes. Few studies on near-triploidy in myelodysplastic syndrome (MDS) have been reported, and the clinicopathologic significance of this condition is still unclear. Here, we report a novel case of MDS with near-triploidy and multiple structural chromosomal abnormalities: del(5q) combined with del(1p) and del(13q). These abnormalities were detected by cytogenetic analysis with array comparative genomic hybridization (CGH). Our results suggest that array CGH can be a useful tool for detecting chromosomal abnormalities in patients with MDS.

Down-regulation of HLA-G attenuates cleavage rate in human triploid embryos

HLA-G is a major histocompatibility complex [MHC], class Ib molecule that is selectively expressed at the fetal maternal interface. It is thought to play a role in protecting the fetus from the maternal immune response. Interestingly, the preimplantation embryo development [Ped] gene product Qa-2 is also a mouse MHC class Ib protein that affects cleavage and division of preimplantation mouse embryos and subsequent embryonic survival. Data from many human in vitro fertilization [IVF] clinics suggest that the mouse Ped phenomenon also exists in human because embryos fertilized at the same time have different cleavage rates and consequently different IVF outcomes. As HLA-G is expressed in human early embryos, it is highly regarded as the functional homologue of Qa-2. Whether HLA-G expression is correlated with the cleavage rate of human embryos has great potential clinical value. In this study, 45 human early abnormal fertilized embryos [3 PN] from patients undergoing in vitro fertilization were used to test the effects of HLA-G knock-down via infection with adenovirus carrying its specific siRNA on the cleavage rate in a 2-day culture period. One-way ANOVA, Post hoc and Chi-square were used to compare groups. A p-value smaller than 0.05 was considered statistically significant. Knocking-down HLA-G in human pre-implantation stage embryos resulted in a higher cell arrest rate and a slower cleavage rate. The results from the present study suggested that HLA-G might play an important role in early human embryo development

Triploidy in a fetus following amniocentesis referred for maternal serum screening test at second trimester.

Amniocentesis was carried out at 17 weeks gestation in a 27-year-old woman, following an abnormal maternal serum screening (MSS) test. MSS test was carried out primarily to estimate the risk of trisomy for chromosome 21. The maternal serum markers used were alpha-fetoprotein (AFP), human chorionic gonadotrophin (hCG), and unconjugated estriol (uE3), together with maternal age. The fetus was identified as screen-positive for Edward’s syndrome (trisomy 18), with low uE3, normal AFP and hCG levels. The calculated risk for trisomy 18 was more than 1:50. To identify any possible chromosomal abnormality, cytogenetic investigation was carried out on the amniotic fluid sample. The fetus’s karyotype showed triploidy with 69, XXX chromosome complement in all the metaphase spreads obtained from three different cultures, using GTG banding technique. Upon termination of the fetus, gross abnormalities indicative of triploidy were present in the fetus.

Roles of p57KIP2 immunohistochemistry and flow cytometry in diagnosis of molar pregnancy / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the value of combined use of paternally imprinted gene product p57(KIP2) immunohistochemistry and flow cytometry in the differential diagnosis of placental hydropic diseases.</p><p><b>METHODS</b>A total of 32 cases of hydropic placenta with DNA polymorphism information were collected, and the genetic results were used as basis for the diagnosis of complete hydatidiform moles (CHM), partial hydatidiform moles (PHM) or hydropic abortions. All cases were examined by histology, p57(KIP2) immunohistochemical staining (EnVision method) and flow cytometry DNA ploidy analysis. The p57(KIP2) immunohistochemical staining and DNA ploidy results were compared with the genetic results.</p><p><b>RESULTS</b>In CHM, p57(KIP2) negative rates were 95.2% (20/21), whereas all the 11 cases of non-CHM (7 cases PHM and 4 cases hydropic abortions) were positive (11/11). In 11 p57(KIP2) -positive cases, 7 cases with triploidy and 4 cases with diploidy by flow cytometry were proven to be PHM and hydropic abortions by genetic analysis, respectively. Overall, 96.9% (31/32) cases of hydropic placentas were correctly diagnosed by combined use of p57(KIP2) immunohistochemistry and flow cytometry.</p><p><b>CONCLUSIONS</b>p57(KIP2) immunohistochemical negativity is a reliable index for the diagnosis of CHM. Combined flow cytometry DNA ploidy and p57(KIP2) immunohistochemistry are useful in the pathological differentiation of CHM, PHM and hydropic abortions.</p>

Severe preeclampsia at 16 weeks' gestation associated with a partial hydatidiform mole and bilateral theca-lutein cysts / 대한산부인과학회지

We present an unusual case in which a patient was diagnosed with the preeclampsia associated with a partial hydatidiform mole and bilateral theca-lutein cysts. The patient newly developed proteinuric hypertension at 16 weeks' gestation. Ultrasound findings showed small multiple cystic spaces in the placenta and a live singleton fetus. Triploid 69, XXX was confirmed at karyotype analysis. Intrauterine fetal death was found at 18 weeks' gestation and termination of pregnancy was performed, and a partial hydatidiform mole was confirmed at pathologic examination. Torsion of bilateral theca-lutein cysts was developed at postpartum one week. Detorsion and aspiration of the bilateral theca-lutein cysts were performed under the laparoscopy.

Cytogenetic Analysis in Spontaneous Abortion / 대한주산의학회잡지

OBJECT: To analyze the frequency and the types of chromosomal abnormalities in conceptus of spontaneous abortion method: 81 cases of products of conception aborted spontaneously were collected From Nov. 1999 to May, 2004 in the cytogenetic laboratory at Kangnam St. Mary's Hospital, Catholic University Medical College. The cytogenetic results,along with clinical information including gestational age at the time of the miscarriage and maternal age, were compiled in a database. RESULTS: Cytogenetic results were obtained from 76 cases (93.8%), of which 33 cases (43.4%) showed chromosome abnormalities. Of 33 chromosomal abnormalities, 12 cases (36.4%) showed monosomy X, including one case of 46,X,+7 and the other case of 44,X, t(13q14q). Autosomal trisomy was also detected in 12 cases (36.4%). The most frequent autosomal trisomy was that of chromosome 18 (4 cases), which occurred exclusively in old maternal age (37.8+/-7.5 years old). Mosaicism was found in 2 cases. Triploidy was identified in 4 cases. Structural abnormalities were identified in 3 cases. CONCLUSION: This study showed that monosomy X and trisomy were the most common chromosomal abnormalities in spontaneous abortion, and autosomal trisomy was dependant on maternal age.

A Case of 21-Monosomy with Holoprosencephaly(Semilobar Type)

Holoprosencephaly of unknown definite causes, has been associated with several chromosome abnormalities involving the autosomes and the sex chromosomes. The most commonly reported associations include dup(3p), del(7q), deletions of chromosome 13, trisomy 13, trisomy 18, and triploidy. In previously reported cases in Korea, none were associated with chromosome 21 anomalies. In conclusion, we reported the first case of holoprosencephaly(semilobar type) associated with pure monosomy 21. We experienced a semilobar type holoprosencephaly with monosomy 21 in a neonate who had multiple congenital anomalies, including an abnormal face, a small thorax with widely spaced hypoplastic nipples and nail hypoplasia, lung hypoplasia with severe scoliosis and cardiac abnormalities. Chromosomal analysis revealed a 45, XY, -21.

A case report of near-triploid in adult acute lymphocytic leukemia / 대한내과학회지

In addition to age, white cell count and immunophenotype, karyotype has been reported to be one of the important prognostic factors in acute lymphocytic leukemias.Furthermore 70 percent of patients with acute B lymphocytic leukemia presented chromosomal abnormalities, which is known to have a close relationship with the prognosis. Among the abnormalities, triploid is rare and known to have the worse prognosis. Structural chromosomal abnormality of the 11q23 band is more common in childhood acute lymphocytic leukemia and has been rarely reported in adult lymphocytic leukemia. We present a case of a 29 year - old male patient with acute lymphocytic leukemia, who had triploid and chromosomal translocation including 11q23 band along with the review of related literature.

Morphohistometric Investigation and bcl-2 Expression in the Placenta of Chromosomally Abnormal Pregnancy

To evaluate the significance of placental histology, a collaborative histological and cytogenetic study was performed on the products of 88 spontaneous abortions, and subsequently bcl-2 immunostaining was performed on 62 cases. The morphometric parameters included were DCIRCLE, FORMSHAPE, CPRATIO, and the expression of bcl-2 immunostainig was graded in four categories (I to IV). The results were as follows: 1) 40% (n=35) were chromosomally abnormal: trisomies predominated (57%, n=20) and was followed by triploidy (14%, n=5), double trisomy (6%, n=2), monosomy X (6%, n=2), inversion (9) (6%, n=2). 2) mean of DCIRCLE in chromosomally abnormal pregnancy was 40 micrometer larger than that in chromosomally normal pregnancy (p=0.012, one side t-test), while no difference was found in FORMSHAPE and CPRATIO between chromosomally abnormal and normal pregnancy. 3) bcl-2 expression was found in syncytiotrophoblast and cytotrophoblast. bcl-2 expression was weaker in chromosomally abnormal pregnancy with intensity I and II of 59% than chromosomally normal pregnancy with intensity I and II of 24%. 4) In comparison bcl-2 expression with DCIRCLE, in chromosomally normal abortion one (10%) in I & II and one (3%) in III & IV showed large DCIRCLE (above 360 micrometer), while 11 (85%) in I & II and 3 (33%) in III & IV in chromosomally abnormal pregnancy. It would mean that bcl-2 protein is necessary in preservation of pregnancy and placental morphology. Abnormal villous diameter and weak bcl-2 expression may be suggestive of chromosomal anomaly. Besides other histologic parameters, application of bcl-2 immunostaining and morphometric analysis probably give more sensitive and specific results in identifying chromosomally abnormal abortion.

The Value of Increased Nuchal Translucency(NT) for the Prediction of Abnormal Pregnancy Outcome / 대한주산의학회잡지

PURPOSE: To evaluate the NT in the first trimester associated with fetal chromosomal abnormality, fetal structural abnormality and adverse pregnancy outcome. METHODS: From Mar. 1993 to Dec. 1997, 133 cases of increased NT were reviewed retrospectively. All these fetues were taken a karyotyping and/or a high resolution ultrasonography for associated anomaly. The NT was measured by transvaginal ultrasonography(TVS) and transabdominal ultrasonography(TAS) if difficulty was encountered with TVS in the period of the 10th-14th week of gestation. The increased NT is approved when it is over 3mm. RESULTS: Of these 133 cases, 102 cases(76.7%) revealed normal outcomes and 31 cases(23.3%) revealed abnormal outcomes such as chromosomal abnormality, structural abnormality and adverse pregnancy outcome, The mean value of NT was 3.44+/-0.68mm in the normal group and 4.93+/-2.70mm in the abnormal group, respectively. The chromosomal abnormalities were found in 16 of the 31 cases: nine Down SD, two Patau syndrome, one Tuner synrome, one triploidy, one 46, XX,i(18q), one 47,iso(Xp)/46,XX one confined placental mosaicism. In 12 cases of the 31 cases, the structural anomalies were detected. Of these 12 cases, 3 cases were eventually developed to cytsic hygroma, 3 cases were cardiac anomaly, 3 cases were renal anomaly, 1 case was congenital diaphragmatic hernia, 1 case was skeletal dysplasia and 1 case was suspected syndrome. The adverse pregnancy outcome such as intrauterine growth restriction, intrauterine fetal death and preterm labor was revealed in 3 cases. In the cases of NT over 7mm, the result was 100% abnormal outcome. CONCLUSION: Increased NT in the first trimester could be a highly efficient the predictor for detection of chromosomal abnormality, structural anomaly and adverse pregnancy outcome.

Cyto - molecular Biologic Characterization of c - myc , erb B and EGF - Receptor in Squamous Cell Carcinoma / 대한피부과학회지

BACKGROUND: Oncogenes and EGF-Receptor(EGFR) may be involved n different stages of the multistep carcinogenesis process. A specific pattern of karyotypic abnormalities in solid tumors can be detected by cytogenetic methods. OBJECTIVE: This study is intnded to observe the cytomolecular kiologic chracterization of c-myc, erb B and EGFR genes in squasnous cell carcinoma(SCC) of the skin and cervix. METHODS: We have eytogenet,ically examined the short-term culturs from SCC. The rearrangement, amplification or expressi.on of erb B, c-myc, and EGFR genes were studied by Southern blot, analysis of genomic DNA and by slot blot analysis of tota! RNA extracted from biopsies of normal skin and SCC tissues. EGFR expression was examined immunohistochemially using monoclonal antibodies and the localizat,ion of the c-myc oncogene mRNA by in situ hybridization. RESULTS: A remarkably structural aberration was del 6(q21-qter) counted 20 metaphases among 28 metaphases ana1yzed. In nunierical aberration, all chromosomes were lost or gained randomly. Amenploid including triploid and tetraploid were observed in 8 metaphases, 6 tumor cells contained marker chromosome. In Southern blot analysis, rearrangement and amglificaton of EGFR in primary squamous cell carcinoma of cervix uteri and skin respectively. In slot blot analysis, the levels of c-myc, erb B and EGFR mRNA increaaed respectively 3.5, 2.5 and 2.8 times in SCC when compared to normal tissues. In immunoperoxidase stain, EGFR was present, in SCC where keratinocytes with strong cyto-plasmic staining but no membr, line labelling, where as in normal skin the were primarily present in t,he membrane and cytoplasm of basal cells. In situ hybridization with c-myc cDNAs allowed detection of grains representative of biotin labelled cDNA-mRNA hybrids in the frozden section of SCC tissues. CONCLUSION: These results suggest that specific patterns of karyotypir abnormalites, rearrangement, or amplification of EGFR gene, and overexpression of oncogenes and EGFR gene may be associated with the carcinogenesis of SCC.

Chromosomal Changes in Transitional Cell Carcinoma of the Bladder (Preliminary Report) / 대한비뇨기과학회지

3 cases of transitional cell carcinoma were subjected to detailed cytogenetic analysis. All three were superficial (T1G I , T1G III, T1G I). Case 1 and case 3 (all T1G I) had diploidy modal chromosomal number but case 2 (TtGIII) had partly triploidy and tetraploidy chromosome. Case 3 showed marker chromosomes and in case 2 and 3, breakage of long arm of the second chromosome was seen.