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Effect of Ethylene Glycol (EG) and 1,2-Propanediol (PROH) on the Survival and the Development of Mouse and Human Embryos after Slow Freezing/Rapid Thawing Protocol (생쥐와 인간배아의 완만동결-급속융해 후 생존률과 배아발생에 미치는 Ethylene Glycol (EG)과 1,2-Propanediol (PROH)의 영향)

  • Kim, Tae-Hyung;Cha, Soo-Kyung;Lee, Dong-Ryul;Han, Jee-Eun;Lee, Woo-Sik;Yoon, Tai-Ki;Cha, Kwang-Yul;Chung, Hyung-Min
    • Clinical and Experimental Reproductive Medicine
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    • v.31 no.1
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    • pp.9-17
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    • 2004
  • Objective: The aim of this study were to compare the effects of EG and PROH on cryopreservation of mouse and human embryos, and to find the optimal protocol for embryo freezing. Methods: Human embryos derived from fertilized eggs showing 3 pronuclei (PN) and mouse embryos were divided into two groups respectively: dehydrated with 1.5 M EG + 0.2 M sucrose or 1.5 M PROH + 0.2 M sucrose using the slow freezing method. Moreover mouse embryos were controlled the exposure time of cryoprotectant during dehydration or rehydration steps. Results: The survival rates of human embryos were 79.2% (84/106) in EG group and 77.9% (88/113) in PROH group. In mouse embryos, the survival and development rates up to blastocyst were 70.6% (245/347), 44.1% (123/279) in EG group and 62.1% (198/319), 45.1% (123/279) in PROH group, respectively. However, in EG group, partially damaged embryos after thawing were decreased compared to PROH group. In combination group, when the exposure time during dehydration and rehydration were reduced, the survival and embryonic developments were increased slightly, but not significant. Conclusion: Cryopreservation of mouse and human embryos at cleavage stage by using EG or PROH exhibited no statistical difference in the survival rate and/or developmental rate to blastocyst. However, the use of EG for cryopreservation of embryos might reduce the exposure time of the cryoprotectant because of a high permeation of EG and result in lessen its toxic effects.

Pseudoisodicentric X chromosome in a female with primary amenorrhea (원발성 무월경 여성에서 관찰된 Pseudoisodicentric X 염색체)

  • Park, Sang-Hee;Shim, Sung-Han;Chin, Mi-Uk;Kang, Su-Jin;Bae, Sung-Mi;Sohn, Soo-Min;Cha, Dong-Hyun;Yoon, Tae-Ki;Cho, Jung-Hyun
    • Journal of Genetic Medicine
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    • v.5 no.1
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    • pp.61-64
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    • 2008
  • A 24-year-old female with primary amenorrhea was referred for a chromosome study. The karyotype of the patient was 46,X,der(X) under initial GTG-banding analysis. Fluorescence in situ hybridization (FISH) analysis with an LSI Kallmann (KAL) region probe [probes for Xp22.3(KAL) and CEP(X) for control] was carried out. The abnormal chromosome was KAL- and CEP(X)${\times}2$. In addition, interphase FISH analysis revealed the patient to be mosaic for two different cell lines: 90% of cells had three signals and 10% of the cells had only one signal for CEP(X). Based on these results, the karyotype of the patient was 45,X/46,X,psu idic(X)(p22.1), which is partial trisomy for Xqter${\rightarrow}$Xp22.1 and partial monosomy for Xpter${\rightarrow}$Xp22.1. This karyotype was considered a variant of Turner syndrome. In summary, Idic(X) and low-level mosaicism was successfully characterized by FISH analysis with a CEP(X) probe.

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Analysis of the Impact on Community Health after Accidental Leak of Hydrofluoric Acid (일개 응급의료센터에서 분석한 불화수소산 대량 누출이 지역사회 건강에 미친 영향)

  • Kim, Young Gab;Lee, Ju Taek;Park, Sang Hyun;Lee, Chan Hee;Choe, Michael Sung Pil;Je, Dong Wook;Lee, Chang Jae;Ko, Taei;Jo, Hye Jung
    • Journal of The Korean Society of Clinical Toxicology
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    • v.11 no.2
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    • pp.106-113
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    • 2013
  • Purpose: The purpose of this study is to verify the influence of a massive hydrofluoric acid spill on community health through patients who claimed to have been exposed. Methods: We analyzed 2459 patients who visited our emergency department with the claim of exposure to hydrofluoric acid, and retrospective analyses were performed. We analyzed changes in numbers of visitors per day from the day of the accidental hydrofluoric acid spill, symptoms presented by the 1924 patients, and general characteristics. Comparisons of symptoms and hematologic characteristics were made between the initially set evacuation zone(1.3 km radius parameters from the spill) and the outer zone. Results: A total of 2,459 patients who claimed exposure visited our ED from 27 September 2012 to 23 October 2012, and there was a significant increase in the number of visiting patients from day 8 of the hydrofluoric acid spill. The most common complaints were a sore throat, 729(37.9%) and no specific symptom with health concern, 547 (28.4%). Statistically significant findings were pulmonary symptoms (p=0.001), nasal symptoms (p=0.001), diarrhea (p=0.023), and skin symptoms (p=0.007). In hematologic study, a statistically significant difference was observed in white blood cell count (p=0.018), creatine phosphokinase (p<0.001), erythrocyte sediment rate (p=0.013), and phosphorus (p<0.001). Conclusion: A significant increase in the number of patients was observed one week after the accidental spill of hydrofluoric acid. The most frequent symptoms were sore throat, headache, cough, and sputum. Statistically significant increase in creatine phosphokinase level and decrease in phosphorus level were noted in patients within the evacuation zone.

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Xenografting of the Human Vitrified Ovarian Tissues into the Immune Deficient Animal (사람 난소조직의 초자화 냉동보존과 면역결핍 동물에의 이식)

  • Lee, Kyung-Ah;Yoon, Se-Jin;Lee, Sook-Hyun;Shin, Chang-Sook;Choi, An-Na;Cho, Yong-Seon;Yoon, Tae-Ki;Cha, Kwang-Yul
    • Clinical and Experimental Reproductive Medicine
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    • v.27 no.2
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    • pp.145-149
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    • 2000
  • Objective: The present study was conducted to evaluate the viability of germ cells from the adult and fetal ovarian tissues after vitrification followed by xenografting. Method: The human adult ovarian tissues were obtained from 33 years old patient, and the fetal ovarian tissues were obtained from 22 weeks and 25 weeks in gestation. Ovarian tissues were cryopreserved by vitrification with 5.5 M ethylene glycol (EG 5.5) and 1.0 M sucrose as cryoprotectants. Adult and fetal ovarian tissues were pre-equilibrated with EG 5.5 at room temperature for 10 and 5 minutes, respectively and plunged into liquid nitrogen immediately. Frozen-thawed tissues were xenografted into NOD-SCID mice to evaluate the viability and capacity for further growth of the primordial follicles. Grafts were recovered from the recipients 4 weeks after transplantation and histological analysis was accomplished. Result and Conclusion: Grafts recovered 4 weeks after transplantation contained less number of oocytes and primordial follicles compared to that of the fresh tissues. Survived follicles were mainly primordial and intermediary with larger diameter and more granulosa cells. It is confirmed that 1) the ovarian tissues were healthy and the germ cells were survived after vitrification, and 2) the survived fetal primordial follicles after vitrification resumed the growth in the xenografts.

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Pinopode Development 2-days after Oocyte Retrieval in the Human IVF Patients (체외수정 환자에서 난자회수 2일째의 자궁 내막의 Pinopode의 발달)

  • Lee, Kyung-Ah;Han, Sei-Yul;Choi, Dong-Hee;Lee, Woo-Sik;Yoon, Tae-Ki;Cha, Kwang-Yul
    • Clinical and Experimental Reproductive Medicine
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    • v.25 no.1
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    • pp.51-58
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    • 1998
  • 본 연구는 체외수정 프로그램에 참여하는 환자에 있어서 난자회수 이틀째의 자궁내막의 발달상태를 알아보기 위하여 pinopode의 발달상태, 에스트로젠 및 프로제스테론 수용체의 발현을 관찰하였다. 생검한 자궁내 막 조직 을 양분하여, 절반은 전사전자 현미경 (scanning electron microscope)으로 pinopode를 관찰하기 위하여 2.5% glutaraldehyde와 2% paraformaldehyde로 고정하였고, 나머지 절반은 dating 및 스테로이드 수용체의 면역조직화학적 측정 (immunocytochemistry)을 위하여 10% formalin으로 고정하였다. 모두 12명의 환자중 8명에서 pinopode가 관찰되었으며, pinopode 발달이 관찰되지 않은 환자들은 hCG 주사를 맞는 날의 estradiol (E2)의 혈중농도가 600 pg/mL이하로 낮았다. 본 연구의 결과로부터 자궁내막의 발달상태를 알아보기 위해서는 지금까지 일반적으로 사용되어 오던 dating이나 스테로이드 수용체의 면역조직화학적 측정법 이외에도 pinopode를 관찰함으로써 조금 더 정확한 진단을 할 수 있으리라고 사료되며, pinopode의 발달은 E2의 혈중농도와 관계가 있을 것으로 추정된다.

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Eupatilin treatment inhibits transforming growth factor beta-induced endometrial fibrosis in vitro

  • Lee, Chang-Jin;Hong, Seon-Hwa;Yoon, Min-Ji;Lee, Kyung-Ah;Choi, Dong Hee;Kwon, Hwang;Ko, Jung-Jae;Koo, Hwa Seon;Kang, Youn-Jung
    • Clinical and Experimental Reproductive Medicine
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    • v.47 no.2
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    • pp.108-113
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    • 2020
  • Objective: Endometrial fibrosis, the primary pathological feature of intrauterine adhesion, may lead to disruption of endometrial tissue structure, menstrual abnormalities, infertility, and recurrent pregnancy loss. At present, no ideal therapeutic strategy exists for this fibrotic disease. Eupatilin, a major pharmacologically active flavone from Artemisia, has been previously reported to act as a potent inducer of dedifferentiation of fibrotic tissue in the liver and lung. However, the effects of eupatilin on endometrial fibrosis have not yet been investigated. In this study, we present the first report on the impact of eupatilin treatment on transforming growth factor beta (TGF-β)-induced endometrial fibrosis. Methods: The efficacy of eupatilin on TGF-β-induced endometrial fibrosis was assessed by examining changes in morphology and the expression levels of fibrosis markers using immunofluorescence staining and quantitative real-time reverse-transcription polymerase chain reaction. Results: Eupatilin treatment significantly reduced the fibrotic activity of TGF-β-induced endometrial fibrosis in Ishikawa cells, which displayed more circular shapes and formed more colonies. Additionally, the effects of eupatilin on fibrotic markers including alpha-smooth muscle actin, hypoxia-inducible factor 1 alpha, collagen type I alpha 1 chain, and matrix metalloproteinase-2, were evaluated in TGF-β-induced endometrial fibrosis. The expression of these markers was highly upregulated by TGF-β pretreatment and recovered to the levels of control cells in response to eupatilin treatment. Conclusion: Our findings suggest that suppression of TGF-β-induced signaling by eupatilin might be an effective therapeutic strategy for the treatment of endometrial fibrosis.

Screening for down syndrome using trophoblast retrieval and isolation of the cervix: preliminary study

  • Lee, Min Jin;Kim, Soo Hyun;Park, Hee Jin;Shim, Sung Han;Jang, Hee Yeon;Cha, Dong Hyun
    • Journal of Genetic Medicine
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    • v.17 no.2
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    • pp.68-72
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    • 2020
  • Purpose: Trisomy 21, the cause of Down syndrome (DS) with various medical problems, is the most common aneuploidy during the fetal period. For diagnosis, a non-invasive screening test using maternal blood, which cannot be confirmed and invasive confirmation test with a risk of miscarriage, may be performed. The trophoblast retrieval and isolation of the cervix (TRIC) have been proposed by some researchers as an alternative to overcome the limitations of current tests. We experimented using TRIC to identify the possibility of trisomy 21 for the first time in Asia. Materials and Methods: Three cases of DS were analyzed confirmed by invasive tests (chorionic villus sampling, amniocentesis). All samples of trophoblasts immediately were immersed in phosphate-buffered saline and processed with formalin for fixation. The trophoblasts were isolated using an anti-human leukocyte antigen-G antibody coupled to magnetic nanoparticles. β-human chorionic gonadotropin (hCG)-expressing cells were considered as trophoblast cells, and the detection rate calculated. DS was confirmed by fluorescence in situ hybridization (FISH). Results: The mean trophoblast detection rate using β-hCG was 78.1%, and the detection rate using FISH was 22.2%. In all cases, the trisomy of chromosome 21 was identified. Conclusion: Trophoblast can be obtained from the five weeks of gestation and has a high detection rate, so it is noted that it can replace the current prenatal genetic test. To realize the clinical application as a prenatal genetic test, we will need additional efforts to identify trisomy 21 as well as other chromosomal abnormalities in future large-scale studies.

Identification of unbalanced complex chromosomal rearrangements in IVF-derived embryos during NGS analysis of preimplantation genetic testing: A case report

  • Yu, Eun Jeong;Kim, Min Jee;Park, Eun A;Hong, Ye Seul;Park, Sun Ok;Park, Sang-Hee;Lee, Yu Bin;Yoon, Tae Ki;Kang, Inn Soo
    • Journal of Genetic Medicine
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    • v.19 no.1
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    • pp.14-21
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    • 2022
  • Complex chromosome rearrangements (CCRs) are structural chromosomal rearrangements involving at least three chromosomes and more than two breakpoints. CCR carriers are generally phenotypically normal but related to higher risk of recurrent miscarriage and having abnormal offspring with congenital anomalies. However, most of CCR carriers are not aware of their condition until genetic analysis of either abortus or affected baby or parental karyotyping is performed. Herein, we present the case that CCR carrier patients can be identified by preimplantation genetic testing of preimplantation embryos. An infertile male patient with severe oligoasthenoteratozoospermia was diagnosed balanced reciprocal translocation, 46,XY,t(3;11) (p26;p14) at first. After attempting the first preimplantation genetic testing for structural rearrangement (PGT-SR) cycle, we found the recurrent segmental gain or loss on 21q21.3-q22.3 of five out of nine embryos. As a result of karyotype re-analysis, the patient's karyotype showed a balanced CCR involving chromosomes 3, 11, and 21 with three breakpoints 3p26, 11p14, and 21q21. The patient underwent two PGT-SR cycles, and a pregnancy was established after the transfer of an euploid embryo in the second cycle. Amniocentesis confirmed that the baby carried normal karyotype without mosaicism. At 37 weeks gestation, a healthy girl weighting 3,050 g was born.