• Clinical and Experimental Reproductive Medicine
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• 제34권3호
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• pp.179-188
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• 2007

목 적: 본 연구에서는 삼핵산 반복서열 확장에 의해 발병하는 헌팅톤병, 척수소뇌성 운동실조증과 X-염색체 취약 증후군 등에 대한 착상전 유전진단을 시행하기 위한 전임상 검사에서 진단 방법을 최적화하는 과정을 통해 얻은 결과들에 대해 기술하고자 한다. 연구방법: 단일 림프구를 이용한 임상전 검사에서는 서로 다른 allele를 갖고 있는 환자의 단일 세포를 사용하였으며, 헌팅톤병과 척수소뇌성 운동실조증에서는 fluorescent semi-nested PCR 시행 후 fragment analysis를 수행하였다. X-염색체 취약 증후군의 경우 multiple displacement amplification (MDA) 방법을 이용한 whole genome amplification에서 얻어진 MDA 산물로 fluorescent PCR을 시도하였다. 결 과: 헌팅톤병의 경우 단일 림프구 시료 모두에서 CAG repeats 증폭에 성공하여 100.0%의 증폭성공률과 14.0% allele drop-out (ADO) rate를, 척수소뇌성 운동실조증의 경우 94.7%의 증폭성공률과 5.6%의 ADO rate을 나타내었다. X-염색체 취약 증후군의 경우 fluorescent semi-nested PCR 방법만으로는 단일 림프구 시료에서 CGG repeats이 증폭되지 않았지만, MDA 산물을 이용한 fluorescent PCR 결과 84.2%의 증폭성공률과 31.3%의 ADO rate을 얻을 수 있었다. 결 론: 본 연구를 통해 헌팅톤병과 척수소뇌성 운동실조증의 착상전 유전진단에는 fluorescent semi-nested PCR 방법의 적용이 가능함을 확인하였으며, X-염색체 취약 증후군의 경우에는 MDA를 이용한 fluorescent PCR 방법을 사용해야 함을 알 수 있었다. 유전자의 변이에 대한 분석이 쉽지 않은 단일 유전자 이상에 대한 착상전 유전진단의 경우 다양한 유전자 분석 방법을 이용한 단일 세포에서의 진단 방법의 최적화 연구가 필수적으로 선행되어야 할 것으로 사료된다.

• Clinical and Experimental Reproductive Medicine
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• 제43권3호
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• pp.169-173
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• 2016

Objective: To determine the preferred regimen for women with adenomyosis undergoing in vitro fertilization (IVF), we compared the IVF outcomes of fresh embryo transfer (ET) cycles with or without gonadotropin-releasing hormone (GnRH) agonist pretreatment and of frozenthawed embryo transfer (FET) cycles following GnRH agonist treatment. Methods: This retrospective study included 241 IVF cycles of women with adenomyosis from January 2006 to January 2012. Fresh ET cycles without (147 cycles, group A) or with (105 cycles, group B) GnRH agonist pretreatment, and FET cycles following GnRH agonist treatment (43 cycles, group C) were compared. Adenomyosis was identified by using transvaginal ultrasound at the initial workup and classified into focal and diffuse types. The IVF outcomes were also subanalyzed according to the adenomyotic region. Results: GnRH agonist pretreatment increased the stimulation duration ($11.5{\pm}2.1days$ vs. $9.9{\pm}2.0days$) and total dose of gonadotropin ($3,421{\pm}1,141IU$ vs. $2,588{\pm}1,192IU$), which resulted in a significantly higher number of retrieved oocytes ($10.0{\pm}8.2$ vs. $7.9{\pm}6.8$, p=0.013) in group B than in group A. Controlled ovarian stimulation for freezing resulted in a significantly higher number of retrieved oocytes ($14.3{\pm}9.2$ vs. $10.0{\pm}8.2$, p=0.022) with a lower dose of gonadotropin ($2,974{\pm}1,112IU$ vs. $3,421{\pm}1,141IU$, p=0.037) in group C than in group B. The clinical pregnancy rate in group C (39.5%) tended to be higher than those in groups B (30.5%) and A (25.2%) but without a significant difference. Conclusion: FET following GnRH agonist pretreatment tended to increase the pregnancy rate in patients with adenomyosis. Further largescale prospective studies are required to confirm this result.

• Clinical and Experimental Reproductive Medicine
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• 제44권2호
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• pp.96-104
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• 2017

Objective: This study aimed to compare the clinical outcomes between in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) in sibling oocytes. Additionally, we evaluated whether the implementation of split insemination contributed to an increase in the number of ICSI procedures. Methods: A total of 571 cycles in 555 couples undergoing split insemination cycles were included in this study. Among them, 512 cycles (89.7%) were a couple's first IVF cycle. The patients were under 40 years of age and at least 10 oocytes were retrieved in all cycles. Sibling oocytes were randomly allocated to IVF or ICSI. Results: Total fertilization failure was significantly more common in IVF cycles than in ICSI cycles (4.0% vs. 1.4%, p<0.05), but the low fertilization rate among retrieved oocytes (as defined by fertilization rates greater than 0% but < 30%) was significantly higher in ICSI cycles than in IVF cycles (17.2% vs. 11.4%, p<0.05). The fertilization rate of ICSI among injected oocytes was significantly higher than for IVF ($72.3%{\pm}24.3%$ vs. $59.2%{\pm}25.9%$, p<0.001), but the fertilization rate among retrieved oocytes was significantly higher in IVF than in ICSI ($59.2%{\pm}25.9%$ vs. $52.1%{\pm}22.5%$, p<0.001). Embryo quality before embryo transfer was not different between IVF and ICSI. Although the sperm parameters were not different between the first cycle and the second cycle, split insemination or ICSI was performed in 18 of the 95 cycles in which a second IVF cycle was performed. Conclusion: The clinical outcomes did not differ between IVF and ICSI in split insemination cycles. Split insemination can decrease the risk of total fertilization failure. However, unnecessary ICSI is carried out in most split insemination cycles and the use of split insemination might make ICSI more common.

• Asian-Australasian Journal of Animal Sciences
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• 제33권12호
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• pp.1922-1929
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• 2020

Objective: The present study aimed to survey seasonal changes in reproductive performance of local cows receiving artificial insemination (AI) in the Pursat province of Cambodia, a tropical country, to investigate if ambient conditions affect the reproductive performance of cows as to better understand the major problems regarding cattle production. Methods: The number of cows receiving AI, resultant number of calving, and calving rate were analyzed for those receiving the first AI from 2016 to 2017. The year was divided into three seasons: cool/dry (from November to February), hot/dry (from March to June), and wet (from July to October), based on the maximal temperature and rainfall in Pursat, to analyze the relationship between ambient conditions and the reproductive performance of cows. Body condition scores (BCS) and feeding schemes were also analyzed in these seasons. Results: The number of cows receiving AI was significantly higher in the cool/dry season than the wet season. The number of calving and calving rate were significantly higher in cows receiving AI in the cool/dry season compared with the hot/dry and wet seasons. The cows showed higher BCSs in the cool/dry season compared to the hot/dry and wet seasons probably due to the seasonal changes in the feeding schemes: these cows grazed on wild grasses in the cool/dry season but fed with a limited amount of grasses and straw in the hot/dry and wet seasons. Conclusion: The present study suggests that the low number of cows receiving AI, low number of calving, and low calving rate could be mainly due to poor body condition as a result of the poor feeding schemes during the hot/dry and wet seasons. The improvement of body condition by the refinement of feeding schemes may contribute to an increase in the reproductive performance in cows during the hot/dry and wet seasons in Cambodia.

• Clinical and Experimental Reproductive Medicine
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• 제45권1호
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• pp.31-37
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• 2018

Objective: To evaluate the pregnancy rate and time to pregnancy after timed coitus with or without superovulation in infertile young women younger than 35 years old with low serum $anti-M{\ddot{u}}llerian$ hormone (AMH) levels ( < 25th percentile). Methods: A total of 202 patients younger than 35 years old were recruited retrospectively between 2010 and 2012. Ninety-eight women had normal serum AMH levels (25-75th percentile), 75 women had low serum AMH levels (5th ${\leq}$ & < 25th percentile) and 29 women had very low serum AMH levels ( < 5th percentile), according to reference values for their age group. Results: The clinical pregnancy rate was positively associated with AMH levels, but this trend did not reach statistical significance (43.9% vs. 41.3% vs. 27.6% in the normal, low, and very low AMH groups, respectively). The time to pregnancy was longer in the very low AMH group than in the normal AMH group ($13.1{\pm}10.9months$ vs. $6.9{\pm}6.1months$, p= 0.030). The cumulative live birth rate over 18 months was lower in the very low AMH group than in the normal AMH group, with marginal significance (20.0% vs. 55.9%, p= 0.051). The duration of infertility was negatively correlated with achieving pregnancy (odds ratio, 0.953; 95% confidence interval, 0.914-0.994; p= 0.026). Conclusion: Conservative management, such as timed coitus with or without superovulation, should be considered in young patients who have low ovarian reserve without any infertility factors. However, for women with a long duration of infertility or very low serum AMH levels, active infertility treatment should be considered.

• 한국가축번식학회지
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• 제25권3호
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• pp.207-217
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• 2001

1999년 5월부터 2000년 5월까지 김제시 관내 32농가 가임암소 836두를 대상으로 번식장애 진단 및 치료를 수행하였다. 번식장애는 직장검사와 초음파 진단기(SA 600, Medison, 5.0MHz rectal linear transducer)를 이용하여 진단한 후 유형별로 분류하였다. 번식장애 치료는 자궁질환우는 자궁세정 또는 질세정을 실시하였고, 개체에 따라 호르몬 처치를 하였다. 난소질환우는 난소질환의 유형에 따라 호르몬 처치를 하고 초음파 소견에 따라 자궁 또는 질세정을 병행하여 실시하였다. 1. 번식장애 대상우 102두중 15두는 임신으로 판정되어 번식장애우는 87두로 가임암소 836두에 대해 10.4%의 번식장애 발생율을 나타내었다. 2. 번식장애는 자궁질환, 난소질환, 자궁과 난소질환 혼합형, 황체존재 무발정형의 4가지로 분류하였다 번식장애우로 분류된 87두중 자궁질환은 33두 37.9%, 난소질환은 30두 34.5%, 자궁과 난소질환 혼합형은 16두로 18.4%, 황체존재 무발정은 8두로 9.2%를 나타내었다. 3 번식장애우 87두중 시험중 번식장애와 함께 병발질환인 요질과 발굽질환이 심하여 번식우로 활용하기가 어렵다고 판단되어 7두를 도태 유도하였다. 4. 번식장애우 87두중에 7두는 도태되었고 치료두수 80두에 대한 치료결과 발정발현우는 74두로 92.5%의 발정발현율을 보였으며, 수태우는 56두로 70.0%의 수태율을 나타내었다. 5. 번식장애 유형별 치료결과는 자궁질환우 32두에 대한 치료후 발정발현우는 31두로 96.8%, 수태우는 23두로 71.9%의 수태율을 나타내었고, 난소질환우는 25두에 대한 치료후 발정발현우는 22두로 55.0%, 수태우는 16두로 64.0%이었으며, 혼합형중 15두에 대하여 발정발현우는 13두로 86.7%, 수태우는 9두로 60.0%의 수태율이었으며, 황체존재 무발정우 8두에 대한 치료후 발정 및 수태 모두 8두에 나타나 발정발현율 및 수태율이 100.0%이었다. 6. 번식장애우의 분만후 경과시기에 따른 발생 여부는 전체 87두중 분만후 0~30일이 2두 2.3%, 30~60일이 6두 6.9%, 60~90일이 12두 13.8%, 90~120일이 18두 20.7%, 120일 이상이 49두로 56.3%로서 분만후 시일이 경과할수록 번식장애우의 발생율이 높은 경향을 보였다. 7. 번식장애우에 번식질환과 함께 병발된 질환을 조사한 결과 번식장애우 87두중 병발질환이 있는 번식장애우는 18두로 20.7%를 나타내었다. 이러한 병발질환을 유형별로 보면 발굽질환우는 5두, 관절질환우는 5두, 뇨질 6두, 직장질루 2두로 나타났다. 병발질환우 18두에 대한 사지질환우는 10두로 55.6%이었다. 8. 번식장애농가 12농가중 7농가가 번식기록 관리를 하지 않았고(58.3%), 또한 5 농가가 운동장이 없거나 활용하지 않아(41.7%) 이러한 요인도 번식장애 발생과 연관이 있다고 판단되었다. 이상의 결과 직장검사에 의존해서 번식장애를 판단하기보다는 초음파 검사와 병행을 할 때 정확한 진단이 이루어질 수 있고 적절한 치료가 이루어져 높은 치료효과를 보일 수 있을 것으로 판단된다. 그리고 번식기록 미비와 운동장의 활용이 적다는 것도 번식장애를 발생시킬 수 있는 사양관리상의 조건이 될 것으로 생각되어 이에 대한 개선방향이 세워져야 할 것으로 판단된다.

• Clinical and Experimental Reproductive Medicine
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• 제24권3호
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• pp.281-292
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• 1997

The objective of this study was to compare retrospectively the survival and pregnancy rates(PR) of cryopresered-thawed embryos obtained from intracytoplasmic sperm injection (ICSI) or conventional in vitro fertilization (IVF). Ninety-six cycles of cryopresered-thawed embryo transfer (ET) were performed in 79 patients from June, 1996 to September, 1997 and grouped as followings: 20 cycles (16 patients) inseminated by ICSI (ICSI Group) and 76 cycles (63 patients) by conventional IVF (IVF Group). Slow-freezing and rapid-thawing protocol was used with 1.5M propanediol (PROH) and 0.1M sucrose as cryoprotectant. All embryos were frozen-thawed at the two pronuclear (2 PN) stage excluding four cycles in which the early cleavage stage embryos were frozen, and allowed to cleave in vitro for one day before ET. The duration from freezing to thawing was comparable in both groups ($mean{\pm}SD$, $112.1{\pm}80.0$ vs. $124.8{\pm}140.1$ days). The age of female ($31.2{\pm}3.4$ vs. $32.6{\pm}3.3$ years) and the endometrial thickness prior to progesterone injection ($9.4{\pm}2.0$ vs. $9.3{\pm}1.8$ mm) were also comparable in both groups. There was no significant difference in the outcomes of cryopreserved-thawed ET between two groups: survival rate ($85.2{\pm}16.1%$ vs. $82.2{\pm}19.7%$), cleavage rate ($96.9{\pm}6.7%$ vs. $94.7{\pm}13.0%$), cumulative embryo score (CES, $54.5{\pm}31.1$ vs. $49.0{\pm}20.0$), preclinical loss rate (5.0% vs. 5.3%), clinical miscarriage rate (0% vs 29.4%), clinical PR per transfer (35.0% vs. 22.4%), implantation rate (9.9% vs. 5.6%), and multifetal PR (42.9% vs. 17.6%). In conclusion, human embryos resulting from ICSI can be cryopreserved-thawed and transferred successfully, and the survival rate and PR are comparable to conventional IVF.

• Clinical and Experimental Reproductive Medicine
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• 제17권1호
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• pp.57-63
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• 1990

The purpose of this study is to improve fertilization rate in IVF-ET program of patients with male infertility used micromanipulation technique, partial zona dissection (PZD) or micro-insemination by sperm transfer (MIST). The results were as follows 1. The fertilization rate of non-micromanipulated oocytes and micromanipulated (PZD) oocytes were 12.5% (n=2) and 42.2% (n=19), respectively, and showed significant differences between two groups (p<0.05). 2. The fertilization rate of micromanipulated (MIST) oocytes was 30% (n=27). 3. The damage rate of Group 1 (PZD) and Group 2 (MIST) were 15.7% (3/19) and 29.6% (8/27), respectively. 4. One pregnancy resulted following replacement of micromanipulated (MIST) embryos in 4 patients.

• Clinical and Experimental Reproductive Medicine
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• 제40권2호
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• pp.90-94
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• 2013

Objective: To evaluate the efficacy of earlier oocyte retrieval in IVF patients with a premature LH surge on hCG day. Methods: One hundred forty IVF patients (164 cycles) with premature LH surge on hCG day were included, retrospectively. We divided them into 2 study groups: LH surge with timed ovum pick-up (OPU) 36 hours after hCG injection (group B, 129 premature cycles), and LH surge with earlier OPU within 36 hours after hCG injection (group C, 35 cycles). Control groups were tubal factor infertility without premature LH surge (group A, 143 cycles). Results: The mean age (year) was statistically higher in group C than in groups A or B ($38.2{\pm}5.4$ vs. $36.2{\pm}4.2$ vs. $36.8{\pm}4.9$, respectively; p=0.012). The serum LH levels (mIU/mL) on hCG day were significantly higher in group B and C than in group A ($22.7{\pm}14.9$ vs. $30.3{\pm}15.9$ vs. $3.2{\pm}2.9$, respectively; p>0.001). Among groups A, B, and C, 4.9%, 31.7%, and 51.4% of the cycles, respectively, had no oocytes, and the overall rates of cycle cancellation (OPU cancellation, no oocyte, or no embryos transferrable) were 15.4%, 65.9%, and 74.3%, respectively. The fertilization rate (%) was significantly higher in group B than in group C ($73.2{\pm}38.9$ vs. $47.8{\pm}42.9$, p=0.024). The clinical pregnancy rate was significantly higher in group C than in groups A and B (44.4% vs. 27.3% vs. 9.1%, respectively, p=0.021). However, the miscarriage rate was also higher in group C than in group B (22% vs. 0%, respectively, p=0.026). Conclusion: Earlier OPU may not be effective in reducing the risk of cycle cancellation in patients with premature LH surge on hCG day. A larger scale study will be required to reveal the effectiveness of earlier ovum retrieval with premature LH surge.

• Clinical and Experimental Reproductive Medicine
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• 제27권1호
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• pp.47-57
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• 2000

Objective: The effects of cryopreservation with or without coculture on the in vitro development of blastomere-biopsied 8-cell mouse embryos were investigated. This experimental study was originally designed for the setup of a preclinical mouse model for the preimplantation genetic diagnosis (PGD) in human. Methods: Eight-cell embryos were obtained after in vitro fertilization (IVF) from F1 hybrid mice (C57BL(표현불가)/CBA(표현불가)). Using micromanipulation, one to four blastomeres were aspirated through a hole made in the zona pellucida by zona drilling (ZD) with acid Tyrode's solution (ATS). A slow-freezing and rapid-thawing protocol with 1.5M dimethyl sulfoxide (DMSO) and 0.1M sucrose as cryoprotectant was used for the cryopreservation of blastomere- biopsied 8-cell mouse embryos. After thawing, embryos were cultured for 110 hours in Ham's F-10 supplemented with 0.4% bovine serum albumin (BSA). In the coculture group, embryos were cultured for 110 hours on the monolayer of Vero cells in the same medium. The blastocyst formation was recorded, and the embryos developed beyond blastocyst stage were stained with 10% Giemsa to count the total number of nuclei in each embryo. Results: The survival rate of embryos after cryopreservation was significantly lower in the blastomere-biopsied (7/8, 6/8, 5/8, and 4/8 embryos) groups than in the non-biopsied, zona intact (ZI) group. Without the coculture, the blastocyst formation rate of embryos after cryopreservation was not significantly different among ZI, the zona drilling only (ZD), and the balstomere-biopsied groups, but it was significantly lower than in the non-cryopreserved control group. The mean number of cells in embryos beyond blastocyst stage was significantly higher in the control group ($50.2{\pm}14.0$) than in 6/8 ($26.5{\pm}6.2$), 5/8 ($25.0{\pm}5.5$), and 4/8 ($17.8{\pm}7.8$) groups. With the coculture using Vero cells, the blastocyst formation rate of embryos after cryopreservation was significantly lower in 5/8 and 4/8 groups, compared with the control, 7/8, and 6/8 groups. The mean number of cells in embryos beyond blastocyst stage was also significantly lower in 4/8 group ($25.9{\pm}10.2$), compared with the control ($50.2{\pm}14.0$), 7/8 ($56.0{\pm}22.2$), and 6/8 ($55.3{\pm}25.5$) groups. Conclusion: After cryopreservation, blastomere-biopsied mouse embryos have a significantly impaired developmental competence in vitro, but this detrimental effect might be prevented by the coculture with Vero cells in 8-cell mouse embryos biopsied one or two blastomeres. Biopsy of mouse embryos after ZD with ATS is a safe and highly efficient preclinical model for PGD of human embryos.