OBJECTIVE: To evaluate the impact of endometriosis on IVF-ET cycles and to compare IVF outcomes between stage I/II and stage III/IV endometriosis. METHODS: We analyzed 697 patients (1,199 cycles) with endometriosis (stage I-II: 638 cycles, stage III-IV: 561 cycles) and 325 pts (459 cycles) with tubal factor as controls between January 1994 and April 2004. Pts with endometriosis were diagnosed by laparoscopy and medical and surgical treatment were done in 353 cycles (55.3%) and 466 cycles (83.1%) of stage I-II/stage III-IV endometriosis. Cycles with age>35 years or FSH>20 mIU/mL or severe male factor infertility were excluded. RESULTS: The number of retrieved oocytes (9.97+/-7.2 vs. 13.4+/-7.9 (p<0.0001)), total number of embryos (6.5+/-4.8 vs. 9.1+/-5.6 (p<0.0001)), and good quality embryos (2.43+/-1.6 vs. 2.74+/-1.7 (p=0.013)) significantly decreased in stage III-IV endometriosis than in control. But pregnancy rate of stage III-IV endometriosis was comparable with control (35.7% vs. 36.8%). Fertilization rate and number of total embryos were lower in stage I-II endometriosis than in control (64.8+/-22.9 vs. 70.8+/-20.8 (p<0.0001), 7.6+/-5.0 vs. 9.1+/-5.6 (p<0.0001)). In patients with medical and surgical treatment of endometriosis, pregnancy rate and live birth rate was significantly lower in stage I-II than in stage III-IV endometriosis (29.2 vs. 36.2 (%), p=0.045, 23.9 vs. 31.5 (%), p=0.043). There was no difference in the mean age, but the duration of infertility was significantly longer (56.5+/-26.3 vs. 46.9+/-25.8 (mon), p<0.0001) and fertilization rate was lower (64.7+/-23.3 vs. 70.5+/-22.7 (%), p=0.001) in stage I-II than stage III-IV endometriosis. CONCLUSION: We suggest that IVF should be considered earlier in patients with minimal to mild endometriosis because of significantly decreased fertilization rates.
Embryonic Structures ; Endometriosis* ; Female ; Fertilization ; Humans ; Infertility ; Laparoscopy ; Live Birth ; Male ; Oocytes ; Pregnancy Rate

No abstract available.

Recently, through the development of the primer extension preamplification(PEP) method which amplifies the whole genome, simultaneous multiple DNA analysis has become possible. Whole genome from each single cell can be amplified using 15 base oligonucleotide random primer. The greatest advantage of PEP-PCR is the ability to investigate several loci simultaneously and confirm results by analysing multiple aliquots for each locus. This technique led to the development of preimplantation genetic disease diagnosis using blastomere from early embryo, sperm, polar body and oocyte. In this study, we applied PEP-PCR in 20 cases of single amniocyte and 20 cases of single chorionic villus cell for the clinical application of the prenatal and preimplantational genetic diagnosis. We analysed 7 gene loci simultaneously which are 46, 47 exons related to dystrophin gene, two VNTR (variable number tandem repeat) markers using 5'toysIII, 3'CA related to dystrophin gene and DYZ1, DYZ3, DYS14 regions on chromosome Y. In all the tests, 97.5% of PEP-PCR amplifications with single cells were successful. We obtained 38/40 (95%) accuracy in gender determination through chromosome analysis comparison. Therefore, these results have significant implications for a sperm or oocyte analysis and prenatal or preimplantational genetic diagnosis.
Blastomeres ; Chorionic Villi ; Diagnosis ; DNA ; Dystrophin* ; Embryonic Structures ; Exons ; Genome ; Oocytes ; Polar Bodies ; Spermatozoa

OBJECTIVE: To evaluate the viability and the characteristics of shed endometrial tissues obtained from menstrual fluid during in-vitro culture. METHODS: The menstrual fluids were collected using Wallace catheter from uterine cavity in 10 women with regular menstruation. The menstrual fluids were washed twice, and the pellets, containing blood cells and shed endometrium, were collected and diluted fivefold with Ham's F-10 medium containing 10% fetal bovine serum. The cell suspension was placed on culture dishes, and cultured for 7 days in an incubator. To evaluate the characteristics of the cultured endometrial cells, immunohistochemical (IHC) staining was performed using anti-cytokeratin and anti-vimentin antibody. RESULTS: The mean volume of menstrual fluids and pellets were 0.7ml and 0.3ml, respectively. Only 15% of the shed endometrial tissues were attached and proliferated in culture dishes, which was considered to have viability. Initially, endometrial epithelial cells and fibroblasts were attached and proliferated, and the area of these cells was increased according to prolong the culture time. Stromal cell colonys were located and proliferated on the epithelial cells. IHC staining showed strongly positive for cytokeratin in epithelial cells and for vimentin in stromal cells. In the confocal microscopic observation of 3-dimensional structure of cultured endometrium, cytokeratin-positive cells (epithelial cells) were located in the pheriphery and cytokeratin-negative cells (stromal cells) inside of the structure. CONCLUSION: From our study, shed endometrial tissues in menstrual fluid showed meaningful viability and closed relationship between epithelial cells and stromal cells during in-vitro culture. Thus, we suggest that the in-vitro culture system of shed endometrium is a suitable model for researches of endometriosis.
Blood Cells ; Catheters ; Endometriosis ; Endometrium ; Epithelial Cells ; Female ; Fibroblasts ; Humans ; Incubators ; Keratins ; Menstruation ; Stromal Cells ; Vimentin

OBJECTIVE: To investigate the association between endometriosis and polymorphisms of N-acetyl transferase 2 (NAT2), glutathione S-transferase M1 (GSTM1), and cytochrome P450 (CYP) 1A1 genes in Korean infertile patients. MATERIALS AND METHODS: A total of 303 infertile patients who had undertaken diagnostic laparoscopy during January, 2001 through December, 2003 at Samsung Cheil Hospital enrolled in this study. The patients were grouped according to laparoscopic findings: minimal to mild endometriosis (group I: n=147), moderate to severe endometriosis (group II: n=57), normal pelvic cavity (n=99). Peripheral blood was obtained and genomic DNA was extracted. The genotypes of each genes were analyzed using polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP). For NAT2, RFLP was used to detect the wild type (wt) and mutant (mt) alleles, enabling classification into slow (mt/mt) or fast (wt/wt or wt/mt) acetylation genotypes. For GSTM1, PCR was used to distinguish active (+/- or +/+) from null (-/-) genotypes. For CYP1A1, MspI digestion was used to detect the wild type (A1A1), heterozygote (A1A2) or mutant (A2A2) genotypes. RESULTS: The genotype frequencies of NAT2 slow acetylator was 12.8%, 10.9%, 12.8% in group I, group II and control, respectively. The genotype frequencies of GSTM1 null mutation was 55.3%, 41.8%, 53.2% in group I, group II and control, respectively. The genotype frequencies of CYP1A1 MspI polymorphism was 16.3%, 9.1%, 18.1% in group I, group II and control, respectively. No significant difference was observed between endometriosis and normal controls in the genotype frequencies of the NAT2, GSTM1, CYP1A1 MspI polymorphism. CONCLUSION: The NAT2, GSTM1, CYP1A1 gene polymorphism may not be associated with the susceptibility of endometriosis in Korean women.
Acetylation ; Alleles ; Classification ; Cytochrome P-450 CYP1A1 ; Cytochrome P-450 Enzyme System* ; Cytochromes* ; Digestion ; DNA ; Endometriosis* ; Female ; Genotype ; Glutathione Transferase* ; Glutathione* ; Heterozygote ; Humans ; Laparoscopy ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Transferases*

OBJECTIVES: We have previous reported that thawed testicular sperm and sperm extracted from seminiferous tubule could achieved optimal fertilization and pregnancy in azoospermic patients. However, thawed testicular sperm did not show motility in many cases. Therefore we studied viability of immotile sperm extracted from frozen-thawed seminiferous tubule using hypo-osmotic swelling (HOS) test and eosin-Y test. MATERIALS AND METHODS: After sperm extraction using for ICSI, the remained sections of seminiferous tubules were frozen with computerized freezer. For thawing and preparation of testicular sperm, the seminiferous tubules were thawed by removing from LN2 and letting them at room temperature for 10 min followed by 37degrees C water bath for 10 min. The prepared samples were washed for free of preservation medium and sperm preparation method described previous. Sperm was suspended in 0.1 ml hypoosmotic solution. After 30 minutes, the type of distally coiled sperm were assessed. RESULTS: In 44 cases of cryopreservation of seminiferous tubules in obstructive azoospermic patients, the fertilization rates with 2PN were 71.4% and pregnancy rates were 34.1%. The presence of motile spermatozoa on subsequent post-thaw testicular sperm remarked 15.1% and were increased to 77.3% just before ICSI. After sperm extracted from frozen-thawed seminiferous tubule, 3 hrs later in in vitro culture, the cases of presence of motile sperm, reaction of hypo-osmotic swelling test and viable sperm were 63.6% (28/44), 93.2% (41/44), and 77.3% (34/44), respectively. CONCLUSIONS: Just after post-thawed testicular sperm did not showed motility. Although motility was gained after in vitro culture, many cases showed non-motile sperm until optimal insemination time. However, HOS test showed positive reaction in non-motile sperm. Therefore, HOS test is an alternative method for the selection of viable sperm for ICSI.
Baths ; Cryopreservation ; Fertilization ; Humans ; Insemination ; Linear Energy Transfer ; Pregnancy ; Pregnancy Rate ; Seminiferous Tubules ; Sperm Injections, Intracytoplasmic ; Spermatozoa* ; Water

OBJECTIVES: Pelvic tuberculosis (TB) causes infertility despite of anti-TB chemotherapy and IVF-ET is effective treatment to achieve pregnancy. The aim of this study is to assess the outcomes of IVF-ET in pelvic TB, especially according to main Tb lesion, and to investigate the factors affecting the successful outcome. METHODS: A total of 135 IVF-ET cycles were performed in 54 patients with pelvic TB and the outcome was compared with that of control group with tubal factor not associated with TB in 301 cycles, 227 patients. Anti-TB chemotherapy was performed in the patients with pelvic TB. Pregnancy rate was compared according to main TB lesion as salpingitis, peritonitis, and endometritis. In the patients with endometrial TB, when complicated with uterine synechia, hysteroscopic lysis was done before IVF-ET and pregnancy rate was compared according to the presence of uterine synechia. RESULTS: There was no significant difference in peak E2 (2,790+/-280.1 vs 2,554+/-101.2, p>0.05), the number of retrieved oocytes (13.5+/-0.7 vs 12.5+/-0.4, p>0.05) and fertilized oocytes (7.7+/-0.5 vs 7.8+/-0.3, p>0.05) between patient and control group. Clinical pregnancy rate per transfer in pelvic TB group was 22.9% and showed no difference from that of control group (24.3%, p>0.05). Although it was not statistically significant, pregnancy rate in the endometrial Tb (18%) was lower than that in the salpingitis (28.5%) or peritonitis (26.5%) (p>0.05). In the endometrial TB with uterine synechia, pregnancy rate was significantly lower than that of the patients without synechia even after hysteroscopic lysis (9.7% vs 31.6%, p<0.05). CONCLUSION: IVF-ET after anti-TB chemotherapy is the most effective treatment to achieve pregnancy in infertile patients with pelvic TB. Because the presence of endometrial TB and resulting uterine synechia affects the outcome of IVF-ET, thorough evaluation for endometrium with endometrial biopsy and hysteroscopy is important to predict the prognosis of IVF-ET treatment.
Biopsy ; Drug Therapy ; Embryo Transfer* ; Embryonic Structures* ; Endometritis ; Endometrium ; Female ; Fertilization in Vitro* ; Humans ; Hysteroscopy ; Infertility ; Oocytes ; Peritonitis ; Pregnancy ; Pregnancy Rate ; Prognosis ; Salpingitis ; Tuberculosis*

OBJECTIVES: Pelvic tuberculosis (TB) causes infertility despite of anti-TB chemotherapy and IVF-ET is effective treatment to achieve pregnancy. The aim of this study is to assess the outcomes of IVF-ET in pelvic TB, especially according to main Tb lesion, and to investigate the factors affecting the successful outcome. METHODS: A total of 135 IVF-ET cycles were performed in 54 patients with pelvic TB and the outcome was compared with that of control group with tubal factor not associated with TB in 301 cycles, 227 patients. Anti-TB chemotherapy was performed in the patients with pelvic TB. Pregnancy rate was compared according to main TB lesion as salpingitis, peritonitis, and endometritis. In the patients with endometrial TB, when complicated with uterine synechia, hysteroscopic lysis was done before IVF-ET and pregnancy rate was compared according to the presence of uterine synechia. RESULTS: There was no significant difference in peak E2 (2,790+/-280.1 vs 2,554+/-101.2, p>0.05), the number of retrieved oocytes (13.5+/-0.7 vs 12.5+/-0.4, p>0.05) and fertilized oocytes (7.7+/-0.5 vs 7.8+/-0.3, p>0.05) between patient and control group. Clinical pregnancy rate per transfer in pelvic TB group was 22.9% and showed no difference from that of control group (24.3%, p>0.05). Although it was not statistically significant, pregnancy rate in the endometrial Tb (18%) was lower than that in the salpingitis (28.5%) or peritonitis (26.5%) (p>0.05). In the endometrial TB with uterine synechia, pregnancy rate was significantly lower than that of the patients without synechia even after hysteroscopic lysis (9.7% vs 31.6%, p<0.05). CONCLUSION: IVF-ET after anti-TB chemotherapy is the most effective treatment to achieve pregnancy in infertile patients with pelvic TB. Because the presence of endometrial TB and resulting uterine synechia affects the outcome of IVF-ET, thorough evaluation for endometrium with endometrial biopsy and hysteroscopy is important to predict the prognosis of IVF-ET treatment.
Biopsy ; Drug Therapy ; Embryo Transfer* ; Embryonic Structures* ; Endometritis ; Endometrium ; Female ; Fertilization in Vitro* ; Humans ; Hysteroscopy ; Infertility ; Oocytes ; Peritonitis ; Pregnancy ; Pregnancy Rate ; Prognosis ; Salpingitis ; Tuberculosis*

OBJECTIVES: The most common chromosomal abnormality contributing to recurrent abortion is the balanced chromosomal translocation. However the exact incidence of fetal losses are still unknown. The objectives of this study were to evaluate the incidence of fetal chromosomal abnormalities and outcome of pregnancy in recurrent miscarriage couples with balanced translocation. DESIGN: A retrospective analysis of recurrent spontaneous abortion patients with balanced chromosomal translocation. MATERIALS AND METHODS: Cytogenetic analysis was performed in 56 couples with history of recurrent abortions from 1995 to 1999. The use of high resolution banding technique and fluorescent in situ hybridization (FISH) in the chromosomal analysis has made the precise evaluation of chromosome aberrations. RESULTS: Among 56 couples, 42 patients had reciprocal translocation and 14 had Robertsonian translocation. Chromosomal aberrations were more frequent in women (36 cases) than in men (20 cases). Prenatal cytogenetic analyses were carried out in 14 subsequent pregnancies for carrier couples with balanced translocation. The fetal karyotypes showed that 5 cases (35.7%) was normal, 8 (57.1%) were balanced translocation, and 1 (7.1%) was unbalanced translocations. And cytogenetic analyses were done on 15 subsequent chorionic villi samples of abortuses for carrier couples with balanced translocations. Fourteen of fifteen abortuses (93.3%) were abnormal karyotype. CONCLUSIONS: Although the incidence of chromosomal imbalance in the fetuses was relatively low in prenatal cytogenetic analysis, individuals with balanced translocations are predisposed to giving birth to malformed offsprings with chromosomal imbalance (partial trisomy or monosomy). Therefore we recommend preimplantation genetic diagnosis (PGD) for recurrent abortions with balanced translocation and preventing the birth of offspring with chromosomal abnormalities.
Abnormal Karyotype ; Abortion, Habitual* ; Abortion, Spontaneous ; Chorionic Villi ; Chromosome Aberrations* ; Cytogenetic Analysis ; Family Characteristics* ; Female ; Fetus ; Humans ; In Situ Hybridization, Fluorescence ; Incidence* ; Karyotype ; Male ; Parturition ; Pregnancy ; Preimplantation Diagnosis ; Retrospective Studies ; Translocation, Genetic ; Trisomy

Preimplantation genetic diagnosis (PGD) provides practical option to prevent the termination of pregnancy and miscarriage in couples with high risk of genetic disease or recurrent spontaneous abortion. In balanced chromosomal translocation, PGD can reduce the abortion rate and with PGD for aneuploidy screening, higher implantation rate and lower abortion rate can be obtained in patients with poor reproductive prognosis. Therefore PGD is widely used in ART for improving IVF efficiency. With technical development in single cell, such as FISH, PCR, CGH and microarray, the indications have expanded beyond the monogenic disease and chromosome aberration, as late-onset disease or HLA matching for stem cell donor.
Abortion, Induced ; Abortion, Spontaneous ; Aneuploidy ; Chromosome Aberrations ; Family Characteristics ; Female ; Humans ; Mass Screening ; Polymerase Chain Reaction ; Pregnancy ; Preimplantation Diagnosis* ; Prognosis ; Prostaglandins D ; Stem Cells ; Tissue Donors ; Translocation, Genetic