Effect of Angelica dahurica coumarins on the transport behavior of puerarin across blood-brain barrier in vitro and in vivo / 药学学报

A BBB co-culture cell model consisting of rat brain microvascular endothelial cells (BMEC) and astrocytes (AS) was established to study the effect of Angelica dahurica coumarins on the transport behavior of puerarin across blood-brain barrier (BBB) in vitro and in vivo. The barrier function of this model was evaluated by measuring the transendothelial resistance, phenol red permeability and BBB related protein expression. The permeability assay and western blot methods were performed to study the effects of Angelica dahurica coumarins on the BBB permeability and the expression of BBB related protein. The animal experiment protocols in this study were approved by the Animal Ethics Committee of Xi'an Jiaotong University (Animal Ethics No.: 2021-1329). The results showed that the established BMEC/AS co-culture model could be used to evaluate drug transport across BBB in vitro. After combined with Angelica dahurica coumarins, the transport capacity of puerarin was significantly increased in vitro and in vivo. Additionally, Angelica dahurica coumarins enhanced BBB permeability and inhibited the protein expression of P-glycoprotein (P-gp), zonula occludens-1 (ZO-1) and occludin. Angelica dahurica coumarins might increase BBB permeability by inhibiting the expression of P-gp and tight junction protein, thereby increasing the content of puerarin in brain tissue.

Ovarian Response and Pregnancy Rate according to Basal FSH concentration in Controlled Ovarian Hyperstimulation and Clomiphene Citrate stimulated cycle for ART / 대한산부인과학회잡지

OBJECTIVE: To evaluate whether elevation of basal FSH predict poor ovarian response and lowered pregnancy rate in women undergoing controlled ovarian hyperstimulation (COH) and Clomiphene Citrate stimulated cycle (CC cycle) for assisted reproductive technologies (ART). METERIALS AND METHODS: From January 1999 to December 1999, total 1067 COH cycles and 119 CC cycles from 1033 patients were included in this study. At each cycle, on cycle day 2 or 3, basal FSH was measured before GnRH agonist starting. FSH value (mIU/ml) was 2 nd IRP 78/549 standard. We divided COH and CC cycles into 4 groups according to elevated basal FSH concentration, respectively. i) Normal (Basal FSH OR =20 mIU/ml): Group D (n=35), Group IV (n=34). Retrospectively, we obtained mean total ampules of gonadotropin, mean serum E2 concenturation on hCG day, mean number of retrieved oocyte, mean number of embryo transferred, mean number of good embryo, cancellation rate, clinical pregnancy rate and live birth rate. RESULTS: Ovarian response by elevation of basal FSH decreased more significantly in COH cycles than CC cycles. In COH cycles, ovarian response of Group B, C and D decreased significantly (P<0.001). In CC cycles, ovarian response of Group IV decreased significantly (P<0.01). Including cycles only under 35 years old, COH cycles with mildly elevated basal FSH had poor ovarian response (P<0.01), but the clinical pregnancy rate (28.3%) and live birth rate (24.2%) did not decrease, compared with normal FSH Group (27.5%, 23.1% respectively). In cycles with markedly elevated basal FSH, clinical pregnancy rate (5.9%) and live birth rate (2.9%) of CC cycles were equal to that of COH cycles (5.7%, 2.9%, respectively). CONCLUSION: Mildly elevated basal FSH does not predict poor outcome in ART. Poor prognosis conferred by mildly elevated basal FSH may be overcome by maximizing stimulation protocol. Therefore other stimulation protocol for poor ovarian response may be effective in mildly elevated basal FSH cycles. In cycles with moderately to severe elevated basal FSH, lowered pregnancy rate was mainly due to quantitative and qualitative decrease in ovarian response. In cycles with markedly elevated basal FSH, CC stimulated cycle was more cost effective with good compliance.

Ovarian Response and Pregnancy Rate according to Basal FSH concentration in Controlled Ovarian Hyperstimulation and Clomiphene Citrate stimulated cycle for ART / 대한산부인과학회잡지

OBJECTIVE: To evaluate whether elevation of basal FSH predict poor ovarian response and lowered pregnancy rate in women undergoing controlled ovarian hyperstimulation (COH) and Clomiphene Citrate stimulated cycle (CC cycle) for assisted reproductive technologies (ART). METERIALS AND METHODS: From January 1999 to December 1999, total 1067 COH cycles and 119 CC cycles from 1033 patients were included in this study. At each cycle, on cycle day 2 or 3, basal FSH was measured before GnRH agonist starting. FSH value (mIU/ml) was 2 nd IRP 78/549 standard. We divided COH and CC cycles into 4 groups according to elevated basal FSH concentration, respectively. i) Normal (Basal FSH OR =20 mIU/ml): Group D (n=35), Group IV (n=34). Retrospectively, we obtained mean total ampules of gonadotropin, mean serum E2 concenturation on hCG day, mean number of retrieved oocyte, mean number of embryo transferred, mean number of good embryo, cancellation rate, clinical pregnancy rate and live birth rate. RESULTS: Ovarian response by elevation of basal FSH decreased more significantly in COH cycles than CC cycles. In COH cycles, ovarian response of Group B, C and D decreased significantly (P<0.001). In CC cycles, ovarian response of Group IV decreased significantly (P<0.01). Including cycles only under 35 years old, COH cycles with mildly elevated basal FSH had poor ovarian response (P<0.01), but the clinical pregnancy rate (28.3%) and live birth rate (24.2%) did not decrease, compared with normal FSH Group (27.5%, 23.1% respectively). In cycles with markedly elevated basal FSH, clinical pregnancy rate (5.9%) and live birth rate (2.9%) of CC cycles were equal to that of COH cycles (5.7%, 2.9%, respectively). CONCLUSION: Mildly elevated basal FSH does not predict poor outcome in ART. Poor prognosis conferred by mildly elevated basal FSH may be overcome by maximizing stimulation protocol. Therefore other stimulation protocol for poor ovarian response may be effective in mildly elevated basal FSH cycles. In cycles with moderately to severe elevated basal FSH, lowered pregnancy rate was mainly due to quantitative and qualitative decrease in ovarian response. In cycles with markedly elevated basal FSH, CC stimulated cycle was more cost effective with good compliance.

The efficacy of prophylactic intravenous albumin in clinical progress in patients with severe ovarian hyperstimulation syndrome who needed hospitalization / 대한산부인과학회잡지

OBJECTIVE: The purpose of this study is to evaluate the efficacy of prophylactic intravenous albumin in patients with severe ovarian hyperstimulation syndrome (OHSS) who needed hospitalization. METHODS: From January, 1995 to December, 2000, 86 women who underwent COH for IVF-ET were hospitalized for the management of severe OHSS in Samsung Cheil hospital. Among them, twenty one patients were treated with prophylactic intravenous albumin at the day of ovum retrieval, and 65 were not. We analysed clinical symptoms, signs, and hospital courses in the records of 86 patients retrospectively. We compared above variables of prophylactic albumin used group (n=21) with those of non-used group (n=65). RESULTS: Among the indices of ovarian response to ovarian stimulation, serum estradiol (E2) level at the day of hCG injection was significantly higher in prophylactic albumin used group (5805.2+/-1604.4 pg/ml) than non-used group (4125.2+/-1921.5 pg/ml, P<0.05). But the variables which indicate clinical progress (number and amount of paracentesis, amount of used albumin after hospitalization, duration of symptoms, hospital days) showed less severe tendency in prophylactic albumin used group but was not significantly different between two groups. CONCLUSION: Our results indicates that the use of prophylactic intravenous albumin in patients who have risk of severe OHSS had some advantages in favorable clinical progress.

Chromosomal Analysis of Aborted Conceptuses among Anembryonic Pregnancy and Embryonic Pregnancy: Do Chromosomal Abnormalities Relate to Anembryonic Pregnancy? / 대한산부인과학회잡지

OBJECTIVES: Approximately 15-20% of all clinically recognized pregnancies result in spontaneous abortion between 8 and 12 weeks. In some early abortus only the extra-embryonic components of the conceptus survive, either as fragments of placenta or as empty gestational sac, and these have been termed 'anembryonic pregnancies'. Despite their common occurrences, the etiology of anembryonic conceptuses is poorly understood. The purpose of this study was to evaluate whether chromosomal abnormalities relate to a role in anembryonic pregnancy. MATERIALS AND METHODS: The study, that consisted of 143 pregnancies from chromosomally normal couples that aborted in first trimester period between January 1, 1994 and December 27, 1997, had chromosomal analysis performed on the products of conception and had ultrasonographic examination prior to spontaneous abortion. The tissue was processed and analysed using standard G-banding cytogenetic techniques with long-term cultures. RESULTS: The patient characteristics of the anembryonic pregnancy were not significantly different from those of the embryonic pregnancy. Among products of conception from embryonic pregnancies, 45% (52/115) had normal chromosomal analyses and 55% (63/115) had abnormal chromosomal analyses. Products of conception from anembryonic pregnancies resulted in 54% (15/28) of abnormal chromosome analyses and normal chromosomal analyses resulted in 46% (13/28) (p>0.05, Chi-square test). The frequency of aneuploidy and polyploidy were not different between products of conception from anembryonic pregnancy and embryonic pregnancy. Also, the gender ratio of euploid spontaneous pregnancy losses showed similar in this study. CONCLUSION: There are no differences in frequency of abnormal karyotypes in products of conception from anembryonic pregnancies compared with miscarriage after the demonstration of fetal pole. If further studies are need for the evaluation of these etiologies, and then immuno-molecular studies in early placental tissues from anembryonic pregnancy might be helpful.

Outcome of Preimplantation Genetic Diagnosis for Chromosome Aneuploidy and Genetic Disease / 대한불임학회지

OBJECTIVES: Chromosome aneuploidy is associated with recurrent abortion and congenital anomaly and genetic diseases occur repeatedly in the specific families. Preimplantation genetic diagnosis (PGD) can prevent aneuploidy or genetic disease by selecting normal embryos before implantation and is an alternative to prenatal diagnosis. The aim of this study is to assess the outcome of PGD cycles by using FISH or PCR, and to determine the clinical usefulness and values in patients with risk of chromosomal aneuploidy or genetic disease. MATERIALS AND METHODS: From 1995 to Apr. 2001, a total of 108 PGD cycles in 65 patients with poor reproductive outcome were analyzed. The indications of PGD were translocation (n=49), inversion (n=2), aneuploidy screening (n=7), Duchenne muscular dystrophy (n=5) and spinal muscular atrophy (n=2). PGD was applied due to the history of recurrent abortion, previous birth of affected child or risk of aneuploidy related to sex chromosome aneuploidy or old age. Blastomere biopsy was performed in 6~10 cell stage embryo after IVF with ICSI. In the single blastomere, chromosome aneuploidy was diagnosed by using FISH and PCR was performed for the diagnosis of exon deletion in DMD or SMA. RESULTS: The FISH or PCR amplification was successful in 94.3% of biopsied blastomeres. The rate of transferable balanced embryos was 24.0% in the chromosome translocation and inversion, 57.1% for the DMD and SMA, and 28.8% for the aneuploidy screening. Overall hCG positive rate per transfer was 17.8% (18/101) and clinical pregnancy rate was 13.9% (14/101) (11 term pregnancy, 3 abortion, and 4 biochemical pregnancy). The clinical pregnancy rate of translocation and inversion was 12.9% (11/85) and abortion rate was 27.3% (3/11). In the DMD and SMA, the clinical pregnancy rate was 33.3% (3/9) and all delivered at term. The PGD results were confirmed by amniocentesis and were correct. When the embryos developed to compaction or morula, the pregnancy rate was higher (32%) than that of the cases without compaction (7.2%, p<0.01). CONCLUSIONS: PGD by using FISH or PCR is useful to get normal pregnancy by reducing spontaneous abortion associated with chromosome aneuploidy in the patients with structural chromosome aberration or risk of aneuploidy and can prevent genetic disease prior to implantation.

Outcome of Preimplantation Genetic Diagnosis for Chromosome Aneuploidy and Genetic Disease / 대한불임학회지

OBJECTIVES: Chromosome aneuploidy is associated with recurrent abortion and congenital anomaly and genetic diseases occur repeatedly in the specific families. Preimplantation genetic diagnosis (PGD) can prevent aneuploidy or genetic disease by selecting normal embryos before implantation and is an alternative to prenatal diagnosis. The aim of this study is to assess the outcome of PGD cycles by using FISH or PCR, and to determine the clinical usefulness and values in patients with risk of chromosomal aneuploidy or genetic disease. MATERIALS AND METHODS: From 1995 to Apr. 2001, a total of 108 PGD cycles in 65 patients with poor reproductive outcome were analyzed. The indications of PGD were translocation (n=49), inversion (n=2), aneuploidy screening (n=7), Duchenne muscular dystrophy (n=5) and spinal muscular atrophy (n=2). PGD was applied due to the history of recurrent abortion, previous birth of affected child or risk of aneuploidy related to sex chromosome aneuploidy or old age. Blastomere biopsy was performed in 6~10 cell stage embryo after IVF with ICSI. In the single blastomere, chromosome aneuploidy was diagnosed by using FISH and PCR was performed for the diagnosis of exon deletion in DMD or SMA. RESULTS: The FISH or PCR amplification was successful in 94.3% of biopsied blastomeres. The rate of transferable balanced embryos was 24.0% in the chromosome translocation and inversion, 57.1% for the DMD and SMA, and 28.8% for the aneuploidy screening. Overall hCG positive rate per transfer was 17.8% (18/101) and clinical pregnancy rate was 13.9% (14/101) (11 term pregnancy, 3 abortion, and 4 biochemical pregnancy). The clinical pregnancy rate of translocation and inversion was 12.9% (11/85) and abortion rate was 27.3% (3/11). In the DMD and SMA, the clinical pregnancy rate was 33.3% (3/9) and all delivered at term. The PGD results were confirmed by amniocentesis and were correct. When the embryos developed to compaction or morula, the pregnancy rate was higher (32%) than that of the cases without compaction (7.2%, p<0.01). CONCLUSIONS: PGD by using FISH or PCR is useful to get normal pregnancy by reducing spontaneous abortion associated with chromosome aneuploidy in the patients with structural chromosome aberration or risk of aneuploidy and can prevent genetic disease prior to implantation.

Obstetric outcome in women with a history of recurrent miscarriage / 대한산부인과학회잡지

OBJECTIVE: To obtain etiology and obstetric outcome of women who had a history of recurrent miscarriage. METHODS: From 1 June 1998 to 30 June 2000, 82 patients who attended infertility & current miscarriage clinic at Samsungcheil hospital and progressed beyond 24 weeks gestation following pregnancy were included in this study. The control population was 154 pregnancies considering age and parity over the same period. Retrospectively, we analyse the etiology of recurrent miscarriage and compare obstetric outcomes of two groups. RESULTS: The etiology of recurrent miscarriage was immunologic factor (36.6%), unexplained (30.4%), anatomic cause (13.4%), endocrinologic abnormality (13.4%) and chromosomal abnormality (5.6%). The rate of preterm delivery (11%) and incidence of pregnancy induced hypertention (8.5%) were significantly higher than those of control group (3.2% and 2.5% respectively). There was no significant difference in the rate of small for gestational age, oligohydroamnios, cesarean section, perinatal loss and the incidence of gestational diabetes mellitus. CONCLUSION: The pregnancy with a history of recurrent miscarriage is associated with increased risk of pregnancy induced hypertension and preterm delivery and represent a population at high risk of obstetric problems. Therefore, close surveillance during antenatal period is required.

Influence on the Pregnancy Rate of Motility of Fresh and Frozen Testicular Spermatozoa in Obstructive Azoospermic Patients / 대한불임학회지

OBJECTIVE: ICSI with testicular sperm could achieve optimal fertilization and pregnancy. This study was performed to observe the influence on fertilization and pregnancy of motility of fresh testicular sperm and sperm extracted from frozen-thawed seminiferous tubules in obstructive azoospermia. MATERIALS ANDMETHODS: We analysed clinical outcome of ICSI using fresh testicular sperm and sperm extracted from thawed seminiferous tubules. The presence of motility were compared to determine the factor for optimal fertilization and pregnancy rates. RESULTS: In 316 cases of TESE-ICSI in obstructive azoospermia, ICSI with fresh testicular sperm (fresh sperm group) were 163 cases and ICSI with sperm testicular sperm extracted from frozen-thawed seminiferous tubule (thawed sperm group) were 153 cases. The fertilization rates were 71.3% and pregnancy rates were 32.5% in fresh sperm group, in thawed sperm group, 65.1% and 33.3% respectively. The fertilization and pregnancy rates of motile and non-motile testicular sperm were 72.9% and 33.6%, 50.0% and 18.2%, respectively (p<0.05). The fertilization and pregnancy rates of motile and non-motile sperm extracted from the thawed seminiferous tubule were 67.8% and 34.7%, 55.1% and 28.1%, respectively (p<0.05). The comparative of the results of ICSI using motile fresh testicular sperm and motile sperm extracted from thawed seminiferous tubule, fertilization and pregnancy rates were not significantly different (72.9% and 33.6%, 67.8% and 34.7%, respectively). CONCLUSION: These results suggest that successful pregnancy in TESE-ICSI treatment is influenced by the motility of fresh testicular sperm and sperm extracted from thawed seminiferous tubule in obstructive azoospermic patients.

Expression of leukemia inhibitory factor in endometrium of infertile women during the window of implantation / 대한산부인과학회잡지

OBJECTIVE: To determine the expression of endometrial leukemia inhibitory factor (LIF) in infertile women and to compare it with that of fertile women during the window of implantation METHODS: Endometrial biopsies were performed during the window of implantation from 10 normal fertile and 15 infertile women (5 unexplained infertility, 5 endometriosis and 5 tubal disease with hydrosalpinx). We performed immunohistochemistry using paraffin-embedded endometrial tissues. RESULTS: In fertile and infertile women, the staining intensities of LIF in the glandular epithelium were significantly higher (p<0.01) than those of stromal cells. There was no significantly difference in the expression of LIF between the glandular epithelium of infertile women with endometriosis or hydrosalpinx and those of fertile women. However, the staining intensities of LIF in the glandular epithelium of unexplained infertility patients were significantly lower (p<0.01) than those in fertile women. The staining intensities of LIF in stromal cell of women with unexplained infertility, endometriosis and hydrosalpinx were similar to those in fertile women. CONCLUSIONS: This study demonstrated that the glandular epithelial cells are more important sites of LIF secretion than stromal cells. The expression of endometrial LIF may be decreased in women with unexplained infertility during the window of implantation. Therefore, this result suggest that abnormalities of LIF expression in infertile women may underlie endometrial dysfunction in the adhesive phase of implantation.

The clinical efficacy of medical treatment and / or laparoscopic surgery for the improvement of pregnancy rate after IVF - ET in the infertile patients with endometriosis / 대한산부인과학회잡지

OBJECTIVE: The aim of study was to evaluate the efficacy of medical and/or laparoscopic surgical treatment for improvement of reproductive outcome of in vitro fertilization and embryo transfer (IVF-ET) in the patients with endometriosis. METHODS: 296 IVF-ET cycles except 18 cycles of cancelled embryo-transfer (unfertilization; 16 cycles, high risk of ovarian hyperstimulation syndrome; 2 cycles) in 191 infertile women with endometriosis from January 1, 1995 to December 31, 1998 were included in this study. All women's ages were < or = 35 and the factor for infetility was endometriosis only and day 3 follicle stumulating hormone (FSH) level of all the cycles was < or = 10 mIU/ml in this study. All the baseline study about infertility was done before initiating treatment of infertility. The stage of endometriosis was recorded during diagnostic laparoscopy by WHO critera. Medical treatment (GnRH agonist or danazole) or laparoscopic surgery was done independently or combinedly by the physician's decision according to the disease entity and symptoms. Short or long protocols were used for controlled ovarian hyperstimulation and 3 day embryos were transferred (maximally 4 embryos) by standard procedures in our institute. We classified the cases as 4 groups : group 1 (without pre-treatment, 80 cycles), goup 2 (laparoscopic surgery only, 37 cycles), group 3 (medication only, 140 cycles), group IV (combined pre-treatment, 39 cycles). RESULTS: The mean age (years old, mean +/- SEM) was 31.5 +/- 0.3 (group 1), 31.6 +/- 0.5 (group 2), 31.5 +/- 0.2 (group 3), 31.7 +/- 0.4 (group 4) respectively. The duration of infertility (months) was 57.7 +/- 3.3, 64.5 +/- 4.8, 59.1 +/- 1.9, 52.0 +/- 3.7 respectively among groups. The viable pregnancy rate (over 28 gestational weeks, VPR) was 12.5 % (10/80) in goup 1, 13.5 % (5/37) in goup 2, 14.3 % (20/140) in group 3, 30.8 % (12/39) in group 4 and there was statistically significant difference between group 1 and group 4 (P=0.03, Chi square test). The total used gonadotropins (ampules) for controlled ovarian hyperstimulation were 28.2 +/- 0.6.

Three cases of successful pregnancy outcome after intensive care in the patients with complete uterine septum, cervical duplication and longitudinal vaginal septum / 대한산부인과학회잡지

Three cases of successful pregnancy outcome after intensive care in the patients with rare mullerian anomaly of complete uterine septum, hypoplastic cervical duplication and longitudinal vaginal septum in infertility clinic are reported. These presented three cases are the reports of supporting the embryologic hypothesis of bidirectional mullerian development which fusion and resorption begins at the isthmus and proceeds simultaneously in both the cranial and caudal directions rather than the classical views of unidirectional(caudal to cranial) mullerian development. Thus, we present here three cases of rare mullerian anomaly and their pregnancy outcomes in an attempt to alert gynecologist to the possible occurrence of such a malformation. In 2 cases, successful pregnancy outcomes were achieved after abdominal or hysteroscopic uterine septotomy before pregnancy and in the last case, successful pregnancy outcome intensive obstetric care was achieved even under uncorrected uterine anomaly and complete removal of uterine septum resulted in uneventful postoperative courses at the same time of cesarean section.

Analysis of Factors Affecting Survival and Pregnancy Rate in Frozen-thawed Embryo Transfers / 대한불임학회지

OBJECTIVE: The purpose of this study was to determine the important factors affecting survival and pregnancy rate in frozen-thawed embryo transfer cycles. METHODS: we performed reprospective analysis in 738 cycles of frozen-thawed embryo transfers, in relation to the insemination methods, the freezing stage of embryo, patient's age, infertility factors and the origin of injected sperm in ICSI cycles. After conventional IVF or ICSI, the supernumerary PN stage zygotes or multicellular embryos were cryopreserved by slow freezing protocol with 1,2-propandiol (PROH) as a cryoprotectant. RESULTS: The survival rates of thawed embryos were 69.3% (1585/2287) in conventional IVF group and 71.7&% (1645/2295) in ICSI group. After frozen-thawed embryo transfers, 27.0% (92/341) and 32.0% (109/341) of pregnancy rates were achieved in conventional IVF and ICSI group, respectively. There were no significant difference in the survival and pregnancy rates according to the insemination methods, the freezing stage and patient's age. However, the pregnancy rate (36.2%) of male factor infertility (22.9%). In ICSI group, the origin of injected sperm did not affect the outcome of frozen-thawed embryo transfer cycles. CONCLUSION: The present study demonstrates that acceptable clinical outcomes can be achieved after the transfer of frozen-thawed embryos regardless of the stage of embryos for freezing, the patient's age and the origin of injected sperm.

Development of Inclusion Bodies in Type II Pneumocyte of Human Fetus / 체질인류학회지

The formation and development of cytoplasmic inclusion bodies of type II pneumocyte were investigated using 7 cases of human fetal lungs from 9 to 20 weeks of gestation by transmission electron micropscopy. The results obtained were as follows: 1. The multilamellar bodies, the characteristic inclusion body of type II pneumocyte, have developed in developing epithelium of lung at 9 week of gestation. Another inclusion bodies specific to type II pneumocyte also have developed at that time. 2. The inclusion bodies were formed in association with cytoplasmic reticulum, outer membrane of nuclear envelope, and mitochondria. 3. The inclusion bodies were distributed in cluster at the apical cytoplasm, and classified schematically with the contents as multilamellar, cytoplasmic, granular/f locculent, multivesicular, dense, and multilamellar. But the intermediate and composite forms of inclusion bodies appeared at the time toward 20 week of gestation. In summary, it is suggested that the differentiation of type II pneumocyte starts before 9 week of gestation and 4 main types of inclusion bodies considered as the precursor of multilamellar body were found. Although the inclusion bodies were formed at endoplasmic reticulum or etc, it is likely that they form multilamellar body through the complex process such as fusion of inclusion bodies.

Adverse Effect of Human Hydrosalpingeal Fluid on the Development of Mouse Embryo (II) / 대한불임학회지

In our previous study, we observed that hydrosalpingeal fluid (HSF) adversely effect mouswe embryo development and hatching. The aim of this study was to evaluate the effect of HSF as assessed by the blastocyst development rate (BDR) and by cell counting in vitro HSF was collected from nine patients undergoing salpingoneostomy to correct hydrosalpinx. Two-cell embryos were obtained from superovulated ICR mice. T6 medium and T6+/-0.4% bovine serum albumin were used as control media. T6 medium containing 10% or 50% HSF and 100% HSF from each patient were used as test media. Nine to 15 embryos were cultured in microdrops prepared from each of these media. To assess the total cell number within each blastocyst, the blastocysts were fixed and stained with Hoechst 33342 to facilitate cell counting. The mean BDR in two control media were 88.89% and 85.40%. The mean BDR in media containing 10%, 50%, 100% HSF were 85.87%, 89.58% and 75.57%, respectively (*: p<0.05). The overall mean cell count (+/-SEM) in control media were 87.6+/-9.65 and 90.12+/-11.38. The BDR was affected adversely only by 100% HSF and not in media containing 10% or 50% HSF. Mean cell counts were decreased significantly only in blastocysts cultured 100% HSF (63.8+/-13.66; p<0.01) but not in blastocysts cultured in 10% or 50% HSF (91.3+/-12.44 and 82.9+/-18.27, respectively). Thus, it is concluded that HSF has no embyotoxic effect but has a mildly negatively effect on embryonic growth and development.

Adhesive potential of the shed endometrial cells in menstrual fluid using amniotic membrane: establishment of in-vitro model for pathogenesis of endometriosis / 대한산부인과학회잡지

OBJECTIVES: The purpose of this study was to determine whether or not the shed endometrial tissues in menstrual fluid (MF) have adhesive potentials, using human amniotic membrane (AM). METHODS: The MF from 20 patients with regular menstruation was collected with Wallace catheter by aspiration from the uterine cavity on the second or third day of the menstrual period. The AM was obtained from the placenta of term delivery without any complication. The MF was washed and diluted fivefold with Hams F-10 medium supplemented with 10% fetal bovine serum. The cell suspension was placed on either epithelial layer (EP) or extracellular matrix layer (ECM) of the AM. After 5 days of culture, the adhesion sites were observed under a stereomicroscope. For histological observation, each cultured AM was prepared for the serial paraffin section. RESULTS: The adhesion sites of endometrial tissues in MF were found both ECM (20/20) and EP (11/20) of the AM. The size of adhesion sites in each AM were highly variable from microscopic to macroscopic size. CONCLUSION: We found that the shed endometrial tissues in MF have adhesive potential to epithelial layer in addition to extracellular matrix layer of amniotic membrane. This adhesive potential may be related to pathogenesis of endometriosis. We suggest that this culture system can be useful as an in-vitro model for endometriosis.

Prediction and Clinical Evaluation of Hyperstimulation Syndrome / 대한산부인과학회잡지

OBJECTIVE: Ovarian hyperstimulation syndrome (OHSS) is one of the well known complication of conttolled ovarian hyperstimulation. Though there have been numerous protocols for the prevention of OHSS, it has not been completely preventable until now. This study was performed to identify clinical predictors for early and late OHSS. METHODS: A retrospective analysis of all IVF cycles in 1993 up to June 1996 was performed. OHSS was diagnosed using the criteria of Rabau modified by Schenker. All cases of OHSS reported in this study presented with marked ovarian enlargement, ascites, oliguria, hemoconcentration and electrolyte disturbance. Ovarian stimulation was carried out using a combination of gonadotrophin releasing hormone-agonist, follicle-stimulation hormone and human menopausal gonadotrophin. 27 patients has moderate or severe OHSS presenting 3-7 days post-human chorionic gonadotrophin (hCG), and 21 patients had severe OHSS presenting 12-17 days post-hCG. RESULTS: No patient with early OHSS went onto develop late OHSS, and no patient with late OHSS had demonstrated early OHSS. Logistic regression showed that early OHSS was predicted by the number of oocytes retrieved and the estradiol concentration on the day hCG injection (P<0.05). Late OHSS was predicted by the transferred embryos, B-hCG on 14 day after hCG injection (P<0.05). CONCLUSION: Early OHSS was an acute effect of the hCG administered prior to egg retrieval in women with high estradiol and large number of retrieved oocytes. Our analysis of the risk factors for early OHSS indicates that cryopreservation of all embryos will not alter the risk of early OHSS even though it should prevent late OHSS. Late OHSS was induced by the rising serum concentration of hCG produced by the early pregnancy, the number of transferred embryos must be adjusted carefully, since it was associated with multiple gestation.

Reproductive Outcome of Women with Recurrent Abortions or Infertility Following Treatment by Operative Hysteroscopy for an Intrauterine Septum / 대한산부인과학회잡지

Uterine anomalies have been reported in 4% of women with infertility and in up to 15% of those with recurrent abortion. One of the major intrauterine disorder associated with infertility and recurrent abortions is intrauterine septum, The reproductive outcome of 41 patients of intrauterine septum (7 complete, 34 incomplete) with repeated abortions or infertility was assessed after the uterine septotomy. 5 of 7 patients with comlete uterine septum undergone uterine septotomy (3; hysteroscopic metroplasty, 2; abdominal metroplasty) had total 6 pregnancies and all of them had live biths. 28 patients with incomplete uterine septum got the hysteroscopic intrauterine septotomy and the viable pregnancy rate was 62% (3 ongoing pregnancies, 13 live biths of total 26 pregnancies). 6 patients with incomplete uterine septum had not the operation and 5 patients had 5 live births after total 6 pregnancies with 1 spontaneus abortion. Even though, the number of cases were small, the live birth rate in the group of septotomy of the patients of complete uterine septum (100%, 6/6) was higher than that in the group of not-done (50%, 1/2). The live birth rate in the group of not-done of the patients with incomplete uterine septum (83%, 5/6) was higher than that in the group of hysteroscopic uterine septotomy (62%, 16/26), but 5 of 6 had short uterine septal length (<1 cm), 1 had 1.5 cm septal length in the group of not-done. All the patients with successful pregnancy outcome had no other co-factors at the diagnostic laparoscopy, but the 5 primary infertility patients with no live birth even after treatment (all were with incomplete septum; 3 undergone hysteroscopic septotomy, 2 not-done with one abortion) had other co-factors such as endometriosis, peritoneal or tubal facor. In conclusion, hysteroscopic uterine septotomy would be useful for the patients with habitutal abortion or infertility and more advanced managemnet protocols should be applied to the patients having other co-factors if there was no pregnancy even after the uterine septotomy.

Efficacy of Clomiphene Citrate Stimulated Cycle in Poor Responders in inviro Fertilization / 대한불임학회지

The purpose of this study is compare IVF cycle outcome in poor responders between clomiphene citrate (CC) stimulated and controlled ovarian hyperstimulation (COH) protocol. A total of 94 patients responding poorly in previous IVF cycles (estradiol16 mm. In COH group, ovarian stimulation was done using short protocol (GnRH-a/FSH/HMG/hCG). No difference in age or number of transferred embryos was found between CC group and COH group. COH group had significantly (p<0.05) higher mean peak level of E2 (810+/-112 vs 412+/-55 pg/ml) and greater number of retrieved oocytes (3.0+/-0.2 vs 2.0+/-0.2) than CC group. CC group had significantly embryos (1.8+/-0.2) compared with (2.1+/-0.2) in COH group. However, CC group had higher pregnancy rate than COH group per retrieval [26.9% (7/26) vs 6.2% (6/97)], or per transfer [31.8% (7/22) vs 7% (6/86)]. Although cycle cancellation rate in CC group (48.8%) was higher than that of COH group (21.3%), the pregnancy rate per cycle in CC group was still higher (16.3%) than COH group (4.9%). In addition, implantation rate in CC group was 17.5% (7/40), which was significantly p<0.01) higher than 3.9% (7/180) in COH group. These data suggest that oocyte and embryo quality are lower in COH cycles of poor responders than CC cycles. We suggest that clomiphene citrate stimulated IVF cycle may be more efficient than COH IVF cycle in poor responders in terms of lower costs and higher pregnancy performance.

Comparative Study of Perinatal Outcome and the Incidene of Congenital Anomalies of Babies Born after Intracytoplasmic Sperm Injection (ICSI) and Conventional In-vitro Fertilization (IVF) / 대한불임학회지

The safety of ICSl as a novel procedure of assisted fertilization may be assessed by the health of the baby born. In order to evaluate the safety of ICSI, perinatal outcome and congenital anomaly of the babies born after ICSI were compared with those of babies born after IVF (control group). We analysed the clinical data from the obstetric and pediatric records, including the information obtained through telephone. The results are as follows; Mean gestaional age (+/-SEM) and birth weight in singleton pregnancy were 38.8+/-1.9 weeks and 3209.7+/-501.9gm in IVF group, 39.0+/-2.2 weeks and 3289.9+/-479.5gm in ICSI group, respectively. Mean gestational age and birth weight in twins were 36.8+/-2.1 weeks and 2512.8+/-468.0gm in IVF group, 36.5+/-2.8 weeks and 2492.7+/-537.1gm in ICSI group. In IVF group, perinatal mortality rates were 8.5 in singletons and 56.6 in twinst for the ICSI singletons and ICSI twins, the perinatal mortality rates were 11.6 and 49.0, respectively. The incidence of congenital malformations was 3.6% (8/224) in IVF group and 2.1% (4/188) in ICSI group, there was no statistical difference (p>0.05, Fisher's exact test). The incidence of major congenital anomalies was 0.9% (2/224; pulmonary artery hypoplasia, renal cystic dysplasia) in IVF group and 1.1% (2/188; holoprosencephaly, Cri du chat syndrome) in ICSI groups (p>0.05, Fisher's exact test). Similarly, there was no significant difference in incidence of minor congenital anormalies 2.7% (6/224) in IVF group and 1.1% (2/188) in ICSI group respectively (p>0.05, Fisher's exact test). In conclusion, there was no difference in the perinatal outcome and the incidence of congenital anomalies between the babies born after ICSI and those after conventional IVF.