Current status of assisted reproductive technology in Korea, 2009

Great advances have been made in the field of assisted reproductive technology (ART) since the first in vitro fertilization (IVF) baby was born in Korea in the year of 1985. However, it deserve to say that the invaluable data from fertility centers may serve as a useful source to find out which factors affect successful IVF outcome and to offer applicable information to infertile patients and fertility clinics. This article intended to report the status of ART in 2009 Korean Society of Obstetrics and Gynecology surveyed. The current survey was performed to assess the status and success rate of ART performed in Korea, between January 1 and December 31, 2009. Reporting forms had been sent out to IVF centers via e-mail, and collected by e-mail as well in 2012. With International Committee Monitoring Assisted Reproductive Technologies recommendation, intracytoplasmic sperm injection (ICSI) and non-ICSI cases have been categorized and also IVF-ET cases involving frozen embryo replacement have been surveyed separately. Seventy-four centers have reported the treatment cycles initiated in the year of 2009, and had performed a total of 27,947 cycles of ART treatments. Among a total of 27,947 treatment cycles, IVF and ICSI cases added up to 22,049 (78.9%), with 45.3% IVF without ICSI and 54.7% IVF with ICSI, respectively. Among the IVF and ICSI patients, patients confirmed to have achieved clinical pregnancy was 28.8% per cycle with oocyte retrieval, and 30.9% per cycle with embryo transfer. The most common number of embryos transferred in 2009 is three embryos (40.4%), followed by 2 embryos (28.4%) and a single embryo transferred (13.6%). Among IVF and ICSI cycles that resulted in multiple live births, twin pregnancy rate was 45.3% and triple pregnancy rate was 1.1%. A total of 191 cases of oocyte donation had been performed to result in 25.0% of live birth rate. Meanwhile, a total of 5,619 cases of frozen embryo replacement had been performed with 33.7% of clinical pregnancy rate per cycle with embryo transfer. When comparing with international registry data, clinical pregnancy rate per transfer from fresh IVF cycles including ICSI (34.1%,) was comparable to clinical pregnancy rate per transfer in European Society for Human Reproduction and Embryology report was 32.5% though lower than 45.0% for USA data. There was no remarkable difference in status of assisted reproductive technology in Korea between the current report and the data reported in 2008. The age of women trying to get pregnant was reconfirmed to be the most important factor that may have impact on success of ART treatment.

Identification of High Risk Group of HPV associated with Cervical Cancer in Korean Women / 대한산부인과학회잡지

OBJECTIVE: To identify genital HPV types and high risk group of HPV associated with cervical cancer in Korean women. METHODS: Both Pap test and HPV-DNA test using PCR assay were performed as screening test for cervical cancer in this clinic. When patients were positive in HPV-DNA test, HPV genotyping using sequencing method and cervical biopsy were performed. RESULTS: Frequent age group of HPV infection was 40 yrs (34.3%) and prevalence of HPV infection was 9.8%. Twenty-three types of HPV were detected. HPV 16 and 58 were detected in invasive cancer. HPV 16, 31, 33, 45, and 58 were detected in HSIL. HPV 6, 11, 18, 53, 59, and 66 were detected in LSIL. HPV 16 was most commonly detected in HSIL and invasive cancer. CONCLUSION: HPV 16, 31, 33, 45, and 58 are included in high risk group of HPV in Korean women. It may be very effective in early detection of cervical cancer to classify HPV types included in high risk group of cervical cancer in Korean women and to perform cervical biopsy in the patients who have high risk types of HPV infection.

Identification of High Risk Group of HPV associated with Cervical Cancer in Korean Women / 대한산부인과학회잡지

OBJECTIVE: To identify genital HPV types and high risk group of HPV associated with cervical cancer in Korean women. METHODS: Both Pap test and HPV-DNA test using PCR assay were performed as screening test for cervical cancer in this clinic. When patients were positive in HPV-DNA test, HPV genotyping using sequencing method and cervical biopsy were performed. RESULTS: Frequent age group of HPV infection was 40 yrs (34.3%) and prevalence of HPV infection was 9.8%. Twenty-three types of HPV were detected. HPV 16 and 58 were detected in invasive cancer. HPV 16, 31, 33, 45, and 58 were detected in HSIL. HPV 6, 11, 18, 53, 59, and 66 were detected in LSIL. HPV 16 was most commonly detected in HSIL and invasive cancer. CONCLUSION: HPV 16, 31, 33, 45, and 58 are included in high risk group of HPV in Korean women. It may be very effective in early detection of cervical cancer to classify HPV types included in high risk group of cervical cancer in Korean women and to perform cervical biopsy in the patients who have high risk types of HPV infection.

Comparison of IVF-ET Outcome after Various Therapeutic Approaches for Ovarian Endometriomas / 대한불임학회지

OBJECTIVE: To compare COH characteristics and IVF outcomes among IVF-ET patients who were treated with various therapeutic modalities for ovarian endometriomas and to propose effective pre-cyclic therapeutic modalities to improve IVF-ET outcomes in the patients with ovarian endometriomas. METHODS: All cases that had undergone IVF-ET after laparoscopy between January 1997 to August 2003 were reviewed. Forty-eight patients with tubal factor were assigned to Group I. Twenty seven, 22 and 38 patients diagnosed as severe pelvic adhesion with ovarian endometriomas by laparoscopy received only medical therapy (Group II), cyst aspiration (Group III), and sclerotherapy (Group IV), respectively. Laparoscopic cystectomy was performed in 20 patients (Group V). Resistance index was measured on day administering hCG. RESULTS: As compared with Group I, in Group II resistance index increased (p<0.05) but number of oocytes, good-quality oocyte ratio (mature and intermediate oocytes/total retrieval oocytes), fertilization rate, and embryo development rate decreased (p<0.05). In Group III fertilization rate and embryo development rate decreased (p<0.05). There was no difference between Group IV and Group I in all parameters except basal FSH which increased (p<0.05). In Group V basal FSH, and resistance increased (p<0.05) and number of oocytes and good-quality oocytes ratio decreased (p<0.05). CONCLUSION: Sclerotherapy is an effective therapeutic option which can be done prior to IVF-ET cycles in the patients with ovarian endometriomas. Further studies on a large scale are necessary to confirm these data.

Dietary Behavior of Infertile Women In Korea / 대한불임학회지

OBJECTIVE: The purpose of this study was to identify dietary factors related to infertility in Korean women through a case-control study. METHODS: The case group was composed of 236 women who had been diagnosed as infertility in hospital. The control group of 181 healthy women with children were recruited from local immunization centers. Socio-economic status, medical history, dietary intakes using food frequency questionnaire and stress were surveyed by interview. Anthropometric measurements were made and the causes of infertility were identified through medical records. Fasting blood samples were taken from subgroup of the subjects. RESULTS: The mean age of infertile and control groups was 31.1 and 32.4 years, respectively and the difference was statistically significant. The mean Body Mass Index of infertile women was not significantly different from control women, however, Waist/ Hip Ratio and Triceps Skinfolds Thickness were significantly lower in infertile women than in control women. The dietary intake status was generally satisfactory in both groups. The intakes of energy, protein, fat, carbohydrate, retinol, vitamin B2 and niacin were lower in infertile women than in control women. The infertile women also showed lower intakes of animal foods.No differences were found between two groups in serum concentrations of albumin, hemoglobin, Fe, TIBC, total cholesterol, HDL-cholesterol, LDL-cholesterol, triglyceride, C3, IgA, IL-2, however, infertile women showed higher levels of Zn and IgG. The stress score was higher in infertile women. CONCLUSIONS: From the results of this study, dietary factors and nutritional status do not seem to be directly related to infertility. However, the intertile women have lower nutrient intake and lower body fat content than control women. Further researches are needed according to the causes of infertility for long term to establish the relationship between dietary factors and infertility.

Effectiveness of Estradiol and Sildenafil supplement therapy in IVF-ET patients with thin endometrium / 대한산부인과학회잡지

OBJECTIVE: The purpose of this study was to evaluate the effect of estradiol (E2) only/and sildenafil supplement on controlled ovarian hyperstimulation and pregnancy outcome in patients undergoing IVF-ET cycles. METHODS: Total 104 cycles of IVF-ET patients were included in this study, which had been undergone controlled ovarian hyperstimulation (COH) with long protocol in Eulji infertility center and Ilsan Grace hospital between January 1999 and December 2000. Group 1 (COH only) consisted of 34 cycles, group 2 (COH+estradiol supplement) consisted of 35 cycles, and group 3 (COH+estradiol/sildenafil supplement) consisted of 35 cycles. RESULTS: With E2 and/only Sildenafil supplement, improvement of the endometrial thickness (P<0.05) and clinical pregnancy rate (P<0.05) were obtained. There was no significant increase in pregnancy rate with sildenafil supplement compared to E2 supplement only (20.0% versus 25.7%). However, rather short duration of controlled ovarian hyperstimulation (13.3+/-1.7 days versus 11.7+/-1.7 days) was found in the group that received E2 and sildenafil supplementation. (P<0.1) CONCLUSION: In patients with thin endometrium, the sildenafil supplement might lead to increase endometrial receptivity, and in consequence improvement of pregnancy rate. Sildenafil may augment the vasodilatory effect of nitric oxide by inhibition of cGMP hydrolysis, by improving ovarian perfusion, stimulates follicular development thus might shorten the duration of controlled ovarian hyperstimulation in IVT-ET cycles as we observed in our study.

Efficacy of estradiol supplement therapy for patients with thin endometrium in intrauterine insemination cycles / 대한산부인과학회잡지

OBJECTIVE: To evaluate the endometrial response and to compare the pregnancy outcome of estradiol supplement in patients with abnormally thin endometrium who are undergoing IUI. METHODS : From November 1st, 1998 to February 28th, 2001, 914 IUI cycles were studied and which were divided into several groups. In preparatory cycle, all of the patients were prepared with conjugated estrogen. The patients were divided into several groups according to the endometrial thickness (ET). Control I (n=734) was normal control group (ET>or=7 mm). Control II (n=67) was control group with abnormally thin endometrium (ETintrauterine insemination7 mm) without estradiol supplement. Group I (n=65) and group II (n=48) had thin endometrial thickness. However, in preparatory cycle, the endometrial thickness was more than 7 mm in group I and was less than 7 mm in group II. Uterine preparation consisted of 6-8 mg of estradiol valerate. The number of natural cycle was 234 and the hyperstimulation protocol used were clomiphene (n=250), clomiphene/ hMG (n=214), hMG (n=216). RESULTS: The average pregnancy rate in group I was 15.4%. There was no significant difference between control I (21.1%) and group I. The pregnancy rate in control II and group II was significantly decreased (3.0 vs. 6.3%) compared with control I and group I. In control I and group I, average endometrial thickness and pregnancy rate were decreased when clomiphene was used compared with hMG alone. (endometrial thickness control I 8.4 +/- 0.6 vs. 10.0 +/- 0.7 mm, group I 6.9 +/- 0.8 vs. 7.9 +/- 0.7 mm, pregnancy rate control I 14.6 vs. 29.8%, group I 9.1 vs. 31.3%). CONCLUSION: The adequate endometrial thickness is an important prognostic factor for implantation and is achieved with administration of estradiol supplement in patients with abnormally thin endometrium who responded to exogenous estradiol with endometrial thickness up to 7 mm in evaluation cycle.

Effect of Partial Laser Assisted Hatching on Mouse Embryos / 대한불임학회지

OBJECTIVE: The present study was performed to investigate the efficiency of partial laser assisted hatching (p-LAH; lased 1/2 ZP width from ZP edge) on hatching of mouse blastocysts. METHODS: We used non-contact 1.48 micrometer diode laser (MTM, Switzland) to create a precise hole on zona pellucida. 2-cell embryos were collected from the mouse (ICR) oviduct at 48 hours after hCG administration. Collected 2-cell embryos were cultured in the P-1 medium supplemented with 0.4% BSA. For experiments, embryos at 8-cell stage were used after 20~22 hours in culture. After conventional (c-LAH) or partial laser assisted hatching, the embryos were further cultured in P-1 medium supplemented with 0.4% BSA for 3 days. To compare efficiency of complete and partial laser assisted hatching, hatching rate, hatching time and blastocyst diameter and zona pellucida thickness at hatching time were investigated. Embryos were examined every 12 hours. Blastocyst diameter and zona pellucida thickness at hatching time were measured with an ocular micrometer. RESULTS: Hatching rates of p-LAH group (84.2%) was significantly higher than that of control group (39.3%), but there was no difference between the p-LAH (84.2%) and c-LAH (91.2%). p-LAH group was hatched 12 hours earlier than control group, but hatched 12 hours later than c-LAH group. The diameter of blastocyst at hatching time of p-LAH group (113.1+/-6.4 micrometer) was smaller than that of control group (122.2+/-5.0 micrometer), but larger than that of c-LAH group (102.2+/-2.7 micrometer). Zona pellucida thickness at hatching time of p-LAH group (6.4+/-0.9 micrometer) was thicker than that of control group (4.5+/-1.5 micrometer), but thinner than that of c-LAH group (10.0+/-0.8 micrometer). CONCLUSION: These results suggest that p-LAH may maintains the cell arrangement of early embryos to ensure successful development and prevent precocious hatching of blastocyst when compare to c-LAH and conventional (acidic tyrode) AH. Thus, p-LAH may provide a valuable and effective AH technique for human ART program.

The Effect of GM-CSF Supplementation in Culture Medium in the Human IVF Programs / 대한불임학회지

OBJECTIVE: Granulocyte-macrophage colony stimulating factors known to be secreted in murine and human reproductive tract. The development of human, bovine and murine embryos could be promoted by addition of GM-CSF in culture medium. However, the pregnancy and implantation rate of embryos cultured in GM-CSF have not been evaluated. The aim of this study was to assess the effect of GM-CSF in embryo development, pregnancy and implantation rate. METHODS: A total of 191 IVF cycles were divided into control and GM-CSF supplement group (control =96, GM-CSF=95). The embryos were cultured for three day with or without 2 ng/ml of recombinant human GM-CSF. The quality of embryo, developmental velocity, pregnancy and implantation rates were compared. RESULTS: There was no difference in age, number of gonadotropin ampules used, number of oocytes and fertilization. The number of ICSI cycle was higher in GM-CSF group. In GM-CSF group, G-1 grade embryos were the highest in proportion (56.4%), while G-2 grade embryos were highest (44.3%) in control group. The developmental velocity of embryos were not different between GM-CSF and control group. The pregnancy and implantation rates were significantly higher in GM-CSF group than control (47.4% vs. 33.3%, 17.0% vs. 11.1% respectively). CONCLUSION: By adding GM-CSF in culture medium, the quality of embryo, pregnancy and implantation rate could be improved.

Analysis of Androgen Receptor Gene in Korean Patients with Androgen Insensitivity Syndrome / 대한산부인과학회잡지

Androgen insensitivity syndrome (AIS) is a X-linked disorder of sexual differentiation resulting from defective androgen receptor (AR) function. Androgens are secreted by the testes of 46,XY individuals, but there is loss of target organ response to the hormone. The abnormalities of AR are due to defects in the AR gene, and many mutations causing AIS have been reported since the cloning of AR gene. In this study, we analyzed the AR genes in twelve Korean patients with androgen insensitivity syndrome: 9 patients with complete AIS and 3 patients with partial AIS DNAs were isolated from patients with AIS, and the coding region of AR gene was amplified by a polymerase chain reaction using 7 pairs of primers (exon B-H). Sequence analysis of the AR gene was performed using direct sequencing and single strand conformational polymorphism (SSCP). The AR gene mutations were identified in 7 out of 12 patients: 6 of 9 patients with complete AIS, and one of 3 patients with partial AIS. Mutations found were as follows: the point mutation (ATT->ACT) at position 680 of exon D, point mutation (TGG->TGC) at position 751 of exon E, point mutation (CAA->TAA) at position 792 of exon F, point mutations (CGC->TGC, GTG->ATG) at position 855 and 866 of exon G, and the deletion of 13 nucleotides (CGTATCATTGCAT) at position 840 of exon G, respectively. To the best of our knowledge, the point mutations found in exon D, exon E, and exon F, and the deletion in exon G have not been observed before. SSCP revealed bands with abnormal mobility in 10 out of 12 patients tested. Mutations were found 5 out of these 10 patients. The other two patients showed no abnormal band on SSCP, but showed mutations by direct sequencing. In conclusion, we have demonstrated the AR gene mutations, including three novel mutations, in Korean patients with AIS, and these abnormalities might be related to the pathogenesis of androgen insensitivity syndrome.

Expression of leptin receptor (Ob-R) in human atherosclerotic lesions: potential role in intimal neovascularization

Neovascularization of the adventitial vasa vasorum with extension into the intima of atherosclerotic lesions is frequently observed, but its pathophysiological significance is still subject to debate. Recently, leptin, the product of the Ob gene, was identified. Leptin, via activation of the endothelial receptor (Ob-R), generates a growth signal involving a tyrosine kinase-dependent intracellular pathway and promotes angiogenic processes. We hypothesized that a high concentration of leptin within vasa vasorum and plaque itself, may influence inflammatory and vascular neovascularization coupling with functional upregulation of the vascular endothelial growth factor (VEGF). Microscopic computerized tomography was utilized for the spatial distribution of vasa vasorum and intimal neovascularization from atherosclerotic human coronary arteries. Atherosclerotic coronary arteries showed a dense plexus of microvessels in the adventitia and plaque itself. Microscopic analysis from human atherosclerotic aortas revealed an increase in the intimal thickness with neovascularization. The immunoreactivity for Ob-R, VEGF and matrix metalloproteinase (MMP) increased in atherosclerotic plaque, predominantly in the endothelial lining of the intimal neovessel and macrophages/foam cells. Our observation of a prominent colocalization between Ob-R, VEGF and MMP supports this hypothesis and these factors participate in the neovascularization of atherosclerotic lesions. The present study is the first report on vascular tissue and it opens a promising perspective concerning future investigations of leptin-dependent modulation of atherogenesis and vascular neovascularization under pathophysiolgical conditions.

Coexpression of cyclooxygenase-2 and matrix metalloproteinases in human aortic atherosclerotic lesions

Inflammation appears to have a major role in the development of atherosclerosis. Cyclooxygenase-2 (COX-2) is involved in the inflammatory response via the generation of prostanoids that, in turn, are involved in the production of matrix metalloproteinases (MMPs). This study aimed to investigate atherosclerosis in human aortas for in situ tissue distribution of COX-2, MMPs including MMP-9 and membrane type 1 MMP (MT1-MMP), and tissue inhibitor of metalloproteinase-2 (TIMP-2). Immunohistochemical studies were performed on atherosclerotic lesions of aortas from patients with aortic aneurysms (n = 4) and dissections (n = 3) by using antibodies to COX-2, MMP-9, MT1-MMP, and TIMP-2. Control tissues were obtained from traumatically dissected aortas (n = 2). All specimens from diseased aortas had atherosclerotic lesions ranging from fatty streak to atheromatous plaques. In control, there was no expression of COX-2, MMP-9, and MT1-MMP in all aortic layers. Immunoreactivity for COX-2 was predominantly noted in macrophages and smooth muscle cells (SMCs) of the intima including atherosclerotic plaque itself and the medial layer of the plaque base, as well as in SMCs and endothelial lining of the vasa vasorum in the adventitia. Immunoreactivity for MMP-9 and MT1-MMP was found in the same distribution as that of COX-2. Additionally, the expression of TIMP-2 increased in relation to MMP-9 expression. This study demonstrates that COX-2 is coexpressed with MMP-9 and MT1-MMP, not only by macrophages and SMCs in atherosclerotic lesions, but also in endothelial lining of the vasa vasorum of human aortas. Thus, vascular inflammatory reactions may influence extracellular matrix remodeling by coactivation of MMPs in the development of atherosclerosis and, in turn, the progression of disease.

Effect of Leptin on the Steroidogenesis of Cultured Human Granulosa Cells / 대한불임학회지

OBJECTIVE: To elucidate the location of leptin and receptors of ovary specimens obtained from patients undergoing hysterectomy by immunohistochemical staining and to determine the effect of leptin on the steroidogenesis of cultured granulosa cells. METHOD: In the culturing process of the granulosa cells, FSH (1 IU/ml)and leptin (50 ng/ml), IGF-I (50 ng/ml) was administered to each study group (Group I: FSH; Group II: FSH, leptin; Group III: FSH, IGF-I, leptin), and the levels of estradiol, progesterone, androstenedione in the culture media was measured by radioimmunoassay. Statistical analysis was conducted by one-way ANOVA with Scheffe test. RESULTS: The results showed that leptin and leptin receptors were both found to be strongly stained in granulosa and theca cells, and also in some interstitial cells. Leptin receptors were also observed in cultured granulosa cells. While there was no statistically significant difference in the androstnedione concentrations between the groups, estradiol concentrations was significantly decreased in Group IV (2202.0+/-151.14 pg/ml) compared to Group III (2859.0+/-122.6 pg/ml), and progesterone concentrations were also significantly decreased in Group II(4696.3+/-190.6 ng/ml) and Group IV (4517+/-206.78 ng/ml) compared to Group III(5546.0+/-179.5 ng/ml). CONCLUSTION: The study result of this study suggest that leptin is directly involved in the regulation of ovarian functions, in particular steroidogenesis.

Expression pattern of progesterone receptor, integrinbeta(3), cyclooxygenase-2 ( COX-2 ) in human endometrium of patient with the disease related implantation / 대한산부인과학회잡지

OBJECTIVE: We compared the expression pattern of progesterone receptor, integrin 3, cyclooxygenase-2 (COX-2) in in-phased endomerium of patient with the disease related implantation and control group, and tried to confirm the clinical efficacy of the immunohistochemical markers for discrimination of occult uterine receptivity defect in in-phase endometrium. STUDY DESIGN: Endometrial tissues were obtained from 60 women with normal (group 1; n = 20), uterine synechiae (group 2; n = 15), and endometriosis (group 3; n = 25). On 7 ~ 8 days after ovulation (POD 7 ~ 8), sex hormone levels were measured and immunohistochemical staining of PR, integrin 3, and COX-2 expression were performed. RESULTS: PR was decreased in the group 2 and increased in the group 3 comparing with the group 1. integrin 3 expression was significantly decreased in the group 2 and 3. COX-2 expression was significantly decreased in the group 2. But, in the group 3, COX-2 expression was slightly increased in glandular epithelial cells, and significantly increased in stromal cells. CONCLUSIONS: In-phase biopsies from patients with endometriosis and uterine synechiae showed different expression pattern of integrin 3, COX-2, and PR compared to the control. The aberrant expression of immunohistochemical markers be associated with occult uterine receptivity defect and produce the useful diagnostic method.

A Case Report of Complete Androgen Insensitivity Syndrome / 대한산부인과학회잡지

The androgen insensitivity syndrome is a heterogeneous disorder with a wide spectrum of phenotypic abnormalities, ranging from complete female to ambiguous forms that more closely resembles males. Mutations of the androgen receptor gene are responsible for a variable degree of impaired androgen action. The complete androgen insensitivity syndrome is characterized by normal female external appearance in spite of the normal male karyotype 46XY with testes and normal testosterone production and metabolism. This is transmitted by X-linked recessive manner. Wolffian duct does not develop. However, m llerian development does not occur in presence of antim llerian hormone activity. Recently we experienced a case of complete androgen insenditirity syndrome. We reported a case with concerned literatures.

High Cyclooxygenase - 2 Expression in Stage IB Cervical Cancer with Lymph Node Metastasis or Parametrial Invasion / 대한산부인과학회잡지

OBJECTIVE: the enzymes cyclooxygenase(COX)-1 and -2 are necessary for the synthesis of prostaglandins. COX-2 is usually absent in normal cells and is upregulated and expressed as a product of the "immediate early" gene during inflammatory processes. In previous studies, the expression of COX-2 has been shown to be induced by prointlammatory cytockines, and suggestions have been made that overexpression of COX-2 supresses apoptosis and is directly related to tumor growth. We the authors have attempted to determine a relationship between the tumor invasion and metastasis of uterine cervical cancer and COX and apoptosis by comparing the protein expression of apoptosis and COX-I and COX-2 in tumor tissues confirmed with cytokeratin, and therefe determine the clinicopathologic risk factors. MATERIALS AND METHODS: The subjects were 18 patients who were FIGO stage IB uterine cervical cancer patients who underwent surgery at the Ajou University Medical Center. The 18 cases were comprised of 12 cases of squamous cell carcinoma, 3 cases each of adenocarcinoma and adenosquamous carcinoma. There were 9 cases with lymph node or prarametrial involvement and 13 cases with lymphvascular space involvement. All tissues obtained from the cases were subject to immunohistochemical staining for COX-1, -2 and TUNEL method for apoptosis detection, and the following results were obtained. RESULTS: Tumor tissues confirmed by cytokeratin wae separated into tumor surface, tumor stroma, and invasion site portions, and in which increased apoptosis was observed in the tumor surface and tumor stmma, but not in the invasion sites. COX-2 expression was observed in all tumor tissues, which was especially strong in the tumor invasion site. Therefore, it is suggested that COX-2 expression may supress cell apoptosis at the site of tumor invasion. When COX-2 expression was investigated when the cases were divided into groups with regard to the presence or absence of lymph node or parametrial involvement, there was statistically significant (Mann-Whitney U test) COX-2 expression seen microscopically in the tumor stroma (p-value=0.028) and tumor invasion site (p-value=0.040) compared to the tumor surface (p-value=0.499). In other words, in surgically treated stage IB cervical cancer patients, COX-2 was significantly expressed when lymph node or parametrial involvement was present. CONCLUSIONS: These results suggest that the expression of COX-2 in stage IB cervical cancer patients may downregulate apoptosic processes and thus enhances tumor invasion and metastasis.

The Effect of Overweight and Pregnancy on Endocrinologic Features and Glucose Metabolism in Patients with Polycystic Ovarian Syndrome / 대한산부인과학회잡지

OBJECTIVE: The purpose of this study attempts to determine the endocrinologic characteristics and changes in glucose metabolism before/during pregnancy according to different body weights in women with Polycystic ovarian syndrome (PCOS). METHODS: 94 women dia with PCOS were evaluated through measuring serum hormone levels and oral glucose tolerance tests preconceptionally and gestationally. RESULTS: In patients who were of normal weight showed significantly increased serum LH levels compared to those who were overweight (12.8+/-0.9 Vs. 7.1+/-0.8 mIU/ml, p= 0.000), and the serum levels of insulin was increased significantly in the overweight group (7.1+/-0.7 Vs. 15.2+/-2.8 ulU/ml, p= 0.000). the IGFBP-I (32.8+/-10.6 Vs. 8.3+/-2.5 ng/ml, p=0.034) and SHBG (55.8+/-4.2 Vs. 37.1+/-3.1 nmol/ml, p= 0.001) were significantly lower in the ovnweight group. The oral glucose tolerance test before/after pregnancy showed increased frequency of abnormal glucose metabolism, in both of the non-obese group (38.8%, 26,9%) and the obese group (64.2%, 53.9%) compared with normal population. CONCLUSION: It is thought that in the normal weight group with polycystic ovarian syndrome androgen production is stimulated in the theca cells by abnormally high LH secretion, while in the overweight group the hyperinsulinemia state which decreases the SHBG and IGFBP-I, lead to increase biologically active hee androgens and IGF-I and increase insulin binding to its receptor. And during/before pregnancy, women with PCOS showed that incidence of abnormal glucose metabolism was significantly increased in both of non-obese and obese groups.

Expression pettern of Sex Hormone Receptors , Integrins , Cyclooxygenases ( COX ) in Human Endometrium During Menstrual Cycle / 대한산부인과학회잡지

STUDY DESIGN: Tissues were obtained from the endometrium of the posterior hmdus in 42 women (proliferative phase-25 cases, secretory phase-17 cases) with normal menstrual cycles(28-32days interval). The specimens were stained with H&E stain and classified according to the method by Noyes et al(1950) into early proliferative phase(-5~-10days from ovulation), late proliferative phase(-4days~ovulation), early secretory phase (ovulation ~5days), mid-secretory phase(6~10days from ovulation), and late secretory phase(11-14days from ovulation). Immunohistochemical staining of integrin a1, a4, b3, COX-1,-2, ER, PR expression was performed. RESULT: The expression of ER was high in the proliferative phase and low during the secretory phase. The late proliferative phase showed the highest intensity(p<0.05). On the other hand, the expression of PR in stromal cells was relatively uniform during the entire menstrual cycle. However, in epithelial cells, there was a characteristic peak intensity in the late proliferative phase and low intensity in the secretory phase.The expression of integrin a1, a4, b3 in epithelial cells showed no particular pattern in the proliferative phase but showed specific findings in the secretory phase. In the epithelial cells, the intensity of a I staining was increased after the early proliferative phase and sustained during the whole secretory phase(p<0.05), a4 was increased in the early and mid-secretory phases, b3 was increased in the mid-secretory phase to late secretory phase. But the strumal cells were weakly expressed in the whole menstrual cycle but showed no particular pattern, In glandular epithelial cells and stromal cells, COX-1 showed a cyclic pattem according to menstrual cycle; it was strongly expressed in the mid-secretory phase in glandular epithelial cells and mid-secretory and menstrual phase in stromal cells(p<0.05). But in luminal epithelial cells, COX-1 was expressed in the entire menstrual cycle but had no particular pattern. In glandular epithelial cells, stromal cells, and luminal epithelial cells, COX-2 was not expressed during the secretory phase but strongly expressed in the mid-secretory phase(p<0.05). CONCLUSION: The expression of a1, a4, b3, and COX-2 showed as stonng staining during the mid secretory phase which represents the implantation period. The PR expression in epitbelial cells was decreased during same period. These characteristic findings will provide helpful information far histological methods of endormetrial dating and will be useful in the measurement of endometrial maturation during the implantation period.

3-Dimensional Culture System of Endometrial Cells for Studying the Human Implantation Mechanism / 대한불임학회지

In order to study the implantation mechanism various methods for culture of endometrial cells in vitro have been attempted. However, a disadvantage is that primary cultures of stromal and epithelial cells do not have the ability to differentiate, and therefore cannot be reproduced in the same manner as in vivo endometrium. The object of this study is to establish a three dimensional culture of endometrial cells which are both morphologically and functionally identical to in vivo endometrium. Endometrial tissues obtained after hysterectomies were cut into thin slices and treated with collagenase and trypsin-EDTA. The stromal cells and the epithelial cells were separated by centrifugation and cultured for 24 hours in DMEM media containing 10% FCS, 100 nM progesterone, and 1 nM estradiol. The cultured stromal cells were mixed with collagen gel and solidified, after which it was covered with matrigel. Epithelial cells were inoculated on the top and then cultured for 3 days. The three dimensionally cultured endometrial cells were stained for integrin alphal, alpha4, beta3, and cyclooxygenase-1, -2 by immunohistochemistry, which all showed strong expression. The cultured epithelial cells showed the formation of microvilli, tight junctions and pinopodes by electron microscopy. Studies are currently under way utilizing this three dimensional culture model to ascertain the interaction between the embryo and human endometrial cells at the time of implantation, and it is thought that further studied into a new culture environment which would allow longer periods of culture will be necessary.

Effects of GnRH Agonist Used for Ovarian Hyperstimulation in Human IVF-ET on the Apoptosis of Preovulatory Follicular Cells / 대한불임학회지

There have been many reports to date regarding the role of GnRH as a local regulatory factor of ovarian function as studies of human and rat ovaries revealed GnRH and its receptor. In recent studies it has been shown that GnRH directly causes apoptosis in the granulosa cells of the rat ovary, and such results leads to the suggestion that the use of GnRH agonist for more stable long term ovarian hyperstimulation in human IVF-ET programs causes granulosa cell apoptosis which may lead to follicular atresia. Therefore this study attempts to determine if granulosa-luteal cell apoptosis occurs in patients during IVF-ET programs in which GnRH agonist is employed for ovarian hyperstimulation. The quality of oocyte-cumulus complexes obtained during ovum pickup procedures were assessed morphologically and then the fertilization rate and developmental rate was determined. Apoptotic cells among the granulosa-luteal cells obtained during the same procedure were observed after staining with Hematoxylin-rosin. The fragmentation degree of DNA extracted from granulosa-luteal cells was determined and comparatively analyzed. There was no difference in the average age of the patients, the number of oocytes retrieved, and fertilization and developmental rates between the FSH/hMG group and GnRH-long group. There was also no difference in the apoptosis rate and pyknosis rate in the granulosa-luteal cells between the two groups. However, when the oocyte-cumulus complexes were morphoogically divided into the healthy group and atretic group without regard for the method of hyperstimulation, the results showed that the number of oocytes obtained averaged 11.09+/-8.75 and 10.33+/-4.53 per cycle, respectively, showing no significant difference, but the fertilization rate (77.05%, 56.99%, respectively, p<0.01) and developmental ,ate (65.96%, 41.51%, respectively, p<0.01) was significantly increased in the healthy group when compared to the atretic group. The degree of apoptosis in the granulosa-luteal cells showed that in the healthy group it was 2.25% which was not significantly different from the atretic group (2.77%), but the pyknosis rate in the atretic group (27.81%) was significantly higher compared to the healthy group (11.35%, p<0.01). The quantity of DNA fragmentation in the FSH/hMG group was 32.22%, while in the GnRH-long group it was 34.27%, showing no significant difference. On the other hand the degree of DNA fragmentation was 39.05% and 11.83% in the healthy group and atretic group, respectively, showing significantly higher increase in the atretic group (p<0.01). The above results suggest that death of granulosa-luteal cells according to the state of the oocyte-cumulus complex is more related to pyknosis rather than apoptosis. Also, the GnRH agonist used in ovarian hyperstimulation does not seem to directly affect the apoptosis of retrieved oocytes and granulosa-luteal cells, and which is thought to be due to the suppression of the apoptogenic effect of GnRH agonist as a result of the high doses of FSH administered.