Extracellular matrix component is degraded by enzymes of thematrix metalloproteinases(MMPs). MMPs are produced by both hemopoietic and structural cells. Increased activity of MMP-3 in periodontium is strongly associated with inflammatory periodontal disease. The purpose of the present study was to determine the effect of tetracycline analogues on the activity of MMP-3. Tetracycline-HCl, doxycycline-HCl, and minocycline-HCl were applied to huamn gingival fibroblasts at various concentrations of 10, 25, 50, 100, 200microgram/ml, and 1 hour later IL-1beta of 25ng/ml was added. After incubation for 24 hours the cells were reacted by enzyme-linked immunosorbent assay using proMMP-3 ELISA kit. The optical density was measured by microwell plate reader at 450nm. The relative activity of MMP-3 was calculated as the percentage of the optical density of each experimental group to that of the control. The difference of the optical density and the relative activity of MMP-3 between the experimental groups and the control wasstatistically analyzed by one way ANOVA. The results were as follows: 1. Tetracycline-HCl showed the tendency to inhibit the activity of MMP-3 at the concentration lower than 25 microgram/ml, but increased significantly the activity of MMP-3 at the concentration of 200microgram/ml(p<0.05). 2. Doxycycline-HCl inhibited significantly the activity of MMP-3 at the concentration lower than 100microgram/ml, but increased significantly the activity of MMP-3 at the concentration of 200microgram/ml(p<0.05). 3. Minocycline-HCl inhibited the activity of MMP-3 at the concentration in the range of 10 to 200microgram/ml. Within the limit of the present study, the above results suggested that the low concentration of tetracycline analogues could inhibit the activity of MMP-3 induced by IL-1beta in human gingival fibroblasts.
Enzyme-Linked Immunosorbent Assay ; Extracellular Matrix ; Fibroblasts* ; Humans ; Interleukin-1beta ; Matrix Metalloproteinases ; Periodontal Diseases ; Periodontium ; Tetracycline*

OBJECTIVES: To determine whether the body weight, body mass index (BMI), and basal serum level of LH, FSH, testosterone (T), dehydroepiandrosterone sulfate (DHEA-S) are related to the ovarian response to clomiphene citrate (CC) in patients with polycystic ovarian syndrome (PCOS). MATERIALS AND METHOD: From January 1996 to June 1997, total 57 patients with PCOS were enrolled in the present study. Women who had other infertility factors were excluded from our study. The ovulation induction using CC was used in all patients. The patients were grouped into 50 mg group, 100 mg group, and 150 mg group according to their daily CC dose. The patients were also grouped to ovulatory and non-ovulatory group. The body weight, BMI, arid basal serum level of LH, FSH, T, DHEA-S were measured in all patients on the 2nd or 3rd day of the menstrual cycle. Results were analysed with Student's t-test and Fisher's exact test. RESULTS: The body weight and BMI of the nonovulating group were significantly higher than those of the ovulating group in all groups (50, 100, 150 mg of CC). However, there were no significant differences of the level of LH and FSH between ovulating and nonovulating groups in all CC groups (50, 100, 150 mg). The level of T of nonovulating group was significantly higher in 50 and 100 mg of CC groups, but not in 150 mg group. The level of DHEA-S of the non-ovulating group is significantly higher in 50 mg group, but not in 100 and 150 mg groups. CONCLUSION: The body weight and BMI could be useful predictors of ovarian response to CC in patients with PCOS, and basal T and DHEA-S also might be useful in cases of low-dose CC treatment.
Body Mass Index ; Body Weight ; Clomiphene* ; Dehydroepiandrosterone Sulfate ; Female ; Humans ; Infertility ; Menstrual Cycle ; Ovulation Induction ; Polycystic Ovary Syndrome* ; Testosterone

Adult ; Connective Tissue ; Extremities ; Head ; Histiocytoma, Malignant Fibrous ; Humans ; Maxilla ; Neck ; Radiotherapy ; Sarcoma

This study was performed to investigate the influence of epidermal growth factor (EGF) on preimplantation development, implantation, and expression of epidermal growth factor receptor (EGFR) in mouse embryos. Riverse transcription-polymerase chain reaction (RT-PCR) has been used to examine the presence of transcripts. Following reverse transcription, strategically designed nested primers, optimised for specificity, were used for amplification from the cDNA equivalent of a single embryo. The products were then verified by restriction enzyme digestion and sequence analysis. Eight-cell stage mouse embryos were cultured for 48hrs with EGF at concentrations of 0.1, 1.0, 10 and 100 ng/ml. Embryos not treated with EGF were served as control. The percentages of embryos which developed to the expanded, hatched blastocyst stage and in vitro implantation at 48hrs were determined. The percentages of fully expanded murine blastocysts at 48hrs in all EGF treated group were not significantly different from the control. The percentages of hatched blastocysts were significantly higher in EGF treatment group at 0.1ng/ml (90.7%), 10 ng/ml (89.3%) compared to the control (82.1%; p < 0.05, p < 0.05). The percentages of implanted blastocyst in vitro were significantly higher following incubation with EGF at concentrations of O.lng/ml (38.1%; p < 0.05), 1.0ng/ml (33.3%; p < 0.05), 10ng/ml (22.2%; p < 0.05) compared to the control (10.7%). Embryo development and implantation in vitro were not significantly inhibited or enhanced in cultures supplemented with 100ng/ml EGF compared to the control. The mRNA concentration of EGFR in embryos treated with 0.1ng/ml of EGF were significantly higher than those of the control and other EGF treatment groups. The implantation rate and mRNA concentration of EGFR in embryos treated with 0.1ng/ml of EGF group were significantly higher than those of other treatd groups. In conclusion, EGF may have a stimulatory role in embryonic development, implantation and expression of EGFR in embryo itself with concentration-specific manner. These results suggest that EGF may act directly on the mouse embryo and favor its implantaion, irtespective of the presence ar absence of the endometrium.
Animals ; Blastocyst ; Digestion ; DNA, Complementary ; Embryonic Development ; Embryonic Structures* ; Endometrium ; Epidermal Growth Factor* ; Female ; Mice* ; Pregnancy ; Receptor, Epidermal Growth Factor ; Reverse Transcription ; Rivers ; RNA, Messenger ; Sensitivity and Specificity ; Sequence Analysis

With the advent of intracytoplasmic sperm injection (ICSI), the fertiliution and pregnancy have been reported even in complete asthenozoospermia. However, the results of ICSI in men with totally immotile sperm was still disappointing. The reasons for the total lack of sperm movement are not yet determined. The immotility of ejaculated spermatozoa is probably caused by sperm degeneration during epididymal transport, therefore higher viable spermatozoa can be obtained by testicular sperm recovery in some cases with low viability and total lack of movement of ejaculated spermatozoa. Recourse to testicular spermatozoa for ICSI is thus an alternative treatment possibility in this kind of pathology. This clinical study was performed to investigate the efficacy of ICSI employing testicular and ejaculated spermatozoa. From December 1995 to March 1998, 35 couples with totally immotile spermatozoa were included in the study. In 14 patients, the ejaculated spermatozoa were used whereas in 21 patients the spermatozoa were recovered from the testis. There were no significant differences in the fertilization and cleavage rates between the testicular sperm group with 62.6%, 52.7% and the ejaculated sperm group with 56.1%, 74.3%. Two pregnancies were achieved, one in the testicular spnm poup and the other in the ejaculated group. A healthy child was delivered at term in the ejaculated sperm group, but a clinical abortion occurred in a pregnancy in the testicular sperm group. In conclusion, it was suggested that ICSI using ejaculated sperm might be a primary treatment in cases with totally immotile spermatozoa and, if failed, testicular sperm could be used as an alternative mode of heatment.
Asthenozoospermia ; Child ; Family Characteristics ; Fertilization ; Humans ; Male ; Pathology ; Pregnancy ; Pregnancy Rate ; Sperm Injections, Intracytoplasmic* ; Spermatozoa* ; Testis

It was reported that the etiologies of recurrent spontaneous abortion are immunologic factors, endocrinologic problems, anatomical abnormalities, genetic abnormalities, infection, and unexplained factors. Among those etiologic factors, genetic abnormalities occur in about 5% of the couples who experience recurrent spontaneous abortions, and most common parental chromosomal abnormality contributing to recurrent abortion is balanced translocation. The advent of in vitro fertilization (IVF), the development of skills associated with the handling of human embryo, and an explosion of knowledge in molecular biology have opened the possibility of early diagnosis of genetic disease in preimplantation embryos. Therefore preimplantation genetic diagnosis (PGD) is indicated for couples, infertile or not, at risk of transmitting a genetic disease. A case of successful pregnancy and term delivery by PGD using fluorescence in situ hybridization (FISH) technique in patient with recurrent spontaneous abortion due to balanced translocation is presented with brief review of literatures.
Abortion, Habitual ; Abortion, Spontaneous* ; Blastocyst ; Chromosome Aberrations ; Early Diagnosis ; Embryonic Structures ; Explosions ; Family Characteristics ; Female ; Fertilization in Vitro ; Fluorescence ; Humans ; Immunologic Factors ; In Situ Hybridization ; Molecular Biology ; Parents ; Pregnancy* ; Preimplantation Diagnosis* ; Prostaglandins D

It is well known that the clinical test for responsibility of accurate fertilization capacity in male partners is very important to diagnose and treat the infertility. However, it has been reported that the traditional semen analysis cannot accurately predict fertilization and pregnancy potential. The present study was performed to evaluate the acrosomal reaction to ionophore challenge (ARIC) test as a prognostic indicator for fertilization of sperm and oocyte in an in vitro fertilization and embryo transfer (IVF-ET) program. From March 1996 to Februry 1997, 30 couples undergoing IVF program were allocated to this study group. All female partners in the study group were 35 years old or less and their serum level of basal follicle stimulating hormone (FSH) and estradiol (E2) were normal. All the male partners have normal parameters of semen analysis. The ARIC tests were performed on the day of ovum pick up and in vitro insemination in all the male partners. The controlled ovarian hyperstimulation (COH) using luteal long protocol of gonadotropin releasing hormone (GnRH) agonist was used in all couples for IVF-ET. The acrosomal reaction with 10microliter of 10% DMSO was induced spontaneously in 10.1+/-9.8%, and acrosomal reaction with calcium ionophore A 23187 was induced in 27.4+/-18.1%, and the ARIC value was 17.4+/-16.2%. There were no significant correlation between the ARIC value and the fertilization rate (r2=0.044, p=0.268). There were also no significant correlation between the ARIC value and the percentage of the grade I, II embryos (r2=0.046, p=0.261). On the basis of above results, it was suggested that ARIC test might not be a useful prognostic indicator for fertilization in IVF-ET in male partners with normal parameters of conventional semen analysis. We guessed that IVF-ET could be performed to the patients primarily without universal appilcation of ARIC test to all male partenrs, and if fertilization failure occurs, the microassisted fertilization (MAF) such as intracytoplsmic sperm injection (ICSI) might be used as an alternative mode of treatment with acceptable success rate.
Acrosome Reaction* ; Acrosome* ; Adult ; Calcimycin ; Calcium ; Dimethyl Sulfoxide ; Embryo Transfer ; Embryonic Structures ; Estradiol ; Family Characteristics ; Female ; Fertilization ; Fertilization in Vitro* ; Follicle Stimulating Hormone ; Gonadotropin-Releasing Hormone ; Humans ; Infertility ; Insemination ; Male ; Oocytes ; Ovum ; Pregnancy ; Semen Analysis ; Spermatozoa

OBJECTIVE: To compare the pregnancy outcomes of transvaginal multifetal pregnancy reduction (MFPR) according to the gestational period when the procedure was performed METHODS: From January l995 to February 1998, total 27 patients with multiple pregnancy were included in this study. The patients were grouped to early MFPR group (<8 weeks, n=16) and delayed MFPR group (>8 weeks, n=ll) according to the gestational age that MFPRs were performed. All MFPRs were performed by transvaginal sonography-guided fetal aspiration or mechanical trauma. The complete pregnancy loss rate before 24 weeks of gestation, spontaneous loss of embryo, procedure-related complication, gestational age at delivery, and pregnancy complication were compared between the two groups. Statistical analysis of data was performed using Students t-test and Fishers exact test as appropriate. Statistical significance was defined as p<0.05. RESULTS: There was no significant difference in the complete pregnancy loss rate between the early MFPR group (6.3%) and the delayed MFPR group (27.3%). The incidence of partial spontaneous loss of embryo in the two groups were not differed significantly (6.3% vs. 18.2%). The procedure-related complication of the delayed MFPR group (36.4%) seemed to be higher than that of the early MFPR group (6.3%), however there was no statistical difference (p=0, 07). Especially, all 3 patients in whom the MFPR was performed after 10 weeks suffered from the procedure-related complication. The mean gestational age at delivery of the two groups were not differed significantly (36.3+2.8 weeks vs. 37.0+1.3 weeks). There was also no significant difference in the mean birth weights of the two groups (2378.8+563.7 gm vs. 2427.1+436.2 gm). CONCLUSION: Although there was no statistically significant difference, the early transvaginal MFPR might be a safe and useful method without significant adverse complications compared to the delayed MFPR.
Birth Weight ; Embryonic Structures ; Female ; Gestational Age ; Humans ; Incidence ; Pregnancy ; Pregnancy Complications ; Pregnancy Outcome ; Pregnancy Reduction, Multifetal* ; Pregnancy, Multiple

OBJECTIVES: To investigate the patterns of biochemical bone markers, such as urinary deoxypyridinoline (DPD), N-telopeptide of type I collagen (NTX), and serum osteocalcin (OC), bone-specific alkaline pbosphatase (BSAP) in postmenopansal women with hormone replacement therapy (HRT). Materials and METHOD: From July 1997 to January 1998, total 239 postmenopausal women were emolled in the present study, and 198 healthy premenopausal women with regular menstruation were served as control. The postmenopausal women were pouped into the HRT group and the non-HRT group. The women in the HRT poup have received estrogen with continuous or cyclic progestin therapy far more tban 6 months. The biochemical bone markers of all women were assayed. Results were analysed with Students t-test. RESULTS: The urinary DPD of the non-HRT group was sigaificantly higher than both the HRT poup and the premenopausal group(5.51 +/- 2.47 vs. 3.36 +/- 1.02 and 4.01 +/- 3.86 nM/mM, p < 0.05, repectively). The urinary NTX of the non-HRT group was also higher in comparison to the HRT group and the premenopausal group(48.71 +/- 11.54 vs. 33.70 +/- 17.43 and 33.70 +/- 17.43 nM BCE/mmol, p < 0.05, repectively). However, there were no significant differences in the concentrations of serum BSAP and OC among the three poups. CONCLUSION: The urinary DPD and NTX were more sensitive indicators of bone metabolism tban serum BSAP and OC in postmenopausal women undergoing HRT.
Collagen Type I ; Estrogens ; Female ; Hormone Replacement Therapy* ; Humans ; Menstruation ; Metabolism ; Osteocalcin

OBJECTIVE: To determine if basal serom follicle stimulating harmone (FSH) level could be a prognostic factor of the clinical outcome in in vitro fertilization and embryo transfer (IVF-ET) with intracytoplasmic sperm injection (ICSI) in the couples with male factor infertility. Materials and METHOD: From December 1995 to March 1998, total 118 patients with male factor infertility were included in this study. Patients were allocated to the low basal FSH group (>8.5 mIU/ml) and the high basal FSH group (>8.5 mlU/ml). The basal levels of FSH were measured in the 3rd day of menstrual cycle preceding ovarian stimulation cycle in total IVF cycles by immunoradiometric assay (IRMA). Statistical analysis was performed using Student's t-test, Fishers exact test, and x2 test as appropriate. Statistical significance was defined as p < 0.05. RESULTS: The total dose of exogeneous gonadotropin required in the patients of the high basal FSH group was significantly higher than that of the patients with the low basal FSH poup (p < 0.05). The numbers of retrieved oocytes, oocytes with grade I, II, fertilized oocytes, cleaved oocytes, embryos with grade I, II, and transferred embryos were significantly higher in the low basal FSH group (p < 0.05), The clinical pregnancy rate per cycle in the low basal FSH gmup (15.7%) seemed to be higher than that in the high basal FSH poup (3.4%) (p 0.08), however, there was no statistically significant difference between the two groups. CONCLUSION: These results suggested that the basal FSH levels could be predictive of pregnancy outcome and the results of ovarian stimulation in IVF-ET using ICSL.
Embryo Transfer ; Embryonic Structures ; Family Characteristics ; Female ; Fertilization in Vitro ; Gonadotropins ; Humans ; Immunoradiometric Assay ; Infertility ; Male ; Menstrual Cycle ; Oocytes ; Ovulation Induction ; Pregnancy ; Pregnancy Outcome ; Pregnancy Rate ; Sperm Injections, Intracytoplasmic*