The use of hormonal stimulation in human in vitro fertilization and embryo transfer (IVF-ET) leads to increased production of embryos for ET. So to avoid high pregnancies and to allow conception in future, unstimulated cycles, cryopreservation of spare embryos is desirable. One of the improvement of cryopreservation methods is vitrification. We cryopreserved mouse day 3 embryos by vitrification using the three different vitrification solution (EFS40, VSll and VS3a). EFS40 solution is consisted of 40% (v/v) ethylene glycol, Ficol170 30% (w/v) and 0.5M sucrose and VSll is 6.0M ethylene glycol and 1.8M glycerol. And VS3a is 6.5M glycerol and 6% (w/v) BSA (bovine serum albumin). First we tested the toxicity of three vitrification solution by exposure to these solution during 3 min. After washing by thawing solution, the survival rates of each groups are 95.5%, 90.9% and 84.4% (EFS40, VS11 and VS3a). High percentages of them developed to expanded blastocyst and hatching embryos in culture 48hrs 94.2%, 97.7%, 100% and 97.4% (no treatment group, EFS40, VS11 and VS3a). So there is no significant differences among the each group. Second, after thawing of vitirfied embryos, the survival rates of each groups are 96.8% (slow freeze), 94.1% (EFS40), 85.5% (VS11) and 80.0% (VS3a, P vs. no freeze or EFS40 is 0.01). Vitrified embryos exhibited a high rate of development in vitro after 48hrs culture. The percentages of each group to blastocyst and hatching embryos are 88.7% (no freeze), 91.8% (slow freeze), 93.4% (EFS40), 87.7% (VS11) and 73.0% (VS3a, P vs. other group is 0.01). The results suggest that there is no significant differences in exposure of various vitrification solution and day 3 mouse embryos can be vitrified in solution EFS40 and VS11 by simple procedure.
Animals ; Blastocyst ; Cryopreservation* ; Embryo Transfer ; Embryonic Structures* ; Ethylene Glycol ; Fertilization ; Fertilization in Vitro ; Glycerol ; Humans ; Mice* ; Pregnancy ; Sucrose ; Survival Rate ; Vitrification*

The purpose of this study was to investigate effects of the developed nutrition education program focused on individual daily energy requirements and food exchange units using Food Exchange System for diabetes mellitus at a community health center. Developed the nutrition education program, four weeks' nutrition education including provided twice individual meal as diet therapy (2 hour/lesson/week, 4 week), was provided to 20 diabetic elderly (12 male, 8 female, 50-75 yrs): 1st lesson "Introduction: management of diabetes mellitus", 2nd lesson "6 Food groups and sources of 6 food groups", 3rd lesson "Individual daily energy requirements and food exchange units", and 4th lesson "Food choice for diabetes mellitus". For effects' analysis of the developed program, we assessed the changes in anthropometric characteristics; biochemical characteristics and nutrient intakes using 24 hr recall method. Effects of the developed nutrition education program were as follows: weight was significantly decreased, blood urea nitrogen (BUN) and glycosylated hemoglobin (HbA1c) were significantly decreased, and distribution of subjects in BUN and HbA1c was significantly changed. In protein : fat : carbohydrate (PFC) ratio, it was significantly changed from 15.98 : 16.30 : 66.69 to 17.51 : 18.94 : 64.10. In evaluation of nutrient intakes by Dietary Reference Intakes for Koreans (KDRI), protein, fiber, fat, vitamin E, niacin, folic acid, calcium and zinc were shown significantly positive changes in distribution of subjects according to intake level. The index of nutrition quality (INQ), nutrition adequacy ratio (NAR) and mean nutrition adequacy ratio (MAR) were significantly increased. In conclusion, the developed 4 weeks' nutrition education program focused on individual daily energy requirements and food exchange units using Food Exchange System for diabetes mellitus at community health center may improve the symptom of diabetes mellitus.
Aged ; Blood Urea Nitrogen ; Calcium ; Community Health Centers ; Diabetes Mellitus ; Female ; Folic Acid ; Hemoglobin A, Glycosylated ; Humans ; Male ; Meals ; Niacin ; Vitamin E ; Vitamins ; Zinc

Normal and abnormal follicular growth and steroidogenesis depend on gonadotropins as well as intraovarian peptides, which may mediate or potentiate gonadotropin action. Inhibin also affect follicular development and steroidogenesis and may play a role in dominant follicle selection and follicular atresia. Therefore, we studied the differences of serum inhibin, gonadotropin and androgen levels in the women with only the ultrasound findings and no disorder, and polycystic ovary (PCO) with ovulatory disturbance. We prospectively analysed forty-three women with PCO. The diagnosis of PCO was based on typical appearance of the ovaries on TVS. Twelve women with regular menstrual cycle and normal ovarian morphology were selected as control. Basal levels of inhibin, luteinizing hormone(LH), follicle stimulating hormone(FSH), estradiol(E2), testosterone(T), androstenedione(ADD), dehydroepiandrosterone-sulfate(DS), prolactin and TSH in serum were determined. There were significant differences in basal LH levels and LH/FSH ratio between the control and the women with PCO. The basal levels of inhibin and E2 in the oligo-amenorrheic PCO (N=34) were significantly higher than those in the control. There was higher negative correlation between the inhibin and T levels in the oligo-amenorrheic PCO, but, not in the regular cycling PCO. Also, there was higher positive correlation between the LH and T levels in the oligo-amenorrheic PCO, but not in the regular cycling PCO. These data presume that the initial event of PCO is elevated pituitary LH secretion. Elevated levels of LH may down-regulate LH receptors on granulosa cells and also cause hypertrophy of the thecal layer. High level of androgen secreted by the hypertrophied thermal layer may stimulate inhibin secretion from granulosa cells and can be converted to estrogen by extraovarian tissues and could serve to augment pituitary sensitivity to GnRH with a resultant secretion of more LH follicular development and dominant follicle selection resulted in ovulatory disturbance.
Diagnosis ; Estrogens ; Female ; Follicular Atresia ; Gonadotropin-Releasing Hormone ; Gonadotropins ; Granulosa Cells ; Humans ; Hypertrophy ; Inhibins* ; Lutein ; Menstrual Cycle ; Ovary* ; Peptides ; Prolactin ; Prospective Studies ; Receptors, LH ; Ultrasonography

OBJECTIVE: The involvement of IGF system in hyperandrogenism and abnormal follicular development is controversial. This study is to assess whether IGF system contribute to it in the women with polycystic ovary(PCO). METHODS: Baseline serum levels of luteinizing hormone (LH), follicle stimulating hormone (FSH), estradiol (E2), testosterone (T), androstenedione (ADD), prolactin, thyroid stimulating hormone (TSH), free insulin-like growth factor(IGF)-I, free IGF-II, insulin-like growth factor binding protein(IGFBP)-1, and IGFBP-3 were measured in twelve healthy regularly cycling volunteers and forty-two women with PCO then, the changes of baseline serum levels were evaluated after laparoscopic ovarian electrocauterization in nine PCO patients. In addition, the expression pattern of IGF-I and IGF-II was examined in the ovary of control and PCO group. RESULTS: Baseline levels of LH, ADD, free IGF-II, and IGFBP-3 were significantly higher in PCO group. However, there were no significant differences in the levels of free IGF-I and IGFBP-1, although free IGF-I showed decreasing tendency in PCO group. And there was a significant positive correlation between the LH and free IGF-II level in the PCO(P=0.011, r2=0.3899), but not in the control. After ovarian electrocauterization, LH, T, and ADD levels decreased, and free IGF-I and IGFBP-3 level increased. While free IGF-II and IGFBP-1 level showed no significant changes. In the ovary, expression of both IGFs showed similar pattern in normal and PCO ovaries. CONCLUSIONS: The elevated IGFBP-3 level may alter the bioavailability of IGF(s) in the PCO. The change in IGF-I level and resumption of ovulation after electrocauterization, suggest a possible role of IGF system in the impairment of follicular development in the PCO.
Androstenedione ; Biological Availability ; Carrier Proteins* ; Estradiol ; Female ; Follicle Stimulating Hormone ; Humans ; Hyperandrogenism ; Insulin-Like Growth Factor Binding Protein 1 ; Insulin-Like Growth Factor Binding Protein 3 ; Insulin-Like Growth Factor I ; Insulin-Like Growth Factor II ; Luteinizing Hormone ; Ovary* ; Ovulation ; Prolactin ; Somatomedins* ; Testosterone ; Thyrotropin ; Volunteers

Most mammalian embryos implant on the uterine endometrium after hatching fromzona pellucida of the expanded blastocyst and pregnancy takes place. The blastocysts produceand control a variety of prostaglandins which activate a few proteolytic enzymes thatdissolve the zona pellucida of the embryos. In the present study, the goal was to investigatethe indomethacin and aspirin(inhibitors of cyclooxygenase pathway which regulates thepathway from arachidonic acid to prostaglandins), which can affect the hatching and implantationof the mouse embryos by the treatment of the different dose level of indomethacinand aspirin in the culture of mouse embryos.The female and male ICR mice, 6~8 weeks and were used for superovulation andmating and M16 was used as a basic culture medium.The above results can be summarized as following:1. Indomethacin seemed to inhibit the development of the mouse embryos and hatchingprocess because of inhibiting or blocking the activation of hatching related enzymes andhigh dose of indomethacin inhibited implantation.2. Aspirin had no effect on the hatching of the embryos at the dose of 0.16 mg/ml.3. FBS seemed to contain a factor which induced outgrowth of the embryo whereasBSA did not and outgrowth did not take place in the BSA contained medium.4. Blastocysts produced enough prostaglandins F2alpha which was needed for thehatching whereas they needed a factor for implantation which might be produced in theendometrium or exuded from the blood.In conclusion the concentration of indomethacin used in the present study inhibithatching of the blastocysts. This seems to be caused by inhibiting the synthesis of theproteins need for hatching. The factor that induce the outgrowth of the blastocysts doesnot seem to be produced in the blastocysts themselves but seem to be present in the FBS.
Animals ; Arachidonic Acid ; Aspirin ; Blastocyst ; Embryo, Mammalian ; Embryonic Structures ; Endometrium ; Female ; Humans ; Indomethacin ; Male ; Mice* ; Mice, Inbred ICR ; Peptide Hydrolases ; Pregnancy ; Prostaglandin-Endoperoxide Synthases ; Prostaglandins ; Superovulation ; Zona Pellucida

OBJECTIVE: We reported the overcoming effect of Ni2+ on the in vitro 2-cell block of mouse embryos. In this study, we aim to investigate whether Ni2+ should induce intracellular Ca2+ transient in the mouse embryos. MATERIALS AND METHODS: Embryos were collected at post hCG 32hr from the oviduct of the ICR mouse and cultured in M2 medium omitted phenol red. Intracellular Ca2+ was checked by using a confocal laser scanning microscope and fluo-3AM by using various intracellular Ca2+ antagonists. RESULTS: In 1mM Ni2+ treated medium which contained Ca2+(1.71mM), 75.7% of the embryos showed [Ca2+]i transient about 200 sec later. In the Ca2+-free medium, 69.8% of the embryos showed [Ca2+]i transient. In U73122, phospholipaseC(PLC) inhibitor (5uM, 10min) pretreated group, 33.3% of the embryos showed [Ca2+]i transient. Heparine, inositol 1,4,5-triphosphate receptor(IP3R) antagonist preinjected embryos showed no response with 1mM Ni2+. In danthrolene treatment, ryanodine receptor(RyR)-antagonist, 43% embryos showed [Ca2+]i transient but they showed delayed response about 340sec in the presence of Ca2+. CONCLUSIONS: Summing up the above results, Ni2+ seems to induce Ca2+-release from the Ca2+-store even in the Ca2+-free medium. IP3 receptors of the mouse 2-cell embryos might have an essential role for the intracellular Ca2+ increase by Ni2+.
Animals ; Embryonic Structures* ; Heparin ; Inositol 1,4,5-Trisphosphate ; Inositol 1,4,5-Trisphosphate Receptors ; Mice* ; Mice, Inbred ICR ; Oviducts ; Phenolsulfonphthalein ; Ryanodine

BACKGROUND: Basic Fibroblast Growth Factor (bFGF) is known to be closely related to myelofibrosis and hematopoiesis including magakaryopoiesis. The main bone marrow finding in patients with idiopathic thrombocytopenic purpura (ITP) is an increased megakaryopoiesis without myelofibrosis. Purposes of this study are to evaluate the changes in bFGF expression pattern in the bone marrow of patients with ITP and to correlate them with the plasma concentrations of bFGF. METHODS: Paraffin-sections of bone marrow biopsies from 17 cases ITP and 7 cases normal controls, without pathological alterations, were investigated by immunohistochemistry for bFGF and CD68. The plasma levels of bFGF were evaluated by enzyme immunoassay in 7 cases of ITP and controls. RESULTS: The bFGF was strongly expressed in stromal cells and weakly in megakaryocytes in normal controls. The density of the bFGF-expressing stromal cells was decreased in 70% (12/17) of the patients with ITP, compared with none in the other controls. The number of stromal cells in patients with ITP was similar to those in the control groups. The bFGF plasma levels were significantly lower in almost all the ITP patients compared to the control group. CONCLUSIONS: The results indicate that concentrations of bFGF in plasma and bone marrow stromal cells of ITP were decreased. Although the mechanism of low cellular and plasma concentrations of bFGF needs to be elucidated, these findings may complement the serologic and morphological diagnosis of ITP.
Biopsy ; Bone Marrow* ; Complement System Proteins ; Diagnosis ; Fibroblast Growth Factor 2* ; Hematopoiesis ; Humans ; Immunoenzyme Techniques ; Immunohistochemistry ; Megakaryocytes ; Mesenchymal Stromal Cells ; Plasma* ; Primary Myelofibrosis ; Purpura, Thrombocytopenic, Idiopathic* ; Stromal Cells

A total of 6043 surgical cases were performed at S.D.A. Hospital from 1973 to 1979. These were analyzed statistically according to yearly numbers and emergency states, age, sex, anesthetic methods, anesthetic technique, duration of anesthesia, anesthetic agents and mortality. These were all inpatients operated under general anesthesia, and the results are as follows. Results. 1) Yearly numbers and emergency numbers were steadily increasing. 2) Sex ratio was 65%: 35% 2887 male cases(47.7%) and 5156 female cases(52.23%). 3) More than half of the age group was adult(21 to 60 years) 4) Halothane, nitrous oxide and neuromuscular blocker were used with increasing tendency. 5) The anesthetic technique was, in more than 90% of cases, was circle absorption system. 6) General surgery cases were 40% of the total of all departments. 7) In 66.95% of anesthesias the duration was 1 to 3 hours. 8) General anesthesia group was 98.63%. 9) Deathe occurred in 6 cases our of a total of 6043 cases.
Absorption ; Anesthesia* ; Anesthesia, General ; Anesthetics ; Emergencies ; Female ; Halothane ; Humans ; Inpatients ; Male ; Mortality ; Neuromuscular Blockade ; Nitrous Oxide ; Sex Ratio

Doppler sonography has been introduced to evaluate uterine and ovarian diseases. It has been reported to be used to distinguish benign from malignant adnexal masses. The purpose of this study was to assess the blood flow characteristics of adnexal masses and to determine the optimal cut-off points for the Doppler indices obtained, resistance index and pulsatility index, to best differentiate benign from malignant lesions. We performed pulsed Doppler ultrasound in seventy-eight adnexal masses between February 1989 and June 1994. The pulsatility and resistance index were calculated from the waveforms generated from blood flow within the ovary. There were 51 benign and 27 malignant histologically proved ovarian lesions. All women have been operated on. The results were as follows ; Benign tumors and cysts had a significantly higher pulsatility index(mean, 1.23+/-0.74; range, 4.46-3.36) and resistance index(mean, 0.64+/-0.19; range 0.34-1.43) than did malignant tumors(pulsatility index : mean, 0.87+/-0.49; range, 0.31-2.27; resistance index : mean, 0.50+/-17; range, 0.23-0.8). However, some overlap in individual values for benign and malignant lesions was found. In conclusion, our data suggest that high pulsatility and resistance indices may indicate benign adnexal masses ; however, considerable overlap in pulsatility and resistance indices between benign and malignant lesions were noted, and further work will be needed before the validity of these factors is proved.
Female ; Humans ; Ovarian Diseases ; Ovary ; Ultrasonography

A case of traumatic thoracolumbar epidural hemorrhage is presented in a 37-year-old man with ankylosing spondylitis. The patient complained neck and back pain right after the accident but paralegia and difficult urination followed 2 days later. Spine CT showed thoracolumbar epidural mass, which was confirmed to be hemorrhage associated with spine fracture and was solely responsible for paraplegia.
Adult ; Back Pain ; Hematoma, Epidural, Spinal* ; Hemorrhage ; Humans ; Neck ; Paraplegia ; Spine ; Spondylitis, Ankylosing* ; Urination