OBJECTIVE: Evaluation of embryos using early cleavage to 2-cell stage has been proposed, but a critical time-point for selecting embryos is unclear. The aim of the present study is to provide a guideline including critical time-point in the selection of early cleaving embryo for the reduction of multiple pregnancies as well as the increase of pregnancy rate in human IVF. METHODS: This prospective study was performed in 116 cycles from 85 patients who underwent conventional IVF or ICSI at the infertility clinic of Good Moonhwa Hospital from January 2002 to December 2003. Early cleavage (EC) of embryos to 2-cell stage was assessed at 25 h and 27 h postinsemination/microinjection. Embryos that had early cleaved at each time point were designated as EC-1 and EC-2, respectively, while others were designated as non-early cleavage (NEC). RESULTS: At least one early cleavage embryo was observed in 54 (46.6%) for the EC-1 and 84 (72.4%) for the EC-2 of the 116 cycles assessed. Clinical pregnancy rates (PR) were significantly higher in the EC-1 group (66.7%) compared to the EC-2 group (53.6%) or the NEC group (31.2%) (p<0.05). Significant improvement of the pregnancy rate was found when at least two or more embryos were early cleaved at 25 h postinsemination or when the proportion of early cleavage embryo at 25 h postinsemination was higher than 20% (p<0.05). CONCLUSIONS: The critical time-point for the selection of early cleavage embryos with high implantation potential is more effective in 25 h postinsemination/microinjection compared to 27 h. The proportion as well as number of early cleavage embryos is also an important factor for the prediction of pregnancy outcome and the chance of multiple pregnancies. These results demonstrated that the evaluation of early cleavage embryos to 2-cell stage is an easy, simple, and objective method for the selection of good quality embryos suitable for embryo transfer.
Embryo Transfer ; Embryonic Structures* ; Female ; Humans* ; Infertility ; Pregnancy Outcome ; Pregnancy Rate* ; Pregnancy* ; Pregnancy, Multiple ; Prospective Studies ; Sperm Injections, Intracytoplasmic

No abstract available.
Endometrial Neoplasms* ; Female

OBJECTIVE: A number of studies to improve in vitro culture conditions have been tried over past ten years by using co-culture system with helper somatic cells. However, the mechanism of coculture is poorly understood. This study was designed to understand the mechanism for the mode of actual action of co-culture system of ICR strain's 1-cell embryos with human oviduct epithelial cells by examining the effect of conditioned medium and contactless coculture using a cell culture insert on the embryo development and by measuring the level of superoxide anion from conditioned medium after co-culture. METHODS: ICR strain's zygote embryos were cultured in medium alone (control), coculture, conditioned medium, or contactless coculture system for 6 days. Conditioned media (CM) were prepared as following 5 groups. All CM were collected after culturing oviduct cells for 2 days. CM-1 was stored at -20degrees C until use, and CM-2 was prepared just before use as a culture medium. CM-3 was cocultured with embryos and retrieved just before use. CM-4 and CM-5 were derives from the microfilteration of CM-2 and CM-3, respectively, using Microcon-10 (10 kDa molecular weight cut-off). The percentage of the embryos developed to hatched blastocyst stage and the level of superoxide anion in supernatant from medium alone culture (control), coculture, and contactless coculture were measured. RESULTS: The rates of embryo development to the hatched blastocyst stage were significantly higher in coculture (43%) than in control (0%) (p<0.05). The CM-1 group had no embryo development since 2-cell embryonic stage, whereas the CM-2, CM-3, CM-4 and CM-5 groups had the improved development to 4 or 8 cell embryo stage, but the similar rate of development to hatched blastocyst compared to control. The effect of coculture on embryo develpment was disappeared in the contactless coculture group. The level of superoxide anion was significantly reduced in coculture group compared to control. CONCLUSION: It is concluded that the present coculture system overcomes the 2-cell block in vitro and improves the embryo development. This beneficial effect may be due to the direct cell-cell contact between embryo and helper cells or the removal of deleterious components from medium rather than the embryotrophic factors.
Animals ; Blastocyst ; Cell Culture Techniques ; Coculture Techniques* ; Culture Media, Conditioned ; Embryonic Development ; Embryonic Structures ; Epithelial Cells ; Female ; Humans ; Molecular Weight ; Oviducts ; Pregnancy ; Superoxides ; T-Lymphocytes, Helper-Inducer ; Zygote

This study was performed to determine the effects of nitric oxide on human sperm cell function. Semen samples were obtained from normal healthy volunteers. Motile spermatozoas collected by swim-up method were incubated up to 24 hours in Ham's F-10 medium supplemented with a various concentration of sodium nitroprusside (nitric oxide releasing agent). Sperm motility, hyperactivation, acrosome reaction rate, and acrosin activity were determined. The results are as follows; 1. 1mM of SNP resulted in a significant decrease in sperm motility (44.8%+/- 8.9%:78.1%+/-6.3%, and hyperactivation (10.4%+/-6.4%:477%+/-9.5%) after incubation for 3 hours compared with the control group (Ham's F-10 alone), but had no effect on acrosome reaction. 2. At 100muM SNP, sperm motility was reduced after incubation for 6 hours (54.8%+/- 3.2%) compared with that of the control group (82.7% +/- 8.9%), but hyperactivation and acrosome reaction were not affected. 3. However, a lower concentration (less than 101M) of SNP had no effect on sperm motility and hyperactivation for 8 hours of incubation but significantly decreased them when incubation periods were increased up to 24 hours compared with the control group. On the other hand, 1muM and l0muM SNP significantly increased the acrosome reaction rate in both acrosomal status (17.3%+/-5.2%,23.5%+/-4.7%, respectively) and acrosin activity (34.3muIU+/- 10.5muIU, 45.6muIU+/-5.6muIU, respectively) as compared with the control group (7.0%+/-4.0%, 9.5muIU+/-3.4muIU). These results indicate that SNP, NO releasing agent, has a dose-dependent effects on the sperm cell function. Therefore it may positively affect the fertilization by promoting acrosomal reaction at a lower concentration (less than 101M).
Acrosin ; Acrosome Reaction ; Fertilization ; Hand ; Healthy Volunteers ; Humans* ; Nitric Oxide* ; Nitroprusside ; Semen ; Sperm Motility ; Spermatozoa*

Purpose: To evaluate the clinical results of a "Four-in-One" procedure for a habitual dislocation of the patella in children with generalized ligamentous laxity and formation failure of the femoral trochlea. Materials and Methods: Five knees in 4 patients were included in this study. The average age of the patients at the time of surgery was 5.9 years and the subjects were followed up for an average of 41.6 months postoperatively. The clinical results were evaluated using the criteria of the Kujala's scoring system as well as a physical examination and radiological findings. Results: During the follow-up period, there were no recurrent dislocations, knee joint pain, limitations of motion or gait disturbances in any of the cases. The mean Kujala score was 96.8. Two cases had complications related to wound healing but they healed eventually. Conclusion: The "Four-in-One" procedure, which include the lateral retinacular release, medial vector augmentation, semitendinosus tenodesis, and patellar tendon transfer is recommended for a habitual dislocation of the patella in children with generalized ligament laxity and formation failure of the femoral trochlea.
Arthralgia ; Child* ; Dislocations ; Follow-Up Studies ; Gait ; Humans ; Knee ; Knee Dislocation ; Ligaments* ; Patella ; Patellar Dislocation* ; Patellar Ligament ; Physical Examination ; Tenodesis ; Wound Healing

OBJECTIVE: Alendronate, a widely used bisphosphonates, acts to inhibit bone resorption by interfering with the activity of osteoclasts. Recently, it has been reported that alendronate also may increase bone proliferation and osteoblastic differentiation. However, little is known about mechanism of the action of alendronate on osteoblast differentiation, especially in transcription level. Inhibitors of DNA binding/ differentiation (Ids) are helix-loop-helix (HLH) transcription factors and play an important role in BMP-induced osteoblast lineage-specific differentiation. Therefore, this study was aimed to investigate the effect of alendronate on osteoblast differentiation and expression of Id-1 and Id-2. METHODS: MC3T3-E1, pre-osteoblast cell line, were treated with alendronate of various concentrations (10(-9) M-10(-4) M) and time periods (24, 48 and 72 hours). And then, the effect of alendronate on osteoblast differentiation was examined by alkaline phosphatase (ALP) activity and RT-PCR for osteoblast differentiation markers such as ALP, type 1 collagen (Col 1), and osteocalcin (OCN). The expressions of Id-1 and Id-2 were measured by RT-PCR. RESULTS: Alendronate treatment increased not only ALP activity, but also expressions of ALP, Col 1, and OCN. Also, alendronate treatment up-regulated the mRNA levels of Id-1 and Id-2 genes. This alendronate-induced osteoblastic differentiation is more effective in lower doses rather than high doses. CONCLUSION: This study shows that the expression of transcription factor Id-1 and Id-2 was increased in a dose-dependent manner during alendronate-induced osteoblast differentiation.
Alendronate ; Alkaline Phosphatase ; Antigens, Differentiation ; Bone Resorption ; Cell Differentiation ; Cell Line ; Collagen Type I ; Delta Sleep-Inducing Peptide ; Diphosphonates ; DNA ; Osteoblasts ; Osteocalcin ; Osteoclasts ; RNA, Messenger ; Transcription Factors

OBJECTIVE: Distal middle cerebral artery (MCA) aneurysms are the least frequent aneurysms of the MCA, and they represent about 1.1 to 5% of all MCA aneurysms. Patients with ruptured distal MCA aneurysms generally have a poor clinical outcome. The purpose of this article is to review the characteristics of distal MCA aneurysms to avoid the complications of microsurgical dissection and clipping of distal MCA aneurysms. METHODS: A total of 1187 patients with ruptured aneurysms were treated at our hospital between January 1997 and May 2008. All patients underwent surgical procedures. Computed tomography (CT) revealed rupture of distal MCA aneurysms in 15 (1.26%) patients. The location of the aneurysm were the M2 (insular) segment in seven patients, the M2-3 junction in three and the M3 (opercular) segment in five. Brain CT images revealed the presence of both subarachnoid hemorrhage (SAH) and intracranial hemorrhage (ICH) in 11 of 15 (77.3%) patients, with a mean ICH volume of 14.5 cc (range : 5 to 32 cc). Rebleeding occurred in 7 out of the 15 (46.7%) patients. RESULTS: All the patients underwent early surgical procedures, including clipping in seven, trapping in two, bypass surgery in four, Guglielmi detachable coil embolization in one and exploratory craniotomy in one patient. The aneurysm had a fusiform appearance in 9 out of 15 cases (60%), and the mean size of the aneurysm was 10.4 mm (range : 2 to 35 mm). Three patients died due to severe brain swelling (20%). CONCLUSION: In this study, distal MCA aneurysms had a relatively fusiform shape as well as high rates of rebleeding and ICH. A good clinical outcome was associated with early surgery for adequately controlling brain swelling and preventing rebleeding.
Aneurysm ; Aneurysm, Ruptured ; Brain ; Brain Edema ; Cerebral Hemorrhage ; Craniotomy ; Humans ; Intracranial Aneurysm ; Intracranial Hemorrhages ; Middle Cerebral Artery ; Rupture ; Subarachnoid Hemorrhage

OBJECTIVE: It has been known that amniotic fluid (AF) is rich source of mesenchymal stem cells (MSCs). Bisphosphonates are widely used in clinical treatment of various metabolic bone diseases and their primary action is the inhibition of osteoclastic bone resorption. However, litter is known about whether bisphosphonates affect the differentiation into osteoblast, especially from AF-derived MSCs (AFMSCs). Therefore, the purpose of this study is to investigate whether these bisphosphonates influence in the process of AFMSCs differentiation into osteoblast. METHODS: AF samples were obtained by second trimester amniocentesis for fetal karyotyping from 6 pregnant women. Cells were treated with various concentration (0, 10(-10), 10(-8), 10(-6) M) of zoledronate and alendronate and analyzed over 21 days of culture. Differentiation into osteoblast was determined by cell staining and RT-PCR for alkaline phosphatase (ALP). RESULTS: It was observed that AFMSCs could differentiate into osteoblast. Alendronate had more potent effect than zoledronate in osteoblastic differentiation. ALP expression was increased with increasing concentration of zoledronate and it was highest in 10(-8) M alendronate. However, no effect of bisphosphonates was found in 14 days of culture. CONCLUSION: This study shows that AFMSCs can be differentiated into osteoblast. The induction of these differentiation following bisphosphonate treatment was appear to be drug type-, dose-, and culture time-dependent. However, further studies are needed to conclude a consistent outcome for the effects of bisphosphonate on differentiation potential of AFMSCs.
Alendronate ; Alkaline Phosphatase ; Amniocentesis ; Amniotic Fluid ; Bone Diseases, Metabolic ; Bone Resorption ; Diphosphonates ; Female ; Humans ; Imidazoles ; Karyotyping ; Mesenchymal Stromal Cells ; Osteoblasts ; Osteoclasts ; Pregnancy ; Pregnancy Trimester, Second ; Pregnant Women

OBJECTIVE: The incidence of subarachnoid hemorrhage (SAH) worldwide varies considerably. In spite of many reports about the incidence of SAH, there has been no report about the incidence of SAH on the basis of the Korean population. The purpose of this hospital-based study was to assess the actual incidence rates of aneurysmal SAH in Gwangju city and Jeollanamdo province. METHODS: All cases of SAH confirmed by computerized tomography (CT) between January 2007 and December 2007 were selected for analysis. For the data collection, three major training hospital and ten general hospitals working the CT in Gwangju city and four major general hospitals in Jeollanamdo province participate in this study. RESULTS: According to the official census of Korea, the population was 1,413,444 in Gwangju city and 1,929,836 in Jeollanamdo province in 2007. There were 163 patients in Gwangju city and 266 patients in Jeollanamdo province confirmed SAH by CT in 2007. The crude and the age- and sex-adjusted annual incidence rates per 100,000 population for all ages in Gwangju city were 11.5 and 12.4 for aneurysmal SAH and in Jeollanamdo province were 13.8 and 10.8. The incidence was higher in women and increased with age. The gender distribution varied with age. At young ages, the incidence was higher in men while after the age of 40 years, the incidence was higher in women. CONCLUSION: In the present study, the age- and sex-adjusted annual incidence rates is 11.8 in Gwangju city and Jeollanamdo province. The incidence was higher in women and increased with age.
Aneurysm ; Censuses ; Data Collection ; Female ; Hospitals, General ; Humans ; Incidence ; Korea ; Male ; Risk Factors ; Subarachnoid Hemorrhage

BACKGROUND: Leptin has been found to be involved in the regulation of hematopoiesis processes and angiogenesis. Therefore, we investigated the effect of leptin in the proliferation and angiogenesis of peripheral blood (PB)-derived endothelial progenitor cells (EPCs). METHODS: Mononuclear cells were isolated from PB of healthy male volunteers and were cultured in endothelial cell growth medium-2 (EGM-2). After 6 days of culture, cells were treated with 50 ng/mL vascular endothelial growth factor (VEGF) and/or with various concentrations of leptin (10 ng/mL, 100 ng/mL, 1microgram/mL, and 10 microgram/mL) and were further cultured for one week. Proliferation of EPCs was examined by an assay measuring the uptake of 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbo cyanine-labeled acetylated LDL (Dil-ac-LDL) and tubule formation on a matrigel. The control group of cells was not treated with VEGF and/or leptin. RESULTS: The number of Dil-ac-LDL labeled-EPCs, tubule formation on matrigel and the number of cells present along tubules were significantly increased in the leptin-treated groups of cells as compared to the control group or VEGF treated group of cells (P<.05). The effect was synergistically increased in the group of cells co-treated with leptin and VEGF. The number of EPCs was increased in a leptin dose-dependent manner that was maximal at a concentration of 1microgram/mL leptin. CONCLUSION: This study shows that leptin increased in vitro proliferation and angiogenesis of EPCs derived from peripheral blood.
Endothelial Cells ; Hematopoiesis ; Humans ; Leptin* ; Male ; Stem Cells* ; Vascular Endothelial Growth Factor A ; Volunteers