The uptake of glucose for metabolism and growth is essential to most animal cells and is mediated by glucose-transporter (GLUT) proteins. The aim of this study was to determine which class of glucose transporter molecules was responsible for uptake of glucose in the mouse early embryo and at which stage the corresponding genes were expressed. In addition, co-culture system with vero cell was used to investigate the effect of the system on GLUT expression. Two-cell stage embryos were collected from the superovulated ICR female and divided into 3 groups. As a control, embryos were cultured in 0.4% BSA-T6 medium which includes glucose. For the experimental groups, embryos were cultured in either co-culture system with vero cells or glucose-free 76 medium supplemented with 0.4% BSA and pyruvate as an energy substrate. 2-cell to blastocyst stage embryos in those groups were respectively collected into microtubes (50 embryos/tube). Total RNA was extracted and RT-PCR was performed. The products were analysed after staining ethidium bromide by 2% agarose gel electrophoresis. Blastocysts were collected from each group at 120hr after hCG injection. They were fixed in 2.5% glutaraldehyde, stained with hoechst, and mounted for observation. In control, GLUT1 was expressed from 4-cell to blastocyst. GLUT2 and GLUT3 were expressed in morula and blastocyst. GLUT4 was expressed in all stages. When embryos were cultured in glucose-free medium, no significant difference was shown in the expression of GLUTI1, 2 and 3, compared to control. However GLUT4 was not expressed until morular stage. When embryos were co-cultured with vero cell, there was no significant difference in the expression of GLUT1, 2, 3 and 4 compared to control. To determine cell growth of embryos, the average cell number of blastocyst was counted. The cell number of co-culture (93.8+/-3.1, n=35) is significantly higher than that of control and glucose-free group (76.6 +/- 3.8, n=35 and 68.2+/-4.3, n=30). This study shows that the GLUT genes are expressed differently according to embryo stage. GLUTs were detectable throughout mouse preimplantation development in control and co-culture groups. However, GLUT4 was not detected from 2- to 8-cell stage but detected from morula stage in glucose-free medium, suggested that GLUT genes are expressed autocrinally in the embryo regardless of the presence of glucose as an energy substrate. In addition, co-culture system can increase the cell count of blastocyst but not improve the expression of GLUT. In conclusion, expression of GLUT is dependent on embryo stage in preimplantation embryo development.
Animals ; Blastocyst ; Cell Count ; Coculture Techniques ; Electrophoresis, Agar Gel ; Embryonic Development ; Embryonic Structures* ; Ethidium ; Female ; Glucose Transport Proteins, Facilitative* ; Glucose* ; Glutaral ; Humans ; Metabolism ; Mice* ; Morula ; Pregnancy ; Pyruvic Acid ; RNA ; Vero Cells

OBJECTIVES: We performed anterior spinal fusion and instrumentation in treatment of spinal tuberculosis. The clinical results of this operation and metal-related complications were evaluated to determine the rationale of anterior instrumentation in active tuberculous lesion. METHODS: From July 1989 to February 1993, we treated twenty-one patients with spinal tuberculosis by radical resection of the tuberculous lesion and bone grafting, followed by anterior instrumentation using Zielke rod system. The changes in spinal deformity were measured from lateral spinal radiographs obtained preoperatively and postoperatively at 3 months, 6 months, 1 year, 2 years and final follow-up. The recurrence of infection and possible complications were also observed clinically and radiologically. RESULTS: The mean kyphotic angle was decreased preoperatively from 21 degrees to 16 degrees at final follow-up in patients with thoracolumbar tuberculous lesions. The mean deformity angle was corrected 7 degrees in thoracolumbar tuberculosis and 12 degrees in lumbar tuberculosis compared with the preoperative deformity angle. There was not any persistence or recurrence of infection possibly related to the instrumentation. All patients were allowed early ambulation with the aid of a light brace. CONCLUSION: The clinical and radiological results suggested that the anterior instrumentation seemed to be one of the rational approaches for providing immediate stability in treating severe spinal tuberculosis without any significant risk of persistence or recurrence of infection.
Bone Transplantation ; Braces ; Congenital Abnormalities ; Early Ambulation ; Follow-Up Studies ; Humans ; Recurrence ; Spinal Fusion ; Tuberculosis ; Tuberculosis, Spinal*

SUMMARY: The present study was carried out to evaluate whether the coculture system of human embryos with Vero cells can improve the quality of embryo or overcome the repetitive implantation failures in order to obtain pregnancy. From January to December 1996, a total 202 cases which patients with the problems of repetitive implantation failures (group I) or those with the poor embryonic quality in their previous cycles (group II) was analysed. The quality of cocultured embryo, pregnancy, on-going and implantation rates between coculture and control groups were compared. Of 93 cases in group I, coculture was performed in 34 cases and conventional IVF for the rest. Of 109 cases in group II, 36 for coculture and 73 for conventional IVF. In group I, pregnancy, on-going and implantation rates in coculture group (14/34 (41.2%), 9/34 (26.5%), 16/81 (19.8%), respectively) were higher than those of control (11/59 (18.6%), 8/59 (13.6%), 12/152 (7.9%), respectively). There is significance in the pregnancy and implantation rates (p=0.028 and p=0.015). In group II, pregnancy, on-going and implantation rates in coculture group (8/36 (22.2%), 5/36 (13.9%), 8/87 (9.2%), respectively) were higher than those of control (5/73 (6.8%), 3/73 (4.1%), 3/158 (1.9%), respectively). Like the result of group 1, there is significance in the pregnancy and implantation rates (p=0.028 and p=0.022). Coculture system with Vero cells works well in the groups of the two indications. Although the case of 3 day-coculture was small as 15 cases in group II, 3 day-coculture improved pregnancy rate (4/15 (26.7%)). Therefore, 3 day-coculture with assisted hatching is recommended to the patients with poor embryonic quality. In conclusion, coculture system with Vero cells can be suggested as an effective method which improves pregnancy rate in those who have repetitive implantation failures or whose embryonic quality was poor in their previous cycles.
Coculture Techniques* ; Embryonic Structures ; Humans ; Humans* ; Pregnancy ; Pregnancy Rate ; Prognosis* ; Vero Cells

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.
Embryo Transfer* ; Embryonic Structures* ; Freezing ; Humans ; Infertility ; Insemination ; Male ; Pregnancy Rate* ; Pregnancy* ; Sperm Injections, Intracytoplasmic ; Spermatozoa ; Survival Rate ; Zygote

PURPOSE: To evaluate the biologic changes of xenograft (Lubboc, Transphyto S. A., Clermont-Ferrand, France) applied in the surgical treatment of calcaneal fractures. MATERIALS AND METHODS: Xenograft were used in twenty-five calcaneal fractures, as bone substitutes for mechanical support of the articular surface from March 1994 to December 1996. Twenty-four cases had obtained clinical and radiological bony union without complications. We performed biopsy at the xenograft area in eight cases when the hardware was removed. And then we took a magnetic resonance imaging of the calcaneus after the biopsy in three cases. RESULTS: The pathologic findings of the xenografts showed dead bony trabeculae with inflammed fibrous tissue. There was no osteoblastic activity but some osteoclastic activity was seen in the graft-host junctional area. MRI findings showed that there was well-circumscribed, low-signal intensity in the xenograft area contrast to the normal bony signal intensity. Its lesion was equal to the sclerotic finding in the calcaneal radiograph. CONCLUSIONS: Xenograft (LUBBOC'), which is used in the mechanical support of the articular surface, apparently lacks osteogenic capacity.
Biopsy ; Bone Substitutes ; Calcaneus ; Heterografts* ; Magnetic Resonance Imaging ; Osteoblasts ; Osteoclasts

PURPOSE: The results of the bronchial provocation test were different by the method of inhalation, but there was no data in children. We performed the provocation test by 2 different methods such as two-minute tidal breathing versus five vital capacity breathing and compared the datas. METHODS: Methacholine inhalation test was performed for the In 17 patients with suspected bronchial hyperresponsiveness in Pocheon university Bundang Cha hospital, under identical conditions, except for the method of inhalation. All the patients were performed two-minute tidal breathing (2 min-TB) in one test and five vital capacity breathing (5VCB) with interval 1 to 7 days. Baseline and postsaline FEV1 were also measured to compare effects by two inhalation methods. Methacholine PC20 was calculated by linear interpolation between the last two data parts on the dose-response curve, and the cut off value of bronchial asthma was 8 mg/mL. RESULTS: The rate of positive brochial provaction test in 5VCB was 88.24% which was not significantly lower than 70.59% in 2 min-TB. In 12 patients with positive bronchial provocation test in both inhalation methods, the mean PC20 in 5VCB (5VC-PC20) was 4.40 mg/mL, being significantly higher than 1.02 mg/mL in 2 min-TB (2 min-PC20) (P< 0.05). There was a significant positive correlation between 5VC-PC20 and 2 min-PC20 (R=0.76, P< 0.001). CONCLUSION: We conclude from this study that the results of bronchial provocation test in children are different according to methods of inhalation in using the same nebulizer. Therefore, when we analyze the result of bronchial provocation test, we need attention to factors which are nebulizer output and method of inhalation to influence the response to the test.
Asthma ; Bronchial Provocation Tests* ; Child* ; Humans ; Inhalation ; Methacholine Chloride ; Nebulizers and Vaporizers ; Respiration* ; Vital Capacity*

alpha-Viniferin (AVF), a trimer of resveratrol, is known to have an anti-inflammatory effect via inhibition of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS). It has been reported that up-regulated COX-2 and iNOS are expressed in colon cancer tissues of humans and rodents as well as pre-neoplastic aberrant crypt foci (ACF) of rodents. In this study, chemopreventive effects of AVF were assessed in Caco-2 cells as well as azoxymethane (AOM)-induced colorectal tumorigenesis in mice. Anti-tumor effect of AVF with regards to apoptotic induction was assessed by TUNEL and caspase-3 expression in human colon cancer Caco-2 cells. For development of ACF, AOM was administered with to mice intraperitoneally at a dose of 10 mg/kg once a week for 3 weeks. To induce colitis-related colon cancer, mice were administered a single dose of AOM (10 mg/kg) and 2% dextran sodium sulfate in drinking water. Mice treated with 0.05 and/or 0.1 mg of AVF by gavage showed significantly reduced development of ACF and colorectal tumors. Immunofluorescence detection in Caco-2 cells showed reduced COX-2 and iNOS expression, whereas cleavage of caspase-3 and apoptotic cell numbers increased upon AVF treatment. Immunostaining showed reduced expression levels of COX-2 and iNOS expression along with increased cleaved caspase-3 expression increased upon AVF treatment. These results suggest that AVF has chemopreventive effects on colorectal cancer via anti-inflammatory potential and pro-apoptotic activity.
Aberrant Crypt Foci ; Animals ; Azoxymethane ; Caco-2 Cells* ; Carcinogenesis ; Caspase 3 ; Cell Count ; Chemoprevention ; Colonic Neoplasms ; Colorectal Neoplasms* ; Cyclooxygenase 2 ; Dextrans ; Drinking Water ; Fluorescent Antibody Technique ; Humans ; In Situ Nick-End Labeling ; Mice* ; Nitric Oxide Synthase Type II ; Rodentia ; Sodium

BACKGROUND: This study was conducted to analyze the current problems in completing death certificates and to identify the correct method for completing death certificates. METHODS: We reviewed 262 death certificates in three hospitals from March 1 to April 30, 2000, and 119 death certificates in one hospital from March 1 to 31, 2000. We identified major and minor errors and analyzed and compared them retrospectively. RESULTS: A total of 381 death certificates were reviewed: 59 in Seoul National University Hospital, 101 in Ewha Woman's University Hospital, and 102 in Gachon Medical College Hospital, which has no education program for completing death certificates in postgraduate training, and 119 in Samsung Medical Center which has an education program for completing death certificates. 358 certificates(94.0%) had at least one error. There were only 23 death certificates(6.0%) without an error. In 182 cases(47.8%), there was one major error. In 321 death certificates(84.3%), there were more than two errors. A comparison of Samsung Medical Center with the other hospitals showed that the number of total errors was statistically different(p=0.001). CONCLUSION: There were few death certificates without an error in this study. In a hospital which has postgraduate training in completing death certificates, there are fewer errors than in other hospitals which have no training course. Emergency physicians actually certify many deaths, so they must know the correct method of completing death certificates for statistics on morbidity and mortality.
Death Certificates* ; Education ; Emergencies ; Mortality ; Retrospective Studies ; Seoul

OBJECTIVE: The aim of this study was to analyze the effect of supplementing vitrification and warming solutions with two types of antifreeze proteins (AFPs) and the combination thereof on the follicular integrity of vitrified-warmed mouse ovaries. METHODS: Ovaries (n=154) were obtained from 5-week-old BDF1 female mice (n=77) and vitrified using ethylene glycol and dimethyl sulfoxide with the supplementation of 10 mg/mL of Flavobacterium frigoris ice-binding protein (FfIBP), 10 mg/mL of type III AFP, or the combination thereof. Ovarian sections were examined by light microscopy after hematoxylin and eosin staining, and follicular intactness was assessed as a whole and according to the type of follicle. Apoptosis within the follicles as a whole was detected by a terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick-end labeling assay. RESULTS: The proportion of overall intact follicles was significantly higher in the type III AFP-supplemented group (60.5%) and the combination group (62.9%) than in the non-supplemented controls (43.8%, p<0.05 for each). The proportion of intact primordial follicles was significantly higher in the FfIBP-supplemented (90.0%), type III AFP-supplemented (92.3%), and combination (89.7%) groups than in the non-supplemented control group (46.2%, p<0.05 for each). The proportions of non-apoptotic follicles were similar across the four groups. CONCLUSION: Supplementation of the vitrification and warming solutions with FfIBP, type III AFP, or the combination thereof was equally beneficial for the preservation of primordial follicles in vitrified mouse ovaries.
Animals ; Antifreeze Proteins* ; Apoptosis ; Deoxyuridine ; Dimethyl Sulfoxide ; DNA Nucleotidylexotransferase ; Eosine Yellowish-(YS) ; Ethylene Glycol ; Female ; Fertility Preservation ; Flavobacterium ; Hematoxylin ; Humans ; Mice* ; Microscopy ; Ovary* ; Vitrification

BACKGROUND: The abnormal barrier function in atopic dermatitis(AD) is caused by a reduction in the amounts of ceramides in the intercellular lipids in the stratum corneum(SC). Replenishing the SC via the topical application of ceramides and pseudoceramides leads to effective recovery of the barrier function of skin. OBJECTIVES: An open clinical crossover evaluation was conducted to investigate the effects in AD of a multi-lamellar emulsion(MLE) that contained pseudoceramide(PC-9s). METHODS: The study group included 30 AD patiendaverage age: 4.4 yr, range: 1-8 yr), who applied MLE or a commercial moisturizing cream(CMC, 5% urea) alternately for four weeks each. We divided the subjects into two subgroups and started with different treatments in each subgroup. Treatment efficacy was evaluated using the average subjective satisfaction scores for each symptom and the global clinical response. In addition, the SCORAD(Scoring AD) index was adopted to evaluate the severity of AD as objectively as possible. The patients were evaluated using this index every other week. RESULTS: Although the SCORAD improved in both subgroups, the patients had better results (p<0.05) when applying MLE(31-35% decrease) than CMC(13% increase to 14% decrease). The subjective satisfaction scores of the symptoms and signs of patients, including itching, erythema, and dry skin, were higher in the MLE group than in the CMC group, and the global response to treatment was also better in the MLE group. During the follow-up period, AD improved in all patients. MLE was more effective than CMC in our patients. CONCLUSIONS: The topical application of a multi-lamellar emulsion containing pseudoceramide is an effective regimen for improving symptoms of AD.
Ceramides ; Cross-Over Studies* ; Dermatitis, Atopic* ; Erythema ; Follow-Up Studies ; Humans ; Pruritus ; Skin ; Treatment Outcome