Microarray Analysis of Gene Expression During Differentiation of Human Mesenchymal Stem Cells Treated with Vitamin E in vitro into Osteoblasts

OBJECTIVE: Supplementation with vitamin E is able to protect bone against free radical-induced elevation of bone-resorbing cytokines. We examined gene expression by microarray analysis during the differentiation of human mesenchymal stem cells treated with vitamin E into osteoblasts in vitro. METHODS: Human bone marrow stem cells were cultured in osteogenic differentiation medium and vitamin E was added. A colorimetric immunoassay for the quantification of cell proliferation was used to measure osteoblast differentiation. Gene expression was analyzed using a microarray technique. We also used a real time reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: It was found that vitamin E enhanced cell proliferation when compared to cells cultured in media without vitamin E. We focused on 68 genes which are related to osteogenesis and osteoclastogenesis. Alkaline phosphatase, transforming growth factor-beta 1, fibroblast growth factor receptor 1, matrix metalloproteinase 2, muscle segment homeobox 2, bone morphogenetic protein 1, biglycan, vascular endothelial growth factor B, dentin sialophosphoprotein, cartilage oligomeric matrix protein, runt-related transcription factor 2, fibroblast growth factor receptor 3, and SMAD2 were upregulated > 2-fold compared to the control. Conversely, osteopetrosis-associated transmembrane protein 1, microphthalmia-associated transcription factor, and epidermal growth factor receptor were downregulated > 2-fold compared to the control. Vitamin E produced a 1.5-fold increase in the expression of runt-related transcription factor 2 and transforming growth factor-beta 1 as determined by real time RT-PCR. CONCLUSION: Vitamin E had a positive effect on the gene expressions regarding osteogenic differentiation of mesenchymal stem cells.

Barium Stimulates the Expression of Osteogenic Genes in Human Mesenchymal Stem Cells into Osteoblasts in vitro / 대한폐경학회지

OBJECTIVES: The purpose of this study was to determine the effect of barium on gene expression in differentiation of human mesenchymal stem cells into osteoblasts in vitro. METHODS: Human bone marrow stem cells were cultured for 0~14 days in osteogenic differentiation medium with strontium chloride (SrCl2) and barium chloride (BaCl2). Alkaline phosphatase (ALP) activity staining was the method selected for measuring osteoblast differentiation. Total ribonucleic acid (RNA) was extracted after 1, 3, 7, and 14 days, and analysis of runt-related transcription factor 2/core-binding factor alpha 1 (Runx2/Cbfa1), bone morphogenetic protein-2 (BMP-2), and bone sialoprotein (BSP) gene expression was performed by real-time reverse transcriptase (RT)-polymerase chain reaction (PCR). RESULTS: Barium and strontium had a superior enhancing effect on cell proliferation when compared to cells cultured in media without strontium or barium. BaCl2 produced a 2-fold increase in the expression of Runx2/Cbfa1 at 14 days. SrCl2 (0.1~0.3 mM) produced a 2-fold increase in the expression of Runx2/Cbfa1 at 14 days. Barium produced a 1.5-fold increase in the expression of BMP-2 on days 1 or 3. Expression of BSP was increased 1.5~1.7- and 2-fold on days 1 and 14 by barium and strontium, respectively. CONCLUSION: Barium-like strontium is considered one of the important factors in inducing mesenchymal stem cells to differentiate into osteoblasts with further enhancement on bone formation.

Comparison of proteomes in various human plasma preparations by two-dimensional gel electrophoresis.

Serum or plasma can be utilized in a variety of studies targeted toward the discovery of disease biomarkers. In this study, the proteome profiles of plasma samples prepared using various anticoagulants (EDTA, heparin or citrate), were compared with those of serum using two-dimensional electrophoresis (2-DE). Proteins which evidenced different levels in the plasma and serum were screened and identified using ESI-Q-TOF MS/MS. The proteins which became detectable after the removal of fibrinogen from serum were identified as pigment epithelial differentiating factor (four spots), fetuin-like protein, and the hemopexin precursor. In particular, three proteins, pre-serum amyloid P component, plasma glutathione peroxidase precursor, and tetranectin, evidenced increased volume intensity only in the plasma samples prepared with EDTA.

Three-dimensional gestational sac volumetry with automatic tracing mode of the VOCAL-imaging program as the new biometric parameter in determining gestational age: Preliminary study / 대한산부인과학회지

OBJECTIVE: The purpose of this study is to verify the correlation between gestational sac volume (GSV) from automatic tracing mode the VOCAL-imaging program and gestational age and to construct the nomogram of gestational sac volume as the new biometric parameter in early pregnancy. METHODS: The cross-sectional study has been conducted in 242 uncomplicated singleton pregnancies. 47 cases were excluded due to early pregnancy failure, fetal malformations, elective abortion, age discrepancy, etc. In 195 uncomplicated singleton pregnancies from 5 to 12 weeks' menstrual age, gestational sac volume, mean sac diameter and crown-lump length were measured for the assessment of gestational age. Gestational sac volumetry was carried out with automatic method and manual method of the VOCALTM (Virtual Organ Computer- aided AnaLysis) for the comparison between two methods. The collected data were analyzed for mean, standard deviation, 90% reference interval, 5th, 50th and 95th percentiles of gestational sac volume, mean sac diameter and CRL, and the nomogram were constructed. RESULTS: Polynomial regression analysis demonstrated the statistically significant positive correlation between gestational age and gestational sac volume by automatic tracing mode (R2 0.826, p<0.001), gestational sac volume by manual tracing mode (R2 0.844, p<0.0001), mean sac diameter (R2 0.763, p<0.0001) and crown-lump length (R2 0.950, p<0.0001). The 5th, 50th and 95th percentiles of the gestational sac volume were calculated and the nomogram was tabulated. In determining gestational age, the standard deviation (SD) of gestational sac volume by automatic tracing mode is 5.6 days, the SD of gestational sac volume by manual tracing mode is 5.2 days and the SD of MSD is 6.6 days. CONCLUSION: we can conclude that three-dimensional GSV with automatic tracing mode of the VOCAL-imaging program can be used as the new biometric measurement in determining gestational age. Gestational sac volumetry with automatic tracing mode of the VOCAL-imaging program have been proven available and convenient method and it can be recommended in 5-7 weeks' of gestation, when CRL is not clearly visualized.

The efficacy of HPV DNA chip test in the atypical squamous cell of undetermined significance / 부인종양

OBJECTIVE: This study was performed to investigate the efficacy of DNA chip method for detecting and genotyping of human papillomavirus (HPV) and screening of high-grade CIN (cervical intraepithelial neoplasia) or invasive cancer in the patients with atypical squamous cells of undetermined significance (ASC-US). METHODS: This study was based on 131 cases to be revealed ASC-US by Pap smear for the cervical cancer screening from July 2004 to Octorber 2004. They were evaluated by HPV DNA chip test, Cervical colposcopy and directed biopsy, and cone biopsy. The results of type 16, 18, 31, 33, 35, 39, 45, 51, 52, 53, 54, 56, 58, 59, 66 and 68 in HPV DNA chip test were categorized as high risk HPV. We evaluate the detection rate of the high-grade CIN and invasive cancer by HPV DNA chip test. RESULTS: The incidence of high risk HPV DNA was 51.1% (67/131). Twelve of 131 (9.2%) were diagnosed as high-grade CIN or CIS on histology. The detection rate of high risk HPV DNA in high-grade CIN and invasive cancer was 83.3% (10/12). The detection rate of high risk HPV DNA was 36.0% (31/86) in normal or reactive, and 83.3% (10/12) in CIN II or above on histology. Higher the grade of biopsy, more the detection rate of high risk HPV DNA by HPV DNA chip test. CONCLUSION: The use of HPV DNA chip test in patients with ASC-US may be useful in detection of high-grade CIN or invasive cancer.

The relationship between pregnancy rate and suppression of luteinizing hormone in IVF-ET program / 대한산부인과학회지

OBJECTIVE: To investigate the effect of profound LH suppression in the late follicular phase during controlled ovarian hyperstimulation (COH) on pregnancy rate. METHODS: Data were collected by retrospective analysis. A total of 172 cycles of oocytes retrieval for IVF after COH with GnRH agonist-down regulation and hMG/FSH were included from January 1999 to July 2002. Serum LH, FSH, estradiol and progesterone concentrations were measured on the day 3 of the preceding cycle and LH concentration was measured on the day of hCG administration. Based on LH concentrations (mIU/mL) on the day of hCG administration, cycles are classfied to two groups (group 1: LH>0.7 mIU/mL, group 2: LH