OBJECTIVE: Human embryonic stem (ES) cells have a great potential in regenerative medicine and tissue engineering. The human ES cells could be differentiated into specific cell types by treatments of growth factors and alterations of gene expressions. However, the efficacy of guided differentiation and isolation of specific cells are still low. In this study, we characterized isolated cells from differentiated human ES cells by magnetic activated cell sorting (MACS) system using specific antibodies to cell surface markers. METHODS: The undifferentiated hES cells (Miz-hESC4) were sub-cultured by mechanical isolation of colonies and embryoid bodies were spontaneously differentiated with DMEM containing 10% FBS for 2 weeks. The differentiated cells were isolated to positive and negative cells with MACS system using CD34, human epithelial antigen (HEA) and human fibroblast (HFB) antibodies, respectively. Observation of morphological changes and analysis of marker genes expression were performed during further culture of MACS isolated cells for 4 weeks. RESULTS: Morphology of the CD34 positive cells was firstly round, and then it was changed to small polygonal shape after further culture. The HEA positive cells showed large polygonal, and the HFB positive spindle shape. In RT-PCR analysis of marker genes, the CD34 and HFB positive cells expressed endodermal and mesodermal genes, and HEA positive cells expressed ectodermal genes such as NESTIN and NF68KD. The marker genes expression pattern of CD34 positive cells changed during the extension of culture time. CONCLUSION: Our results showed the possibility of successful isolation of specific cells by MACS system from undirected differentiated human ES cells. Thus, MACS system and marker antibodies for specific cell types might be useful for guided differentiation and isolation of specific cells from human ES cells.
Antibodies ; Ectoderm ; Embryoid Bodies ; Endoderm ; Fibroblasts ; Gene Expression ; Humans* ; Intercellular Signaling Peptides and Proteins ; Mesoderm ; Nestin ; Regenerative Medicine ; Tissue Engineering

Background: Stress as a cause of mental health problems is known to be more prevalent in women than in men and has a negative effect on several aspects of physical health, such as the composition of blood and saliva. This study investigated the relationship of perceived stress with blood cell counts, saliva flow rate, and saliva factors. Methods: We recruited women in their 20s with a high prevalence of stress. Stress was evaluated using the Korean version of the perceived stress scale. Blood tests included white blood cell, hemoglobin, and platelet. We then examined the saliva flow rate and cariogenic bacteria level, acidity, occult blood, buffer capacity, leukocyte level, protein level, and ammonia level using rinse water with the SILL-HaⓇ saliva test system. Results: In a total of 70 participants, the average age was 21.64 years old, the average perceived stress score was 16.96±4.32, and high levels of stress were reported by 80% of the participants (n=56). The high-stress group had lower hemoglobin levels. In addition, the high-stress group showed a lower saliva flow rate than the low-stress group, and there was a difference in the salivary acidity and buffer capacity. The total perceived stress score showed a positive correlation with acidity and negative correlation with buffer capacity and the hemoglobin level. Conclusion This study found that stress in female college students might affect the composition of blood and saliva. High levels of stress were positively correlated with the hemoglobin level, saliva flow rate, and acidity and negatively correlated with the buffer capacity.

Purpose: This study was conducted to determine the factors affecting a hypothetical model of testing for general hospital nurses’ job embeddedness. Methods: Data were collected from August 20 to September 19, 2021, by a self-administered questionnaire answered by 428 general hospital nurses. The data were analyzed by SPSS and AMOS. Results: Nine of the hypothetical model’s 12 hypotheses were supported by the data collected from all participants. The test results indicate that ego resilience, subjective career success, and recovery experience from job stress directly affect participants’ job embeddedness. Nurses’ work environments were reported to affect ego resilience and subjective career success, while at the same time ego resilience and subjective career success affected the participants’ job embeddedness. Work-life balance was found to affect ego resilience and ego resilience affected subjective career success, and at the same time subjective career success directly affected participants’ recovery experiences from job stress and job embeddedness. Of these variables, subjective career success had the strongest direct effect on participants’ job embeddedness. Work-life balance affected the participants’ recovery experiences from job stress, and their recovery experiences from job stress were found to directly affect job embeddedness. Conclusion These results suggest that different management strategies to enhance hospital nurses’ job embeddedness should address nurses’ ego resilience, subjective career success, and recovery experiences from job stress.

No abstract available.

One author's affiliation is misspelled in original article.

To identify strategies to prevent sexual problems in teenage girls, respondents in this study answered two open-ended questions: "What are strategies for teenage school girls to prevent unwanted coitus?" and "What are strategies for teenage girls to prevent pregnancy?" The respondents were 12,733 girls from an accessible population of 19,000, a multi-stage cluster sample from a population of 1,988,902 girls attending 4,684 schools in the seven largest cities and nine provinces in Korea. Data were collected by mail between October 2 and October 28, 2000. The response rate was 68.9%. The total number of responses for the first question was 10,345, and for the second, 9,624. Data were analyzed by content analysis. The results of this study are: 1. According to priority, frequent strategies to prevent unwanted coitus were, self assertiveness (35.7%), heterosexual interaction training (24.6%), sex education (21.2%), and innovations in the system of social culture (4.7%). The order of priority was the same whether the respondents had experienced coitus or not. 2. According to priority, frequent strategies to prevent pregnancy were, heterosexual interaction training (27.4%), sex education (26.2%), contraceptive use and induced abortion (21.4%), and innovations in the system of social culture (3.2%). The first priority for the respondents who had not experienced coitus was heterosexual interaction training (27.7%) but contraceptive use (35.5%) was the first priority for the group who had experienced coitus. In sex education, a focus on contraceptive use for teenage girls who have experienced coitus and on heterosexual interaction training for those who have not, would strengthen preventive strategies for these two sexual problems. Assertiveness training as part of sex education would further strengthen prevention strategies.
Abortion, Induced ; Assertiveness ; Coitus ; Surveys and Questionnaires ; Female* ; Heterosexuality ; Humans ; Korea* ; Postal Service ; Pregnancy ; Sex Education

OBJECTIVES: The purpose of this study was to determine whether or not the shed endometrial tissues in menstrual fluid (MF) have adhesive potentials, using human amniotic membrane (AM). METHODS: The MF from 20 patients with regular menstruation was collected with Wallace catheter by aspiration from the uterine cavity on the second or third day of the menstrual period. The AM was obtained from the placenta of term delivery without any complication. The MF was washed and diluted fivefold with Hams F-10 medium supplemented with 10% fetal bovine serum. The cell suspension was placed on either epithelial layer (EP) or extracellular matrix layer (ECM) of the AM. After 5 days of culture, the adhesion sites were observed under a stereomicroscope. For histological observation, each cultured AM was prepared for the serial paraffin section. RESULTS: The adhesion sites of endometrial tissues in MF were found both ECM (20/20) and EP (11/20) of the AM. The size of adhesion sites in each AM were highly variable from microscopic to macroscopic size. CONCLUSION: We found that the shed endometrial tissues in MF have adhesive potential to epithelial layer in addition to extracellular matrix layer of amniotic membrane. This adhesive potential may be related to pathogenesis of endometriosis. We suggest that this culture system can be useful as an in-vitro model for endometriosis.
Adhesives* ; Amnion* ; Catheters ; Endometriosis* ; Extracellular Matrix ; Female ; Humans ; Menstruation ; Paraffin ; Placenta

No abstract available.
Epidermolysis Bullosa ; Muscular Dystrophy, Duchenne ; Ornithine Carbamoyltransferase ; Polymerase Chain Reaction* ; Preimplantation Diagnosis*

OBJECTIVE: To evaluate the effects of recombinant FSH (rFSH) and urinary FSH (uFSH) on the gene expressions of human endometrial stromal cells in vitro. METHODS: Endometrial tissue was obtained from a pre-menopausal women undergoing hysterectomy. Primary endometrial stromal cells were isolated and in vitro cultured with FBS-free DMEM/F-12 containing 0, 10, 100, and 1,000 mIU/ml of rFSH and uFSH for 48 hours, respectively. Total RNA was extracted from the cultured cells and subjected to real time RT-PCR for the quantitative analysis of progesterone receptor (PR), estrogen receptor alpha/beta (ER-alpha/beta), cyclooxygenase 2 (Cox-2), leukemia inhibitory factor (LIF), homeobox A10-1 and -2 (HoxA10-1/-2). RESULTS: Both hormone treatments slightly increased (< 3 folds) the expressions of PR, ER-beta and HoxA10-1/-2 gene. However, ER-alpha expression was increased up to five folds by treatments of both FSH for 48 hours. The LIF expression by the 10 mIU/ml of uFSH for 12 hours was significantly higher than that of rFSH (p<0.01). After 24 hours treatment of two kinds of hormones, the expression patterns of LIF were similar. The 100 and 1,000 mIU/ml of rFSH induced significantly higher amount of Cox-2 expression than those of uFSH, respectively (p<0.05). CONCLUSION: This study represents no adversely effect of exogeneous gonadotropins, rFSH and uFSH, on the expression of implantation related genes. We suggest that rFSH is applicable for the assisted reproductive technology without any concern on the endometrial receptivity.
Cells, Cultured ; Cyclooxygenase 2 ; Estrogens ; Female ; Gene Expression* ; Genes, Homeobox ; Gonadotropins ; Humans* ; Hysterectomy ; Leukemia Inhibitory Factor ; Receptors, Progesterone ; Reproductive Techniques, Assisted ; RNA ; Stromal Cells*

OBJECTIVE: To investigate the association between endometriosis and polymorphisms of N-acetyl transferase 2 (NAT2), glutathione S-transferase M1 (GSTM1), and cytochrome P450 (CYP) 1A1 genes in Korean infertile patients. MATERIALS AND METHODS: A total of 303 infertile patients who had undertaken diagnostic laparoscopy during January, 2001 through December, 2003 at Samsung Cheil Hospital enrolled in this study. The patients were grouped according to laparoscopic findings: minimal to mild endometriosis (group I: n=147), moderate to severe endometriosis (group II: n=57), normal pelvic cavity (n=99). Peripheral blood was obtained and genomic DNA was extracted. The genotypes of each genes were analyzed using polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP). For NAT2, RFLP was used to detect the wild type (wt) and mutant (mt) alleles, enabling classification into slow (mt/mt) or fast (wt/wt or wt/mt) acetylation genotypes. For GSTM1, PCR was used to distinguish active (+/- or +/+) from null (-/-) genotypes. For CYP1A1, MspI digestion was used to detect the wild type (A1A1), heterozygote (A1A2) or mutant (A2A2) genotypes. RESULTS: The genotype frequencies of NAT2 slow acetylator was 12.8%, 10.9%, 12.8% in group I, group II and control, respectively. The genotype frequencies of GSTM1 null mutation was 55.3%, 41.8%, 53.2% in group I, group II and control, respectively. The genotype frequencies of CYP1A1 MspI polymorphism was 16.3%, 9.1%, 18.1% in group I, group II and control, respectively. No significant difference was observed between endometriosis and normal controls in the genotype frequencies of the NAT2, GSTM1, CYP1A1 MspI polymorphism. CONCLUSION: The NAT2, GSTM1, CYP1A1 gene polymorphism may not be associated with the susceptibility of endometriosis in Korean women.
Acetylation ; Alleles ; Classification ; Cytochrome P-450 CYP1A1 ; Cytochrome P-450 Enzyme System* ; Cytochromes* ; Digestion ; DNA ; Endometriosis* ; Female ; Genotype ; Glutathione Transferase* ; Glutathione* ; Heterozygote ; Humans ; Laparoscopy ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Transferases*