Various somatic cell nuclear transfer (SCNT) techniques for mammalian species have been developed to adjust species-specific procedures to oocyte-associated differences among species. Species-specific SCNT protocols may result in different expression levels of developmentally important genes that may affect embryonic development and pregnancy. In the present study, porcine oocytes were treated with demecolcine that facilitated enucleation with protruding genetic material. Enucleation and donor cell injection were performed either simultaneously with a single pipette (simplified one-step SCNT; SONT) or separately with different pipettes (conventional two-step SCNT; CTNT) as the control procedure. After blastocysts from both groups were cultured in vitro, the expression levels of developmentally important genes (OCT4, NANOG, EOMES, CDX2, GLUT-1, PolyA, and HSP70) were analyzed by real-time quantitative polymerase chain reaction. Both the developmental rate according to blastocyst stage as well as the expression levels CDX2, EOMES, and HSP70 were elevated with SONT compared to CTNT. The genes with elevated expression are known to influence trophectoderm formation and heat stress-induced arrest. These results showed that our SONT technique improved the development of SCNT porcine embryos, and increased the expression of genes that are important for placental formation and stress-induced arrest.
Animals ; Biological Markers/metabolism ; Cloning, Organism ; Embryo, Mammalian/metabolism ; Female ; *Gene Expression Regulation, Developmental ; Nuclear Transfer Techniques/instrumentation/*veterinary ; Oocytes/metabolism ; Pregnancy ; Real-Time Polymerase Chain Reaction ; Swine/*embryology/*genetics

Alopecia has emerged as one of the biggest interests in modern society. Many studies have focused on the treatment of alopecia, such as transplantation of hair follicles or inhibition of the androgen pathway. Hair growth is achieved through proper proliferation of the components such as keratinocytes and dermal papilla cells (DPCs), movement, and interaction between the two cells. The present study examined the effect of the hedgehog (Hh) signaling pathway, which is an important and fundamental signal in the cell, on the morphology and the viability of human keratinocytes and DPCs. Upregulation of Hh signaling caused a morphological change and an increase in epithelium-mesenchymal transition-related gene expression but reduced the viability of keratinocytes, while the alteration of Hh signaling did not cause any change in DPCs. The results show the possibility that the regulation of Hh signaling can be applied for the treatment of alopecia.

Skin-derived precursors (SKPs) have potential to differentiate to various cell types including osteoblasts, adipocytes and neurons. SKPs are a candidate for cell-based therapy since they are easily accessible and have multipotency. Most mammalian cells are exposed to a low oxygen environment with 1 to 5% O2 concentration in vivo, while 21% O2 concentration is common in in vitro culture. The difference between in vitro and in vivo O2 concentration may affect to the behavior of cultured cells. In this report, we investigated the effect of hypoxic condition on stemness and proliferation of SKPs. The results indicated that SKPs exposed to hypoxic condition for 5 days showed no change in proliferation. In terms of mRNA expression, hypoxia maintained expression of stemness markers; whereas, oncogenes, such as Klf4 and c-Myc, were downregulated, and the expression of Nestin, related to cancer migration, was also downregulated. Thus, SKPs cultured in hypoxia may reduce the risk of cancer in SKP cell-based therapy.
Adipocytes ; Anoxia ; Cells, Cultured ; In Vitro Techniques ; Nestin ; Neurons ; Oncogenes ; Osteoblasts ; Oxygen ; RNA, Messenger

Human malignant melanoma is an aggressive skin cancer which has been rising at a greater rate than any other cancers. Although various new therapeutic methods have been developed in previous studies, this disease has properties of high proliferation and metastasis rate which remain obstacles that have lead to a poor prognosis in patients. It has been reported that a specific Lactobacillus extract has anti-cancer and –metastasis effect in vitro and in vivo. However, previous research has not specified precisely what effect the Lactobacillus rhamnosus GG (LGG) extract has had on human malignant melanomas. In this study, we showed that the LGG extract has anti-cancer and –metastasis effects on the human malignant melanoma cell lines, A375P and A375SM. At first, it was found that, while the LGG extract affects human neonatal dermal fibroblasts slightly, it induced the dose-dependent anti-cancer effect on A375P and A375SM by a WST-1 proliferation assay. As a result of a real-time PCR analysis, the expression patterns of several genes related to cell cycle, proliferation, and apoptosis were modulating in a manner that inhibited the growth of both malignant melanoma cell lines after the treatment of the LGG extract. Furthermore, genes related to the epithelialmesenchymal transition were down-regulated, and migration rates were also decreased significantly by the LGG extract. Our study showed that the LGG extract could be used as a potential therapeutic source.
Apoptosis ; Cell Cycle ; Cell Line ; Epithelial-Mesenchymal Transition ; Fibroblasts ; Humans* ; In Vitro Techniques ; Lactobacillus rhamnosus* ; Lactobacillus* ; Melanoma* ; Neoplasm Metastasis ; Prognosis ; Real-Time Polymerase Chain Reaction ; Skin Neoplasms

Skin-derived precursor cells (SKPs) are multipotent, sphere-forming and embryonic neural crest-related precursor cells that can be isolated from dermis. It is known that the properties of porcine SKPs can be enhanced by leukemia inhibitory factor (LIF) which is an essential factor for the generation of embryonic stem cells in mice. In our present study, to enhance or maintain the properties of murine SKPs, LIF was added to the culture medium. SKPs were treated with 1,000 IU LIF for 72 hours after passage 3. Quantitative real time RT-PCR was then performed to quantify the expression of the pluripotent stem cell specific genes Oct4, Nanog, Klf4 and c-Myc, and the neural crest specific genes Snai2 and Ngfr. The results show that the expression of Oct4 is increased in murine SKPs by LIF treatment whereas the level of Ngfr is decreased under these conditions. Interestingly, LIF treatment reduced Nanog expression which is also important for cell proliferation in adult stem cells and for osteogenic induction in mesenchymal stem cells. These findings implicate LIF in the maintenance of stemness in SKPs through the suppression of lineage differentiation and in part through the control of cell proliferation.
Adult Stem Cells ; Animals ; Cell Proliferation ; Dermis ; Durapatite ; Embryonic Stem Cells ; Leukemia ; Leukemia Inhibitory Factor ; Mesenchymal Stromal Cells ; Mice ; Neural Crest ; Pluripotent Stem Cells

OBJECTIVES: This study investigated the prescribing patterns of atypical antipsychotics for the various psychiatric disorders in the psychiatric ward of a University hospital. METHODS: We reviewed the medical records of patients who were discharged from an open psychiatric ward from May, 2003 through April, 2014. The association between psychiatric disorders and prescription pattern of atypical antipsychotics was analyzed. RESULTS: The study included 3091 patients' prescription of psychiatric medication. 60% of prescription included antipsychotics; quetiapine was the most frequently prescribed antipsychotics, but the average dosage was the lowest among all the atypical antipsychotics. According to the diagnoses, prescription rates and dosage of antipsychotics were different. Prescription rates of antipsychotics were the lowest in patients with anxiety disorders, and the mean dosage were the lowest in those with delirium, dementia, and amnestic and other cognitive disorders. CONCLUSION: This observational study shows prescription patterns of atypical antipsychotics for the treatment of psychiatric disorders in a University hospital; atypical antipsychotics were widely used for the treatment of the various disorders, and there were differences in prescription patterns for each disorders. The results of this study may be used to identify the proper atypical antipsychotics effective on certain psychiatric disorders and to propose expanding the indications of each atypical antipsychotics in the future.
Antipsychotic Agents* ; Anxiety Disorders ; Delirium ; Dementia ; Diagnosis ; Humans ; Medical Records ; Observational Study* ; Prescriptions* ; Quetiapine Fumarate

PURPOSE: This study aimed to determine health problems experienced by workers in special employment relationships (WSER) and identify coping strategies used when such problems occur.METHODS: This qualitative study used the focus group interview method. Thirteen study participants included five home-visit tutors, five insurance salespersons, and three credit card recruiters. The interviews were conducted from November 2018 through January 2019, with each occupational group interview lasting about 2 hours. Analysis based on phenomenological research was independently performed by two researchers.RESULTS: Most participants had common health problems involving vocal cord symptoms, and stress related to emotional labor and traffic accidents. The unique health problems included cystitis, musculoskeletal, and digestive symptoms in home-visit tutors; reduced vision and hearing in insurance salespersons; and mental distress in credit card recruiters. There was no protection system for their health coverage, and the company emphasized their self-employed status to avoid taking responsibility for them. Twelve participants did not purchase occupational accident insurance owing to both not having adequate information and economic burden concerning premium status.CONCLUSION: WSER experienced both physical and mental health problems. These problems were caused by their unstable employment status, and the social security system for their coverage being non-functioning.
Accidents, Occupational ; Accidents, Traffic ; Cystitis ; Employment ; Focus Groups ; Hearing ; Humans ; Insurance ; Mental Health ; Methods ; Occupational Groups ; Occupational Health ; Qualitative Research ; Social Security ; Vision, Low ; Vocal Cords

PURPOSE: This study aimed to determine health problems experienced by workers in special employment relationships (WSER) and identify coping strategies used when such problems occur. METHODS: This qualitative study used the focus group interview method. Thirteen study participants included five home-visit tutors, five insurance salespersons, and three credit card recruiters. The interviews were conducted from November 2018 through January 2019, with each occupational group interview lasting about 2 hours. Analysis based on phenomenological research was independently performed by two researchers. RESULTS: Most participants had common health problems involving vocal cord symptoms, and stress related to emotional labor and traffic accidents. The unique health problems included cystitis, musculoskeletal, and digestive symptoms in home-visit tutors; reduced vision and hearing in insurance salespersons; and mental distress in credit card recruiters. There was no protection system for their health coverage, and the company emphasized their self-employed status to avoid taking responsibility for them. Twelve participants did not purchase occupational accident insurance owing to both not having adequate information and economic burden concerning premium status. CONCLUSION WSER experienced both physical and mental health problems. These problems were caused by their unstable employment status, and the social security system for their coverage being non-functioning.

This study was performed to evaluate the effectiveness of the 2-step needle insertion technique to reduce pain during local anesthesia in children. Local anesthesia was performed bilaterally using conventional technique and a 2-step technique, and to compare the degree of pain using the 2-step with that using the conventional technique during local anesthesia using a pain scale.
The pain scores were significantly different between the conventional technique and the 2-step insertion technique at 5.82 ± 2.14 and 2.57 ± 2.09, respectively (p < 0.001). The subjective degree of pain based on gender, age, and Frankl’s behavior rating scale was significantly lower in the 2-step insertion technique (p < 0.001). Subjective pain levels in the area of maxilla and mandible, anterior and posterior parts were lower in the group treated using the 2-step insertion technique (p < 0.001).
The 2-step insertion technique is a simple and effective way to reduce pain during local anesthesia not only in children with a positive attitude to dental treatment, but also in children with negative attitudes, and is thought to be more effective if combined with application of topical anesthesia or distraction.

Background: Laboratory animals remain critical to biomedical research, despite the increasing availability of alternative approaches. Indeed, scientists strive to reduce and refine and replace the use of laboratory animals, even in the face of public calls for ever-more stringent regulation for the protection and care of animals in research. This report outlines the current status and legal regulatory issues with regard to the procurement and use of animals for research in Korea. Results: The number of animals used for education and research purposes was increased nationwide, from 2.5 to 4.9 million in 2015 and 2021, respectively. When compared with figures from the UK, institutions in Korea were found to use more mammals such as mice and dogs. In our research, we identified three major issues concerning recent animal supply in Korea, particularly: (1) Purchase of dogs from unregistered animal supplier for a dog cloning project; (2) Purchase of dogs from an unclear source for veterinary education and training; (3) Illegal cat experiments using cats obtained from unauthorized routes. Conclusions Our findings support the notion that alternatives to laboratory animal research should be implemented. We conclude that improvements in the regulations and guidelines for animal suppliers, together with the recent introduction of legislation will improve animal safety and wellbeing of animals in laboratory research in Korea.