• The Korean Journal of Physiology and Pharmacology
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• v.24 no.6
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• pp.463-472
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• 2020

Direct reprogramming, also known as a trans-differentiation, is a technique to allow mature cells to be converted into other types of cells without inducing a pluripotent stage. It has been suggested as a major strategy to acquire the desired type of cells in cell-based therapies to repair damaged tissues. Studies related to switching the fate of cells through epigenetic modification have been progressing and they can bypass safety issues raised by the virus-based transfection methods. In this study, a protocol was established to directly convert fully differentiated fibroblasts into diverse mesenchymal-lineage cells, such as osteoblasts, adipocytes, chondrocytes, and ectodermal cells, including neurons, by means of DNA demethylation, immediately followed by culturing in various differentiating media. First, 24 h exposure of 5-azacytidine (5-aza-CN), a well-characterized DNA methyl transferase inhibitor, to NIH-3T3 murine fibroblast cells induced the expression of stem-cell markers, that is, increasing cell plasticity. Next, 5-aza-CN treated fibroblasts were cultured in osteogenic, adipogenic, chondrogenic, and neurogenic media with or without bone morphogenetic protein 2 for a designated period. Differentiation of each desired type of cell was verified by quantitative reverse transcriptase-polymerase chain reaction/western blot assays for appropriate marker expression and by various staining methods, such as alkaline phosphatase/alizarin red S/oil red O/alcian blue. These proposed procedures allowed easier acquisition of the desired cells without any transgenic modification, using direct reprogramming technology, and thus may help make it more available in the clinical fields of regenerative medicine.

• BMB Reports
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• v.55 no.9
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• pp.447-452
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• 2022

Neurogenic differentiation 1 (NeuroD1) is an essential transcription factor for neuronal differentiation, maturation, and survival, and is associated with inflammation in lipopolysaccharide (LPS)-induced glial cells; however, the concrete mechanisms are still ambiguous. Therefore, we investigated whether NeuroD1-targeting miRNAs affect inflammation and neuronal apoptosis, as well as the underlying mechanism. First, we confirmed that miR-30a-5p and miR-153-3p, which target NeuroD1, reduced NeuroD1 expression in microglia and astrocytes. In LPS-induced microglia, miR-30a-5p and miR-153-3p suppressed pro-inflammatory cytokines, reactive oxygen species, the phosphorylation of c-Jun N-terminal kinase, extracellular-signal-regulated kinase (ERK), and p38, and the expression of cyclooxygenase and inducible nitric oxide synthase (iNOS) via the NF-κB pathway. Moreover, miR-30a-5p and miR-153-3p inhibited the expression of NOD-like receptor pyrin domain containing 3 (NLRP3) inflammasomes, NLRP3, cleaved caspase-1, and IL-1β, which are involved in the innate immune response. In LPS-induced astrocytes, miR-30a-5p and miR-153-3p reduced ERK phosphorylation and iNOS expression via the STAT-3 pathway. Notably, miR-30a-5p exerted greater anti-inflammatory effects than miR-153-3p. Together, these results indicate that miR-30a-5p and miR-153-3p inhibit MAPK/NF-κB pathway in microglia as well as ERK/STAT-3 pathway in astrocytes to reduce LPS-induced neuronal apoptosis. This study highlights the importance of NeuroD1 in microglia and astrocytes neuroinflammation and suggests that it can be regulated by miR-30a-5p and miR-153-3p.

• Tuberculosis and Respiratory Diseases
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• v.40 no.4
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• pp.345-352
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• 1993

The role of magnetic resonance(MR) imaging in the evaluation of thoracic disease has been limited Nontheless, MR has inherent properties of better contrast resolution than CT allowing tissue-specific diagnosis. MR has capability of direct imaging in sagittal, coronal, and oblique planes which provide better anatomic information than axial images of CT such as lesions in the pulmonary apex, aorticopulmonary window, peridiaphragmatic region, and subcarinal region. MR is sensitive to blood flow making it an ideal imaging modality for the evaluation of cardiovascular system of the thorax without the need for intravenous contrast media. Technical developments and better control of motion artifacts have resulted in improved image quality, and clinical applications of MR imaging in thoracic diseases have been expanded. Although MR imaging is considered as a problem-solving tool in patients with equivocal CT findings, MR should be used as the primary imaging modality in the following situations: 1) Evaluation of the cardiovascular abnormalities of the thorax 2) Evaluation of the superior sulcus tumors 3) Evaluation of the chest wall invasion or mediastinal invasion by tumor 4) Evaluation of the posterior mediastinal mass, especially neurogenic tumor 5) Differentiation of fibrosis and residual or recurrent tumor, especially in lymphoma 6) Evaluation of brachial plexopathy With technical developments and fast scan capabilities, clinical indications for MR imaging in thorax will increase in the area of pulmonary parenchymal and pulmonary vascular imaging.

• The Journal of Internal Korean Medicine
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• v.24 no.2
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• pp.380-386
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• 2003

Patients with neurogenic bladder undergo morphometric and functional changes of their bladder and urethra. As a result, voiding symptoms such as frequency, nocturia, urgency and incontinence appear. We experienced two cases of the urinary incontinence treated with Samboo-tang(Sanfu-tang). In the point of Differentiation of Syndrome, two cases were diagnosed as Deficiency Syndrome of Yang of the kidneys. After Samboo-tang(Sanfu-tang) administration, the frequency of the urinary incontinence were decreased and the accompanied symptoms also improved. So, we report two cases with a brief view of related literatures.

• Journal of Life Science
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• v.29 no.12
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• pp.1329-1336
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• 2019

Subventricular zone (SVZ) in the brain contains neural stem cells (NSCs) which self-renew and differentiate to neurons and glial cells during postnatal period and throughout adulthood. Since fate decision to either proliferation or differentiation has to respond to intracellular and extracellular conditions, many intrinsic and extrinsic factors are involved. Among them, mitochondria have been reported to participate in fate decision of NSCs. In our previous report, we showed that long-term treatment of a mitochondrial inhibitor rotenone greatly inhibited neurogenesis. In this study, we examined the effects of short-term treatment of rotenone on SVZ NSCs. We found that (1) even one-day treatment of rotenone significantly reduced neurogenesis and earlier time points seemed to be more sensitive to rotenone, (2) a number of Mash1+ transit amplifying cells was decreased by one-day treatment of rotenone, (3) short-term treatment of rotenone eliminated most of the differentiated Tuj1+ neurons and Olig2+ oligodendrocytes, while glial fibrillary acidic protein (GFAP)+ astrocytes were not affected, and (4) sulfiredoxin 1 (Srxn1) gene expression was increased after one-day treatment of rotenone, indicating activation of nuclear factor (erythroid-derived 2)-like 2 (Nrf2) pathway. All these results confirm that functional mitochondria are necessary during differentiation to neurons or oligodendrocytes as well as maintenance of neurons after differentiation. Also, these data suggest that temporary exposure to mitochondrial inhibitor such as rotenone might have long-term effects on neurogenic potential of NSCs.

• Journal of Life Science
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• v.28 no.12
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• pp.1397-1405
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• 2018

Mitochondria have multiple functions in cells: providing chemical energy, storing cellular $Ca^{2+}$, generating reactive oxygen species, and regulating apoptosis. Through these functions, mitochondria are also involved in the maintenance, proliferation, and differentiation of stem/progenitor cells. In the brain, the subventricular zone (SVZ) is one of the neurogenic regions that contains neural stem cells (NSCs) throughout a lifetime. However, reports on the role of mitochondria in SVZ NSCs are scarce. Here, we show that rotenone, a complex I inhibitor of mitochondria, inhibits the proliferation and differentiation of SVZ NSCs in different ways. In proliferating NSCs, rotenone decreases mitosis as measured through phosphorylated histone H3 detection; moreover, apoptosis is not induced by rotenone at 50 nM. In differentiating NSCs, rotenone blocks neurogenesis and oligodendrogenesis while glial fibrillary acidic protein-positive astrocytes are not affected. Interestingly, in this study there were more cells in the differentiating NSCs treated with rotenone for 4-6 days than in the vehicle control group which was a different effect from the reduced number of cells in the proliferating NSCs. We examined both apoptosis and mitosis and found that rotenone decreased apoptosis as detected by staining cleaved caspase-3 but did not affect mitosis. Our results suggest that functional mitochondria are necessary in both the proliferation and differentiation of SVZ NSCs. Furthermore, mitochondria might be involved in the mitosis and apoptosis that occur during those processes.

• Journal of Life Science
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• v.32 no.2
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• pp.108-118
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• 2022

The subventricular zone (SVZ) in the brain contains neural stem cells (NSCs) that generate new neurons throughout one's lifetime. Many extracellular and intracellular factors that affect cell proliferation and neuronal differentiation of NSCs are already well-known. Recently, L-type calcium channels have been reported to regulate neural development and are present in NSCs, differentiating neuroblasts, and mature neurons in the SVZ. Nifedipine, a blocker of L-type calcium channels, has been long used as a therapeutic drug for hypertension. However, studies on the use of nifedipine to inhibit L-type calcium channels of NSCs are lacking. Herein, we treated NSCs cultured from mouse postnatal SVZ with nifedipine during neuronal differentiation. Nifedipine increased the number of Tuj1-positive neurons but did not significantly change the number of Olig2-positive oligodendrocytes. Nifedipine increased cell division during early differentiation, which was detected using the 5-ethynyl-2'-deoxyuridine incorporation assay and immunocytochemistry assessment by staining the cells with phosphorylated histone H3, a mitosis marker. Nifedipine increased the transcription of Dlx2, a neurogenic transcription factor, and the level of Mash1, a marker for early neurogenesis. In addition to nifedipine, verapamil, which is also an L-type calcium channel blocker, showed a slight increase in neurogenesis, but its statistical significance was very low. In contrast, pimozide, a T-type calcium channel blocker, did not affect neurogenesis, although T-type calcium channel genes Cav3.1, Cav3.2, and Cav3.3 were expressed. In summary, nifedipine might promote the neuronal fate of NSCs during early differentiation and calcium signaling through L-type calcium channels might be involved in neuronal differentiation, especially during the early stages of differentiation.

• The Journal of Internal Korean Medicine
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• v.23 no.2
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• pp.286-291
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• 2002

Urinary tract infection(UTI) is one of the common complications in stroke patients. As it has negative effect on the recovery of stroke, it should be cured out immediately. But antibiotics might cause some adverse reactions such as diarrhea, eruption, anorexia, nausea and vomiting. so there have been several reports about treating urinary tract infection with Traditional Korean Medication. We treated a 54-year-old male patient with cerebral hemorrhage, who had had neurogenic bladder after stroke and had been urinated by intermittent catheterization. About 10 days later, he could void by himself without catheter, but showing the symptoms of UTI; Voiding pain, hematuria and yellowish pus. The pus culture grew Staphylococcus spp., which was resistant to most of antibiotics except vancomycin and teicoplanin. Based on accompanying symptoms of intermittent dizziness, headache, insomnia, nocturnal sweating, weak pulse, red tongue and urinary problem, we differentiated him as the deficiency of Yin of the Kidney[腎陰虛] and treated with Gagamyookmijihwang-tang (Jia-Jian-Liu-Wei-De- Huang-Tang), which improved his urinary symptoms and other general conditions without any side effect. In next follow-up culture, there was no pathogen. We conclude that Traditional Korean Medicine based on differentiation is useful in the treatment of urinary tract infection.

• The Korean Journal of Physiology and Pharmacology
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• v.22 no.2
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• pp.145-153
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• 2018

The subgranular zone (SGZ) of hippocampal dentate gyrus (HDG) is a primary site of adult neurogenesis. Toll-like receptors (TLRs), are involved in neural system development of Drosophila and innate immune response of mammals. TLR2 is expressed abundantly in neurogenic niches such as adult mammalian hippocampus. It regulates adult hippocampal neurogenesis. However, the role of TLR2 in adult neurogenesis is not well studied in global or focal cerebral ischemia. Therefore, this study aimed to investigate the role of TLR2 in adult neurogenesis after photochemically induced cerebral ischemia. At 7 days after photothrombotic ischemic injury, the number of bromodeoxyuridine (BrdU)-positive cells was increased in both TLR2 knock-out (KO) mice and wild-type (WT) mice. However, the increment rate of BrdU-positive cells was lower in TLR2 KO mice compared to that in WT mice. The number of doublecortin (DCX) and neuronal nuclei (NeuN)-positive cells in HDG was decreased after photothrombotic ischemia in TLR2 KO mice compared to that in WT mice. The survival rate of cells in HDG was decreased in TLR2 KO mice compared to that in WT mice. In contrast, the number of cleaved-caspase 3 (apoptotic marker) and the number of GFAP (glia marker)/BrdU double-positive cells in TLR2 KO mice were higher than that in WT mice. These results suggest that TLR2 can promote adult neurogenesis from neural stem cell of hippocampal dentate gyrus through increasing proliferation, differentiation, and survival from neural stem cells after ischemic injury of the brain.

• Journal of the Korean Society of Laryngology, Phoniatrics and Logopedics
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• v.28 no.2
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• pp.100-105
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• 2017

Background and Objectives : Adductor type spasmodic dysphonia (ADSD) is neurogenic disorder and focal laryngeal dystonia, while muscle tension dysphonia (MTD) is caused by functional voice disorder. Both ADSD and MTD may be associated with excessive supraglottic contraction and compensation, resulting in a strained voice quality with spastic voice breaks. The aim of this study was to determine the utility of spectrogram analysis in the differentiation of ADSD from MTD. Materials and Methods : From 2015 through 2017, 17 patients of ADSD and 20 of MTD, underwent acoustic recording and phonatory function studies, were enrolled. Jitter (frequency perturbation), Shimmer (amplitude perturbation) were obtained using MDVP (Multi-dimensional Voice Program) and GRBAS scale was used for perceptual evaluation. The two speech therapist evaluated a wide band (11,250 Hz) spectrogram by blind test using 4 scales (0-3 point) for four spectral findings, abrupt voice breaks, irregular wide spaced vertical striations, well defined formants and high frequency spectral noise. Results : Jitter, Shimmer and GRBAS were not found different between two groups with no significant correlation (p>0.05). Abrupt voice breaks and irregular wide spaced vertical striations of ADSD were significantly higher than those of MTD with strong correlation (p<0.01). High frequency spectral noise of MTD were higher than those of ADSD with strong correlation (p<0.01). Well defined formants were not found different between two groups. Conclusion : The wide band spectrograms provided visual perceptual information can differentiate ADSD from MTD. Spectrogram analysis is a useful diagnostic tool for differentiating ADSD from MTD where perceptual analysis and clinical evaluation alone are insufficient.