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HeLa E-Box Binding Protein, HEB, Inhibits Promoter Activity of the Lysophosphatidic Acid Receptor Gene Lpar1 in Neocortical Neuroblast Cells

  • Kim, Nam-Ho;Sadra, Ali;Park, Hee-Young;Oh, Sung-Min;Chun, Jerold;Yoon, Jeong Kyo;Huh, Sung-Oh
    • Molecules and Cells
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    • v.42 no.2
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    • pp.123-134
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    • 2019
  • Lysophosphatidic acid (LPA) is an endogenous lysophospholipid with signaling properties outside of the cell and it signals through specific G protein-coupled receptors, known as $LPA_{1-6}$. For one of its receptors, $LPA_1$ (gene name Lpar1), details on the cis-acting elements for transcriptional control have not been defined. Using 5'RACE analysis, we report the identification of an alternative transcription start site of mouse Lpar1 and characterize approximately 3,500 bp of non-coding flanking sequence 5' of mouse Lpar1 gene for promoter activity. Transient transfection of cells derived from mouse neocortical neuroblasts with constructs from the 5' regions of mouse Lpar1 gene revealed the region between -248 to +225 serving as the basal promoter for Lpar1. This region also lacks a TATA box. For the region between -761 to -248, a negative regulatory element affected the basal expression of Lpar1. This region has three E-box sequences and mutagenesis of these E-boxes, followed by transient expression, demonstrated that two of the E-boxes act as negative modulators of Lpar1. One of these E-box sequences bound the HeLa E-box binding protein (HEB), and modulation of HEB levels in the transfected cells regulated the transcription of the reporter gene. Based on our data, we propose that HEB may be required for a proper regulation of Lpar1 expression in the embryonic neocortical neuroblast cells and to affect its function in both normal brain development and disease settings.

Modified Tei index in patients with Kawasaki disease by tissue doppler imaging (가와사키병 환아에서 Tissue doppler imaging으로 측정한 modified Tei 지수)

  • Kim, Hee Jung;Cha, Jung Hwa;Hong, Young Mi
    • Clinical and Experimental Pediatrics
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    • v.49 no.11
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    • pp.1202-1210
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    • 2006
  • Purpose : A quantitative and easily measured Doppler index of combined systolic and diastolic ventricular myocardial performance (Tei index) was recently proposed as a potentially useful predictor of global myocardial performance. However, presence of heart rate fluctuation makes it unreliable. Therefore, the modified Tei index was introduced by using tissue Doppler imaging (TDI) which enables measuring contraction and relaxation velocities from myocardium simultaneously. The purpose of this study was to investigate the effectiveness of the modified Tei index in the evaluation of global cardiac function. Methods : Forty eight patients in the acute phase of Kawasaki disease (KD) were studied. These patients were divided into two groups according to the modified Tei index. TDI and conventional echocardiography were performed. Systolic velocity, systolic displacement, E' velocity, E' displacement, A' velocity and A' displacement were measured at the base, mid-septum and the apex of the interventricular septum. Ejection fraction (EF), Tei index and modified Tei index were estimated. Results : Modified Tei index correlated negatively with systolic displacement, E' displacement and A' displacement at the base. Modified Tei index correlated positively with ESR and CRP. EF and Tei index were normal in KD patients. Conclusion : Modified Tei index is a safe, feasible and sensitive index for evaluating global ventricular functions in spite of normal EF and Tei index in KD.

Effect of administration with fat-free diet and dietary iron on polyunsaturated fatty acid, vitamin E and their derivatives in male rats (Rat에 있어서 탈지사료 및 철분투여가 다불포화 지방산, vitamin E 및 그 유도체에 미치는 영향)

  • Huh, Rhin-sou;Kim, Young-hong;Kim, Sung-hoon;Do, Jae-cheul;Lee, Young-ho;Choi, Yeon-shik
    • Korean Journal of Veterinary Research
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    • v.30 no.4
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    • pp.407-412
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    • 1990
  • This study was designed to elucidate the effects of administration with dietary iron and fat-free diet on the contents of unsaturated fatty acid in phospholipid molecules, vitamin E contents and malondialdehyde contents in liver, kidney, muscle and testis of the male rats. The rats were divided into 3 experimental groups, namely, control, iron injection and fat-free diet administration groups. The control group was fed with normal diet, iron injection group injected intraperitoneally 20mg of ferric hydroxide/100g of body weight 20 times every 3 days and fat-free diet group administered lipid extraction diet with hexane in normal diet. All experimental groups were maintained for 60 days with feeding on the respective ration. The results obtained were summarized as follows: 1. In the mean contents of unsaturated fatty acid in phospholipid of liver, kidney, muscle and testis among groups, control group was 21.31mg/g, 19.38mg/g, 1.67mg/g, 13.68mg/g, iron injection group was 13.83mg/g, 16.53mg/g, 0.71mg/g, 10.11mg/g and fat-free diet group was 21.07mg/g, 19.38mg/g, 1.49mg/g and 13.40mg/g, respectively. 2. In the mean contents of vitamin E in liver, kidney, muscle and testis among groups, control group was 6.77mg/g, 1.93mg/g, 0.12mg/g, 0.17mg/g, iron injection group was 3.16mg/g, 0.86mg/g, 0.07mg/g, 0.09mg/g and fat-free diet group was 7.41mg/g, 1.50mg/g, 0.11mg/g and 0.16mg/g, respectively. 3. In the mean contents of malondialdehyde in liver, kidney, muscle, testis and serum among groups, control group was 11.29nM/0.1g, 23.25nM/0.1g, 42.47nM/0.1g, 7.01nM/0.1g, 4.33nM/ml, iron injection group was 34.98nM/0.1g, 40.55nM/0.1g, 72.21nM/0.1g, 12.26nM/0.1g, 11.27nM/ml and fat-free diet group was 8.07nM/0.1g, 20.63nM/0.1g, 39.92nM/0.1g, 6.95nM/0.1g and 4.27nM/ml, respectively.

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The Experimental Study on the Immuno-regulatory effect of Notopterygii Rhizoma Herbal-acupuncture at Pyesu(BL13) on OVA-induced asthma in mice (폐유(肺兪) 강활약침(羌活藥鍼)이 OVA-induced Asthma Mouse Model의 면역조절(免疫調節)에 미치는 영향)

  • Jang, Suk-Geun;Hong, Kwon-Eui;Lee, Byung-Ryul
    • Korean Journal of Acupuncture
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    • v.22 no.2
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    • pp.107-125
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    • 2005
  • Objectives : The aim of this study was to investigate the asthma-suppressive and immuno-regulatory effect of NR-HA(Notopterygii Rhizoma Herbal-acupuncture) at Pyesu(BL13) on OVA(ovalbumin)-induced asthma in mice. Methods : C57BL/6 mice out of all the experimental sloops, except the Normal group and the NR-HA group, were sensitized and challenged with OVA. The mice in the NR-HA group and the OVA-NR-HA group were treated with NR-HA(1%) at Pyesu(BL13). The mice in the OVA-Saline group were injected with saline at Pyesu(BL13). The mice in the OVA-Needle-prick group were treated with a single prick with an injection needle at Pyesu(BL13). NR-HA, saline injection and needle prick were administered for 8 weeks, three times a week Results : in vitro 1. The populations of granulocytes, $CD3e^-/CCR3^+$cells, $CD69^+/CD3e^+$ cells, $CD4^+\;cells\;and\;CD23^+/B220^+$ cells in the OVA-induced asthmatic mouse lungs decreased significantly by NR-HAS(Notopterygii Rhizoma Herbal-acupuncture solution). 2. The lung weight and total cells in lung of the OVA-NR-HA group decreased significantly compared with those of the OVA-Control group. 3. Total leukocytes and eosinophils in BALF of the OVA-NR-HA group decreas ed significantly compared with those of the OVA-Control group. 4. The collagen accumulation in the lung sections of the OVA-NR-HA group decreased significantly compared with that of the OVA-control group. 5. The concentrations of IL-4, IL-5, IL-13, IgE in BALF and serum of the OVA-NR-HA group decreased significantly compared with those of the OVA- Control group. 6. The numbers of $Gr-1^+/CD11b^+,\;CCR3^+,\;CD3e^+, \;CD19^+,\;CD3e^+/CD69^+$ cells in the OVA-NR-HA group decreased significantly compared with those of the OVA-Control group. 7. The mRNA expressions of $TNF-{\alpha}$, IL-5, IL-4 and IL-13 in lung of the OVA-NR-HA group decreased significantly compared with those of the OVA- Control group. 8. The NR-HA group did not show my considerable difference from the Normal group. The OVA-saline group and the OVA-Needle prick group showed suppressive effects on OVA-induced asthma however they were not statistically significant. Conclusion : These results suggest that NR-HA at Pyesu(BL13) is considered to be effective in treating asthma and to be put to practical use in the future asthma clinic.

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Effects of Acupuncture at ST41, BL60, GB38 on Changes of Nitric Oxide and Nitric Oxide Synthase in Rats (백서의 경혈(해계, 곤륜, 양보) 침자가 Nitric Oxide Synthase 및 Nitric Oxide의 변화에 미치는 영향)

  • Kim, Youngsun;Choi, Donghee;Jang, Hosun;Na, Changsu;Hwang, Moonhyeon;Cho, Joohyun;Lee, Kyoungin;Kim, Sunmin;Pyo, Byoungsik;Youn, Daehwan
    • Korean Journal of Acupuncture
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    • v.30 no.2
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    • pp.97-103
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    • 2013
  • Objectives : To observe the changes in the expression of nNOS, iNOS, eNOS and NO by the needle insertion on river points, one of the five transport points of three yang meridians of the foot. Methods : Based on rats, needle was inserted on both left and right sides of ST41, BL60 and GB38 and retained for five minutes. After the retention, blood was drawn via cardiac puncture and tissues from each point around meridian vessels were extracted to observe the changes in the expression of nNOS, iNOS, eNOS and NO. Results : In terms of the effect on expression of nNOS within tissues, ST41 showed significant decrease based on the results of immunohistochemistry. In terms of the effect on expression of iNOS within tissues, none of the experimental groups showed any significant change compared to the Normal group. Regarding expression of eNOS within tissues, GB38 showed significant increase based on the results of immunohistochemistry. In terms of the effect on NO creation in tissues, none of the experimental groups showed any significant change compared to the Normal group. Conclusions : The effect of needle insertion on the river points of three yang meridians of the foot on the function of nNOS, iNOS, eNOS and NO could be observed, and based on this study, it is considered that the effect of needle stimulation on the changes of nervous system could be found out through additional research.

Bioactive Polyglycolic Acid (PGA) or Polylactic Acid (PLA) Polymers on Extracellular Matrix Mineralization in Osteoblast-like Mc3T3-E1 Cells

  • Cho, Young-Eun;Kim, Hye-Jin;Kim, Yong-Ha;Choi, Jae-Won;Kim, Youn-Jung;Kim, Gab-Joong;Kim, Jin-Su;Choi, Sik-Young;Kwun, In-Sook
    • Nutritional Sciences
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    • v.9 no.4
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    • pp.233-239
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    • 2006
  • Porous matrices of bioactive polymers such as polyglycolic acid (PGA) or polylactic acid (PLA) can be used as scaffolds in bone tissue growth during bone repair process. These polymers are highly porous and serve as a template for the growth and organization of new bone tissues. We evaluated the effect of PGA and PLA polymers on osteoblastic MC3T3-E1 cell extracellular mineralization. MC3T3-E1 cells were cultured in a time-dependent manner -1, 15, 25d as appropriate - for the period of bone formation stages in one of the five culture circumstances, such as normal osteogenic differentiation medium, PGA-plated, fetal bovine serum (FBS)-plated, PGA/FBS-coplated, and PLA-plated For the evaluation of bone formation, minerals (Ca, Mg, Mn) and alkaline phosphatase activity, a marker for osteoblast differentiation, were measured Alizarin Red staining was used for the measurement of extracellular matrix Ca deposit During the culture period, PGA-plated one was reabsorbed into the medium more easily and faster than the PLA-plated one. At day 15, at the middle stage of bone formation, cellular Ca and Mg levels showed higher tendency in PGA- or PLA-plated treatments compared to non-plated control and at day 25, at the early late stage of bone formation, all three cellular Ca, Mg or Mn levels showed higher tendency as in order of PGA-related treatments and PLA-plated treatments, compared to control even without significance. Medium Ca, Mg or Mn levels didn't show any consistent tendency. Cellular ALP activity was higher in the PGA- or PLA-plated treatments compare to normal osteogenic medium treatment PGA-plated and PGA/FBS-plated treatments showed better Ca deposits than other treatments by measurement of Alizarin Red staining, although PLA-plated treatment also showed reasonable Ca deposit. The results of the present study suggest that biodegradable material, PGA and also with less extent for PLA, can be used as a biomaterial for better extracellular matrix mineralization in osteoblastic MC3T3-E1 cells.

A Study on Wound Healing Factors in Diabetic Foot Ulcer Patients (당뇨발 궤양을 가진 환자에서의 창상치유인자에 대한 조사)

  • Gu, Ja Hea;Han, Seung-Kyu;Chang, Hyun;Lee, Byoung Il;Kim, Woo Kyung
    • Archives of Plastic Surgery
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    • v.34 no.5
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    • pp.543-550
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    • 2007
  • Purpose: Diabetic foot ulcer is a representative disease of chronic would with multiple defects of wound healing factors. Many nutrition factors have been known to be essential for wound healing, but objective data are lacking. The purpose of this study is to determine those factors essential for wound healing, and to find out which of those factors are lacking in diabetic foot ulcer patients through this pilot study. Methods: We studied 100 patients who visited our clinic from March 2005 to February 2006 for treatment of diabetic foot ulcers with a duration of more than 6 weeks. We checked serum levels of protein, albumin, vitamin A, C, E, iron, magnesium, zinc, copper and hemoglobin by drawing 23cc blood after 8 hours of fasting. Protein, albumin, iron, magnesium levels were measured by colorimetry; hemoglobin levels were measured by auto analysis. Vitamin levels were measured by high performance liquid chromatography (HPLC), copper and zinc levels were measured by Inductively coupled plasma (ICP). They were compared with normal values. The patients were divided by transcutaneous oxygen pressure levels, age and sex to study the effects of these parameters. Results: 76% and 61% of patients had within-normal range serum protein and albumin levels, respectively. Among vitamins, only the level of vitamin C was low in 55% of the patients. Levels of vitamin A, E were normal or high in 93% and 100% of patients. As for trace elements, levels of iron and zinc were low in 63% and 60% of patients, but levels of magnesium and copper were usually normal or high. Levels of vitamin C, iron and zinc were lower in the low-transcutaneous oxygen pressure group. There were no definite differences according to age and sex. Conclusion: Serum levels of Hb, vitamin C, iron, zinc were low in most diabetic foot ulcer patients. The deficit was very severe in the low-transcutaneous oxygen pressure group.

Expression of Nuclear Factor Erythroid 2 Protein in Malignant Cutaneous Tumors

  • Choi, Chang Yong;Kim, Jin Young;Wee, Seo Yeong;Lee, Jang Hyun;Nam, Doo Hyun;Kim, Chul Han;Cho, Moon Kyun;Lee, Yoon Jin;Nam, Hae Seon;Lee, Sang Han;Ch, Sung Woo
    • Archives of Plastic Surgery
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    • v.41 no.6
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    • pp.654-660
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    • 2014
  • Background Reactive oxygen species (ROS) damages cell molecules, and modifies cell signaling. The nuclear factor E2-related factor (Nrf2) is a critical transcription regulator, which protects cells against oxidative damage. Nrf2 expression is increased in a large number of cancers. However, little information has been reported regarding the expression of Nrf2 in skin cancers. Hence, we explored the expression of Nrf2 protein in skin cancers. Methods The Nrf2 protein expression in 24 specimens, including 6 malignant melanomas (MM), 6 squamous cell carcinomas (SCC), 6 basal cell carcinomas (BCC), and 6 normal skin tissues, was evaluated by western blotting. Immunohistochemical staining was performed. The expression of Kelch-like ECH-associated protein 1 (Keap1), the key regulator of Nrf2, was also analyzed by western blotting. Results Small interfering RNA transfection to the melanoma cell line G361 confirmed that an approximately 66 kDa band was the true Nrf2 band. The western blot revealed that the Nrf2 protein was definitely expressed in normal skin tissues, but the Nrf2 expression was decreased in MM, SCC, and BCC. Immunohistochemical examination showed that expression of Nrf2 was decreased in all skin cancer tissues compared to the normal skin tissues. Keap1 was not expressed in all malignant skin tumors and normal skin tissues by western blot. Conclusions ROS was increased in various types of cancers which proteins were highly expressed or underexpressed. This study demonstrated that the expression of Nrf2 protein was down-regulated in human malignant skin tumors. We suggest that decreased expression of Nrf2 is related to skin cancers.

A Study on the Effect of Herbal-acupuncture with Carthami Flos at Joksamni($ST_{36}$) on Collagen-induced Arthritis in Mice (족삼리(足三里) 홍화약침(紅花藥鍼)처치가 Collagen으로 유발한 생쥐의 관절염 모델에 미치는 영향)

  • Park, Ki-Hong;Lee, Hyun
    • Journal of Acupuncture Research
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    • v.24 no.6
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    • pp.45-61
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    • 2007
  • Objectives : The purpose of this study is to observe the effects of Carthami Flos herbal-acupuncture (CF-HA) at Joksamni($ST_{36}$) on arthritis in mice induced by Collagen II. Methods : The author performed several experimental items, including arthritis evaluation, change in weight, spleen size and stenosis rate, change in cytokine level, IgG, IgM and anti-collagen II, change of immunocyte count and histological change of the CIA mouse joint. Conclusions are as follows: Results : 1. In the CF-HA, the arthritis index and rate and the incidence of arthritis were decreased as the experiment proceeded. 2. In the CF-HA, spleen swell and stenosis, joint edema and change were decreased. 3. In the CF-HA, the level of $IL-1{\beta}$, IL-6, $TNF-{\alpha}$ and $IFN-{\gamma}$ in blood serum were significantly decreased. 4. In the CF-HA, the level of IgG, IgM and anti-collagen II were decreased. 5. In the CF- HA, $IFN-{\gamma}$, $IFN-{\gamma}/IL-4$, IL-10 of the culture fluid was decreased. 6. In the CF-HA, the cell rate of $CD3e^+$ and $CD45R^+$, $CD4^+$ and $CD8^+$, $CD4^+/CD25^+$ in spleen was similar to the cell rate of the normal group. 7. In the CF-HA, the cell rate of $CD4^+/CD25^+$, $CD45R^+/CD69^+$ in a lymph node was decreased as in the normal group. 8. In the CF-HA, the cell rate of $CD3^+/CD69^+$, $CD11b^+/Gr-1^+$ in joints was decreased as in the normal group. 9. In the CF-HA, the cartilage destruction and the inflammation cell growth in the H&E stain were decreased. The collagen fiber in the M&T stain were less destructed, therefore the result was similar to the normal group. Conclusions : These results suggest that CH-HA at $ST_{36}$ has an effect in controlling immune reaction and suppressing inflammation in rheumatoid arthritis therefore, the continuous flow of the following study is expected.

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Effect of Tenebrio molitor Larvae Fermented with Saccharomyces cerevisiae M1 (KACC 93023) on Non-alcoholic Fatty Liver-induced Rats (Saccharomyces cerevisiae M1 (KACC 93023)으로 발효한 갈색 거저리 유충이 비 알코올성 지방간 유발 흰 쥐에 미치는 영향)

  • Sim, So-Yeon;Jang, Sung-Ho;Cho, Young-Su;Ahn, Hee-Young
    • Journal of Life Science
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    • v.30 no.5
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    • pp.434-442
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    • 2020
  • In this study, Tenebrio molitor (brown mealworm) larvae powder was fermented with Saccharomyces cerevisiae M1 (KACC 93023). The fermented and non-fermented powders were subsequently administered to rats with induced non-alcoholic fatty liver disease to compare the degree of improvement. The rats exhibited abnormal lipid metabolism due to orotic acid, and the group that were fed the fermented larvae powder showed similar weight to normal rats. AST, ALT, ALP, and LDH activities, all known indicators of liver health, and various other lipid-related indicators appeared positive, suggesting improvement. The lipid peroxide and glutathione content in the liver tissue and serum were similar to or better than those of the normal group. Finally, morphological observations of liver tissue using H&E and oil red O stainings revealed that the fermented powder group showed markedly reduced fat along with normal hepatocyte shape and arrangement. To summarize, it was confirmed that dietary brown mealworm larvae improve non-alcoholic fatty liver function, and, when fermented with yeast, most indicators showed positive improvements to almost normal levels. As such, the fermented powder can be considered to improve non-alcoholic fatty liver, suggesting that it may be used as a material for various products in the future.