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Cloning and expression of lin-28 homolog B gene in the onset of puberty in Duolang sheep

  • Xing, Feng;Zhang, Chaoyang;Kong, Zhengquan
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.1
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    • pp.23-30
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    • 2019
  • Objective: Recent studies have demonstrated that lin-28 homolog B (LIN28B)/miRNA let-7 (let-7) plays a role in the regulation of pubertal onset in mammals. However, the role of LIN28B/let-7 in the onset of ovine puberty remains unknown. We cloned the Duolang sheep Lin28B cDNA sequence, detected the expression change of LIN28B, let-7a and let-7g in hypothalamus, pituitary and ovary tissues at three different pubertal stages. Methods: The reverse transcriptase polymerase chain reaction (RT-PCR) was used to clone the cDNA sequence of LIN28B gene from Duolang sheep and the bioinformatics methods were applied to analyze the amino acid sequence of LIN28B protein. The mRNA expression levels of the LIN28B gene at different pubertal stages were examined by real time RT-PCR. Results: LIN28B cDNA of Duolang sheep was cloned, and two transcripts were obtained. The amino acid sequence of transcript 1 shares 99.60%, 98.78%, and 94.80% identity with those of goat, wild yak and pig, respectively. Strong LIN28B mRNA expression was detected in the hypothalamus, pituitary, ovary, oviduct and uterus, while moderate expression was found in the liver, kidney, spleen and heart, weak expression was observed in the heart. No expression was found in the lungs. Quantitative real-time PCR (QPCR) and western-blot analysis revealed that the LIN28B was highly expressed in the hypothalamus and ovary at prepuberty stages, and this expression significantly decreased from the prepuberty to puberty stages (p<0.05). Markedly increased levels of mRNA expression were detected in the pituitary from prepuberty to puberty (p<0.05) and then significantly decreased from puberty to post-puberty (p<0.05). The expression levels of let-7a and let-7g showed no significant changes among different pubertal stages (p>0.05). Conclusion: These results provided a foundation for determining the functions of LIN28B/let-7 and their role in the onset of sheep puberty.

Combined application of rapamycin and atorvastatin improves lipid metabolism in apolipoprotein E-deficient mice with chronic kidney disease

  • Song, Eun Ju;Ahn, Sanghyun;Min, Seung-Kee;Ha, Jongwon;Oh, Goo Taeg
    • BMB Reports
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    • v.54 no.3
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    • pp.170-175
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    • 2021
  • Atherosclerosis arising from the pro-inflammatory conditions associated with chronic kidney disease (CKD) increases major cardiovascular morbidity and mortality. Rapamycin (RAPA) is known to inhibit atherosclerosis under CKD and non-CKD conditions, but it can cause dyslipidemia; thus, the co-application of lipid-lowering agents is recommended. Atorvastatin (ATV) has been widely used to reduce serum lipids levels, but its synergistic effect with RAPA in CKD remains unclear. Here, we analyzed the effect of their combined treatment on atherosclerosis stimulated by CKD in apolipoprotein E-deficient (ApoE-/-) mice. Oil Red O staining revealed that treatment with RAPA and RAPA+ ATV, but not ATV alone, significantly decreased the atherosclerotic lesions in the aorta and aortic sinus, compared to those seen in the control (CKD) group. The co-administration of RAPA and ATV improved the serum lipid profile and raised the expression levels of proteins involved in reverse cholesterol transport (LXRα, CYP7A1, ABCG1, PPARγ, ApoA1) in the liver. The CKD group showed increased levels of various genes encoding atherosclerosispromoting cytokines in the spleen (Tnf-α, Il-6 and Il-1β) and aorta (Tnf-α and Il-4), and these increases were attenuated by RAPA treatment. ATV and RAPA+ATV decreased the levels of Tnf-α and Il-1β in the spleen, but not in the aorta. Together, these results indicate that, in CKD-induced ApoE-/- mice, RAPA significantly reduces the development of atherosclerosis by regulating the expression of inflammatory cytokines and the co-application of ATV improves lipid metabolism.

Developing a fitted torso pattern for obese males in their forties and fifties utilizing virtual fitting (3D 가상착의를 활용한 40~50대 비만남성 피티드 토르소 패턴 개발)

  • Jeong-Ah, Jang;Ui-Jung, Kwon
    • The Research Journal of the Costume Culture
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    • v.30 no.6
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    • pp.828-841
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    • 2022
  • This study aims to develop a fitted torso pattern suitable for males in their forties and fifties who are obese-a group that accounts for more than fifty percent of men in this age group-using a 3D virtual fitting program. Size Korea's 2022 data for obese males in their forties and fifties is utilized for this study. In terms of the research method, a 3D program (CLO 3D) is used to implement the target group's various body types and to apply fitted torsos that enable verification and evaluation of the pattern's fit and ease. The characteristics of fitted torso patterns for obese bodies are as follows: the front centerline is a profile line; shoulder darts are added by reflecting waist-abdominal protrusion in the front panel; and shoulder, arm, waist, and center-back darts are constructed for the back panel. For obese bodies, the minimum circumference ease amounts should be approximately 6-7cm for the chest, 6cm for the waist, and 6-7cm for the hips. Based on an evaluation of the virtual fit of the developed fitted torso pattern, the data shows that a pattern (foundation) such as this is well suited for obese body types with an average of 4.67/5 points for reverse triangular obesity, an average of 6.67/7 points for log obesity, and an average of 6.69 points for the ease amount.

Replicative Senescence of Periodontal Fibroblasts Induces the Changes in Gene Expression Pattern

  • Yi, Tac-Ghee;Jun, Ji-Hae;Min, Byung-Moo;Kim, Moon-Kyu;Kim, Gwan-Shik;Baek, Jeong-Hwa
    • International Journal of Oral Biology
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    • v.32 no.1
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    • pp.35-43
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    • 2007
  • Tooth loss in elderly is mainly caused by alveolar bone loss via severe periodontitis. Although the severity of periodontitis is known to be affected by age, the aging process or the genetic changes during the aging of periodontal tissue cells are not well characterized. In this study, we investigated the effect of in vitro aging on the change of gene expression pattern in periodontal fibroblasts. Gingival fibroblasts (GF) and periodontal ligament fibroblasts (PDL) were obtained from two young patients and replicative senescence was induced by sequential subcultivation. When more than 90% cells were positively stained with senescence-associated ${\beta},-galactosidase$, those cells were regarded as aged cells. In aged GF and PDL, the level of phosphorylated retinoblastoma (RB) and $p16^{INK4A}$ protein was significantly decreased and increased, respectively. However, the protein level of p53 and p21, well known senescence-inducing genes, did not increase in aged GF and PDL. Although $p27^{Kip1}$ and $p15^{INK4B}$, another cyclin-dependent kinase inhibitors, were reported to be involved in replicative senescence of human cells, they were decreased in aged GF and PDL. Because senescent cells showed flattened and enlarged cell shape and are known to have increased focal adhesion, we examined the protein level of several integrins. Aged GF and PDL showed increased protein level of integrin ${\alpha}2$, ${\alpha}v$, and ${\beta}1$. When the gene expression profiles of actively proliferating young cells and aged cells were compared by cDNA microarray of 3,063 genes and were confirmed by reverse transcription-polymerase chain reaction, 7 genes and 15 genes were significantly and commonly increased and decreased, respectively, in aged GF and PDL. Among them, included are the genes that were known to be involved in the regulation of cell cycle, gene transcription, or integrin signaling. The change of gene expression pattern in GF and PDL was minimally similar to that of oral keratinocyte. These results suggest that $p16^{INK4A}/RB$ might be involved in replicative senescence of periodontal fibroblasts and the change of gene expression profile during aging process is cell type specific.

The Effects of Supungsunki-hwan on High Fat, High Carbohydrate Diet-induced Obese Type 2 Diabetic Mouse Model (수풍순기환 투여가 고지방, 고탄수화물 식이로 유발된 비만형 제2형 당뇨병 동물모델에 미치는 영향)

  • Park, Jong-Seol;Lee, Byung-Cheol;Doo, Ho-Kyung;Ahn, Young-Min;Ahn, Se-Young
    • The Journal of Internal Korean Medicine
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    • v.30 no.2
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    • pp.257-269
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    • 2009
  • Objective : Obesity is an important cause of diabetes, and lipotoxicity causes insulin resistance. Recently a lot of research is being done on PPAR-${\alpha}$. TNF-${\alpha}$. adiponectin, and leptin, which are important obesity related factors. In this study, we investigated the effects of Supungsunki-hwan on high fat. high carbohydrate diet-induced obese type 2 diabetic mouse models. Methods: Diabetes was induced in ICR male mouse (30${\pm}$5g) with Surwit's high fat, high sucrose diet. Mice were divided into 4 groups(n=10) of Normal. Control. Supungsunkj-hwan group. and acarbose group. The Supungsunki-hwsn group was given 10% Supungsunkj-hwan in their diet. and the acarbose group was given 0.5% acarbose in their diet. After 6 weeks. body weight. food intake, FBS and OGTT. lipid profile and liver enzymes, epididymal fat weight, and gene expression of leptin, adiponectin, TNF-${\alpha}$ and PPAR-${\alpha}$ were measured. Leptin. adiponectin. tumor necrosis factor(TNF)-${\alpha}$ and peroxisome proliferator-activated receptor (PPAR)-${\alpha}$ were evaluated by reverse transcription-polymerase chain reaction. Results : Supungsunkj-hwan increased the gene expression of PPAR-${\alpha}$, which reduces lipotoxicity and insulin resistance. Supungsunkj-hwan also significantly reduced triglyceride. AST. ALT serum levels. and 1 hour oral glucose tolerance levels. Conclusion : These results show that Supungsunkj-hwan improves insulin resistance in the liver and muscles, by reducing triglyceride levels and lipotoxicity through increased PPAR-${\alpha}$ gene expression. This is supported by the fact that Supungsunkj-hwan significantly reduces 1 hour oral glucose tolerance levels. Therefore we suggest that Supungsunkj-hwan would be an effective treatment for obese type 2 diabetic patients.

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Transcriptome Profiling of Kidney Tissue from FGS/kist Mice, the Korean Animal Model of Focal Segmental Glomerulosclerosis (국소성 분절성 사구체 신병증의 동물 모델 (FGS/kist 생쥐) 신 조직의 유전자 발현 양상)

  • Kang, Hee-Gyung;Lee, Byong-Sop;Lee, Chul-Ho;Ha, Il-Soo;Cheong, Hae-Il;Choi, Yong
    • Childhood Kidney Diseases
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    • v.15 no.1
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    • pp.38-48
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    • 2011
  • Purpose: Focal segmental glomerulosclerosis (FSGS) is the most common glomerulopathy causing pediatric renal failure. Since specific treatment targeting the etiology and pathophysiology of primary FSGS is yet elusive, the authors explored the pathophysiology of FSGS by transcriptome analysis of the disease using an animal model. Methods: FGS/kist strain, a mouse model of primary FSGS, and RFM/kist strain, as control and the parent strain of FGS/kist, were used. Kidney tissues were harvested and isolated renal cortex was used to extract mRNA, which was run on AB 1700 mouse microarray chip after reverse transcription to get the transcriptome profile. Results: Sixty two genes were differentially expressed in FGS/kist kidney tissue compared to the control. Those genes were related to cell cycle/cell death, immune reaction, and lipid metabolism/vasculopathy, and the key molecules of their networks were TNF, IL-6/4, IFN${\gamma}$, TP53, and PPAR${\gamma}$. Conclusion: This study confirmed that renal cell death, immune system activation with subsequent fibrosis, and lipid metabolism-related early vasculopathy were involved in the pathophysiology of FSGS. In addition, the relevance of methodology used in this study, namely transcriptome profiling, and Korean animal model of FGS/kist was validated. Further study would reveal novel pathophysiology of FSGS for new therapeutic targets.

Sedimentation and Distribution Pattern of the Fine-grained Sediments in the Southeastern Inner Shelf of Korea (한국 남동해역 내대륙붕 세립퇴적물의 분포 및 퇴적작용)

  • YOO Dong Geun;KIM Gil Young;LEE Ho Young;SEO Young Kyo;PARK Soo Chul;KIM Dae Choul
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.37 no.2
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    • pp.159-169
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    • 2004
  • Sedimentation and depositional pattern of the fine-grained sediments in the southeastern inner shelf of Korea were studied using a very high-resolution seismic profiles and sediment data. The recent mud deposits up to 45 m thick are distributed in the inner shelf forming a nearshore belt from the eastern part of Geoje Island to off the Pohang along the coast. The sediment in this area consists of homogeneous mud with mean grain size between $8.6\;to\;5.3\phi$ and does not show any distinct variability It gradually becomes finer and well sorted northeastward along the coast. Sediments normally appear as structureless massive mud but X-radiographs show that some bioturbation and faint lamination are present. The sediments accumulate at a rate of 0.18-0.44 cm/yr and the rate coincides well with the long-term (a 1000-year scale) accumulation based on very high-resolution seismic data. Distribution of wet bulk density and velocity shows a gradual increase from the southeastern part of Ulsan to off Pohang, whereas porosity shows a reverse pattern. Correlations between velocity and porosity/mean grain size are different from other regions compared, due to the difference of sediment texture and sedimentary environment. The recent shelf deposits are seismically characterized by three distinct facies: 1) well-stratified (near the river mouth), 2) semi-transparent (eastern part of Geoje Island), and 3) transparent (off Ulsan). The results suggest that fine-grained sediment derived from rivers, forming a nearshore mud belt, have been transported northeastward by the northeastward-flowing coastal current.

The Effect of Enhancer on the Penetration of Indapamide through Hairless Mouse Skin (경피흡수촉진제의 영향에 따른 인다파마이드의 피부투과)

  • Seo, Hui;Jeung, Sang-Young;Park, Ji-Seon;Shin, Byung-Cheol;Hwang, Sung-Joo;Cho, Sun-Hang
    • Journal of Pharmaceutical Investigation
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    • v.37 no.4
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    • pp.237-242
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    • 2007
  • The chemical formula of indapamide is 3-(aminosulfonyl)-4-chloro-N-(2,3-dihydro-2-methyl-1H-indol-l-yl)-benzamide, Indapamide is an oral antipertensive diuretic agent indicated for the treatment of hypertensive and edema. Indapamide inhibits carbonic anhydrase enzyme. Transdermal drug delivery systems, as compared to their corresponding classical oral or injectable dosage form counterparts, offer many advantages. The most important advantages are improved systemic bioavailability of the pharmaceutical active ingredients (PAI), because the first-pass metabolism by the liver and digestive system are avoided; and the controlled, constant drug delivery profile (that is, controlled zero-order absorption). Also of importance is the reduced dose frequency compared to the conventional oral dosage forms (that is, once-a-day, twice-a-week or once-a-week). Other benefits include longer duration of therapeutic action from a single application, and reversible action. For example, patches can be removed to reverse any adverse effects that may be caused by overdosing. In order to evaluate the effects of vehicles and penetration enhancers on skin permeation of Indapamide, the skin permeation rates of Indapamide from vehicles of different composition were determined using Franz cells fitted with excised hairless skins. Solubility of Indapamide in various solvents was investigated to select a vehicle suitable for the percutaneous absorption of Indapamide, The solvents used were Tween80, Tween20, Labrasol, Lauroglycol90 (LG90) and Peceol. Lauroglycol90 increase the permeability of indapamide approximately 3.75-fold compared with the control. Tween80, Tween20, Labrasol, Lauroglycol90 (LG90) and Peceol showed flux of $0.06ug/cm^2/hr,\;0.4ug/cm^2/hr,\;0.21ug/cm^2/hr,\;0.72ug/cm^2/hr,\;0.29ug/cm^2/hr$, respectively.

GRIM-19 Expression and Function in Human Gliomas

  • Jin, Yong-Hao;Jung, Shin;Jin, Shu-Guang;Jung, Tae-Young;Moon, Kyung-Sub;Kim, In-Young
    • Journal of Korean Neurosurgical Society
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    • v.48 no.1
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    • pp.20-30
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    • 2010
  • Objective : We determined whether the expression of GRIM-19 is correlated with pathologic types and malignant grades in gliomas, and determined the function of GRIM-19 in human gliomas. Methods : Tumor tissues were isolated and frozen at $-80^{\circ}C$ just after surgery. The tissues consisted of normal brain tissue (4), astrocytomas (2), anaplastic astrocytomas (2), oligodendrogliomas (13), anaplastic oligodendrogliomas (11), and glioblastomas (16). To profile tumor-related genes, we applied RNA differential display using a $Genefishing^{TM}$ DEG kit, and validated the tumor-related genes by reverse transcription polymerase chain reaction (RT-PCR). A human glioblastoma cell line (U343MG-A) was used for the GRIM-19 functional studies. The morphologic and cytoskeletal changes were examined via light and confocal microscopy. The migratory and invasive abilities were investigated by the simple scratch technique and Matrigel assay. The antiproliferative activity was determined by thiazolyl blue Tetrazolium bromide (MTT) assay and FACS analysis. Results : Based on RT-PCR analysis, the expression of GRIM-19 was higher in astrocytic tumors than oligodendroglial tumors. The expression of GRIM-19 was higher in high-grade tumors than low-grade tumors or normal brain tissue; glioblastomas showed the highest expression. After transfection of GRIM-19 into U343MG-A, the morphology of the sense-transfection cells became larger and more spindly. The antisensetransfection cells became smaller and rounder compared with wild type U343MG-A. The MTT assay showed that the sense-transfection cells were more sensitive to the combination of interferon-$\beta$ and retinoic acid than U343MG-A cells or antisense-transfection cells; the antiproliferative activity was related to apoptosis. Conclusion : GRIM-19 may be one of the gene profiles which regulate cell death via apoptosis in human gliomas.

Sources and Distributions of Dissolved Organic Matter by Fluorescence Method in the Northeastern Pacific Ocean (북동태평양에서 형광 기법을 이용한 용존유기물의 기원 및 분포)

  • Son, Ju-Won;Son, Seung-Kyu;Ju, Se-Jong;Kim, Kyeong-Hong;Kim, Woong-Seo;Park, Yong-Chul
    • Ocean and Polar Research
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    • v.29 no.2
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    • pp.87-99
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    • 2007
  • This study was conducted to understand the source and behavior of organic matter using the fluorescent technique (excitation-emission matrix) as a part of environmental monitoring program in the Korea manganese nodule mining site in the Northeastern Pacific Ocean. Water samples were collected at $0^{\circ},\;6^{\circ}N$, and $10.5^{\circ}N$ along $131.5^{\circ}W$ in August 2005. The concentration of total organic carbon (TOC) ranged from 58.01 to $171.93\;{\mu}M-C$. The vertical distribution of TOC was characterized as higher in the surface layer and decreased with depth. At $6^{\circ}N$, depth-integrated (from surface to 200 m depth) TOC was $337.1\;gC/m^2$, which was 1.4 times higher value than other stations. The exponential decay curve fit of vertical profile of TOC indicated that 59% of organic carbon produced by primary production in the surface layer could be decomposed by bacteria in the water column. Dissolved organic matter is generally classified into two distinctive groups based on their fluorescence characteristics using three-dimensional excitation/emission (Ex/Em) fluorescence mapping technique. One is known as biomacromolecule (BM; protein-like substance; showing max. at Ex 280/Em 330), mainly originated from biological metabolism. The other is geomacromolecule (GM; humic-like substance; showing max. at Ex 330/Em 430), mainly originated from microbial degradation processes. The concentration of BM and GM was from 0.42 to 7.29 TU (tryptophan unit) and from 0.06 to 1.81 QSU (quinine sulfate unit), respectively. The vertical distribution of BM was similar to that of TOC as high in the surface and decreased with depth. However, the vertical distribution of GM showed the reverse pattern of that of BM. From these results, it appeared that BM occupied a major part of TOC and was rapidly consumed by bacteria in the surface layer. GM was mainly transformed from BM by microbial processes and was a dominant component of TOC in the deep-sea layer.