• Title/Summary/Keyword: Stem cells

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Preparation and Characterization of PLGA Scaffold Impregnated Keratin for Tissue Engineering Application (케라틴이 함유된 조직공학적 PLGA 지지체의 제조 및 특성 분석)

  • Oh, A-Young;Kim, Soon-Hee;Lee, Sang-Jin;Yoo, James J.;Dyke, Mark van;Rhee, John M.;Khang, Gil-Son
    • Polymer(Korea)
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    • v.32 no.5
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    • pp.403-408
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    • 2008
  • Keratin is the major structural fibrous protein providing outer covering such as wool, hair, and nail. Keratin is useful as natural protein. We developed the keratin loaded poly(L-lactide-co-glycolide) (PLGA) scaffolds (keratin/PLGA) for the possibility of the application of the tissue engineering using bone marrow mesenchymal (BMSCs). Keratin/PLGA (contents 0%, 10%, 20% and 50% of PLGA weight) scaffolds were prepared by solvent casting/salt leaching method. We characterized porosity, wettability, and water uptake ability, DSC of keratin/PLGA scaffold. We seeded BMSCs isolated from the femurs of rat into the inner core of the hybrid scaffold. Celluar viability were assayed by 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl-tetrazolium bromide (MTT) test. We confirmed that keratin/PLGA scaffold is hydrophilic by wettability, and water uptake ability measurement results. In MTT assay results, cell viability in scaffolds impregnated 10 and 20 wt% of keratin were higher than other scaffolds. In conclusion, we suggest that keratin/PLGA scaffold may be useful to tissue engineering using BMSCs.

Ameliorating Effects of Nokyongdaebo-tang on Experimental Subacute Hemorrhagic Anemia in Rats (녹용대보탕 열수 추출물의 실험적으로 유발된 랫트 아급성 출혈성 빈혈에 대한 효과)

  • Kim, Jung-Ah;Kim, Dong-Chul
    • The Journal of Korean Obstetrics and Gynecology
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    • v.30 no.3
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    • pp.1-19
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    • 2017
  • Objectives: The object of this study is to observe the possible ameliorating effects of Nokyongdaebo-tang (NYDBT) on the experimental subacute hemorrhagic anemia (SHA) in rats. Methods: In the present study, SHA in rats was induced by exsanguinations from orbital plexus, and ameliorating effects of NYDBT was observed based on the changes of body and hematopoietic organ (spleen, liver and femur) weights, red blood cell (RBC) related hematological values, smear cytology, histopathological changes and immunohistochemistrical analysis of hematopoietic stem cells in the femur bone marrow, liver and spleen. In addition, the gastrointestinal motility and the surface mucosa thicknesses of remnant fecal pellets in the colon lumen, mucosa thicknesses and the mucous producing cell numbers in the colonic mucosa were analyzed to observe the digestive disorders, especially on the constipation, the major discomfort problems in iron supplement. Results: SHA related abnormal anemic signs were markedly and dose-dependently inhibited by oral administration of NYDBT 500, 250 and 125 mg/kg in a condition of this experiment. In addition, no meaningful changes on the gastrointestinal motilities and mucous component on the colon and remnant feces were noticed in all three different dosages of NYDBT treated rats as compared with intact vehicle and SHA control rats in this study. Conclusions: It, therefore, is expected that NYDBT will be promising as a novel alternative hematopoietic and therapeutic agent for anemia.

Manufacture and Characterization of Silkworm Gland Hydrolysate (누에 실샘 가수분해물의 제조 및 특성 규명)

  • Hwang, Jung Wook;Lee, Heui Sam;Kim, Hojin;Kim, Kyu-Oh;Choi, Yong-Soo
    • Journal of Sericultural and Entomological Science
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    • v.50 no.2
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    • pp.76-81
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    • 2012
  • Silk protein has been explored to be used for biomedical applications for several decades. However, it has not been used in this field cause to their irreversible crystallization after dissolving in water. The existing methods of silk protein hydrolysis using silkworm cocoon were used with harmful solvents and through a very complicated process. Therefore, we have developed novel methods for the production of water-soluble hydrolysate using silkworm gland. We manufactured two types of silkworm gland-derived hydrolysate (water-soluble SGH, SSGH; total SGH, TSGH) and compared the characteristics with commercial cocoon-derived sericin hydrolysate (CSH). The molecular weight of SGH ranged from 7 to 50 kDa (SSGH) and 5 to 15 kDa (TSGH) within glycine, alanine, and aspartic acid as a main amino acid composition. In contrast, CSH ranged from 15 to 50 kDa within serine and aspartic acid. The results of FTIR implied that SGH was more soluble form than CSH, as shown by the decrease in the ${\beta}$-sheet structure at $1630cm^{-1}$ on amide I peak. In comparison with 10% fetal bovine serum, 0.1% (1 mg/ml) SSGH had equivalent effect on the proliferation of human dermal fibroblasts and mesenchymal stem cells. All results of the SSGH made by novel manufacturing process indicate the SSGH is more preferable as a culture medium supplement than cocoon-derived sericin.

Effect of an Endoplasmic Reticulum Retention Signal Tagged to Human Anti-Rabies mAb SO57 on Its Expression in Arabidopsis and Plant Growth

  • Song, Ilchan;Lee, Young Koung;Kim, Jin Wook;Lee, Seung-Won;Park, Se Ra;Lee, Hae Kyung;Oh, Soyeon;Ko, Kinarm;Kim, Mi Kyung;Park, Soon Ju;Kim, Dae Heon;Kim, Moon-Soo;Kim, Do Sun;Ko, Kisung
    • Molecules and Cells
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    • v.44 no.10
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    • pp.770-779
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    • 2021
  • Transgenic Arabidopsis thaliana expressing an anti-rabies monoclonal antibody (mAb), SO57, was obtained using Agrobacterium-mediated floral dip transformation. The endoplasmic reticulum (ER) retention signal Lys-Asp-Glu-Leu (KDEL) was tagged to the C-terminus of the anti-rabies mAb heavy chain to localize the mAb to the ER and enhance its accumulation. When the inaccurately folded proteins accumulated in the ER exceed its storage capacity, it results in stress that can affect plant development and growth. We generated T1 transformants and obtained homozygous T3 seeds from transgenic Arabidopsis to investigate the effect of KDEL on plant growth. The germination rate did not significantly differ between plants expressing mAb SO57 without KDEL (SO plant) and mAb SO57 with KDEL (SOK plant). The primary roots of SOK agar media grown plants were slightly shorter than those of SO plants. Transcriptomic analysis showed that expression of all 11 ER stress-related genes were not significantly changed in SOK plants relative to SO plants. SOK plants showed approximately three-fold higher mAb expression levels than those of SO plants. Consequently, the purified mAb amount per unit of SOK plant biomass was approximately three times higher than that of SO plants. A neutralization assay revealed that both plants exhibited efficient rapid fluorescent focus inhibition test values against the rabies virus relative to commercially available human rabies immunoglobulins. KDEL did not upregulate ER stress-related genes; therefore, the enhanced production of the mAb did not affect plant growth. Thus, KDEL fusion is recommended for enhancing mAb production in plant systems.

Antioxidant and Anti-Inflammatory Activities of Ethanol Extract of Clematis trichotoma Nakai (할미밀망 에탄올 추출물의 항산화 및 항염증 활성 평가)

  • Jung, Jaemee;Shin, Mijoon;Jeong, Naeun;Hwang, Dahyun
    • Korean Journal of Clinical Laboratory Science
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    • v.53 no.2
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    • pp.165-173
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    • 2021
  • Clematis trichotoma Nakai (CTN) is a broad-leaved vine plant belonging to the family Ranunculus, native to Korea. Young leaves are used as food, and the stem and roots are used as medicinal materials. Antioxidant studies have been reported on the stems of CTN, but no studies have been conducted on the leaves. In this study, a 70% ethanol extract of CTN was prepared and its antioxidant and anti-inflammatory activities were investigated. For measuring the antioxidant activity, five assays (polyphenol and flavonoid content, reducing power, 2,2-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid), and 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity) were performed and CTN showed a concentration-dependent effect in all assays. To investigate the anti-inflammatory activity, we used RAW 264.7 cells. The concentrations (from 31.25 to 250 ㎍/mL) of CTN did not show cytotoxicity. CTN (250 ㎍/mL) inhibited dendritic transformation (34.4%) and also inhibited inflammation as seen by reduced levels of NO (77.4%), IL-6 (85.5%) and TNF-α (41.2%) compared to lipopolysaccharide (LPS). CTN (250 ㎍/mL) also suppressed the expression of the following genes: COX-2 (79.8%), iNOS2 (93.9%), IL-6 (87.6%), and TNF-α (77.3%) compared to LPS. These results demonstrated that CTN has excellent antioxidant and anti-inflammatory activities and can therefore be used as a natural biological resource.

20(S)-protopanaxadiol and oleanolic acid ameliorate cognitive deficits in APP/PS1 transgenic mice by enhancing hippocampal neurogenesis

  • Lin, Kaili;Sze, Stephen Cho-Wing;Liu, Bin;Zhang, Zhang;Zhang, Zhu;Zhu, Peili;Wang, Ying;Deng, Qiudi;Yung, Ken Kin-Lam;Zhang, Shiqing
    • Journal of Ginseng Research
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    • v.45 no.2
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    • pp.325-333
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    • 2021
  • Background: Alzheimer's disease (AD) is one of the most prevalent neurodegenerative disorders. Enhancing hippocampal neurogenesis by promoting proliferation and differentiation of neural stem cells (NSCs) is a promising therapeutic strategy for AD. 20(S)-protopanaxadiol (PPD) and oleanolic acid (OA) are small, bioactive compounds found in ginseng that can promote NSC proliferation and neural differentiation in vitro. However, it is currently unknown whether PPD or OA can attenuate cognitive deficits by enhancing hippocampal neurogenesis in vivo in a transgenic APP/PS1 AD mouse model. Here, we administered PPD or OA to APP/PS1 mice and monitored the effects on cognition and hippocampal neurogenesis. Methods: We used the Morris water maze, Y maze, and open field tests to compare the cognitive capacities of treated and untreated APP/PS1 mice. We investigated hippocampal neurogenesis using Nissl staining and BrdU/NeuN double labeling. NSC proliferation was quantified by Sox2 labeling of the hippocampal dentate gyrus. We used western blotting to determine the effects of PPD and OA on Wnt/GSK3β/β-catenin pathway activation in the hippocampus. Results: Both PPD and OA significantly ameliorated the cognitive impairments observed in untreated APP/PS1 mice. Furthermore, PPD and OA significantly promoted hippocampal neurogenesis and NSC proliferation. At the mechanistic level, PPD and OA treatments resulted in Wnt/GSK-3β/β-catenin pathway activation in the hippocampus. Conclusion: PPD and OA ameliorate cognitive deficits in APP/PS1 mice by enhancing hippocampal neurogenesis, achieved by stimulating the Wnt/GSK-3β/β-catenin pathway. As such, PPD and OA are promising novel therapeutic agents for the treatment of AD and other neurodegenerative diseases.

Current Trends in the Treatment of Osteochondral Lesion of the Talus: Analysis of the Korean Foot and Ankle Society (KFAS) Member Survey (거골 골연골병변 치료 동향: 대한족부족관절학회 회원 설문조사 분석)

  • Cho, Byung-Ki;Cho, Jaeho;Young, Ki Won;Lee, Dong Yeon;Bae, Su-Young;The Academic Committee of Korean Foot and Ankle Society,
    • Journal of Korean Foot and Ankle Society
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    • v.25 no.4
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    • pp.149-156
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    • 2021
  • Purpose: Given the lack of definite evidence-based guidelines in clinical practice, there may be a wide variation in treatment protocols for osteochondral lesions of the talus (OLT). Based on the Korean Foot and Ankle Society (KFAS) member survey, this study aimed to report the current trends in the management of OLT. Materials and Methods: A web-based questionnaire containing 30 questions was sent to all KFAS members in September 2021. The questions were mainly related to clinical experience and preferences in diagnosis, conservative, and surgical treatments for patients with OLT. Answers with a prevalence of ≥50% of respondents were considered a tendency. Results: Sixty-two (11.3%) of the 550 surgeons queried responded to the survey. The responses to 9 (30.0%) of the total of 30 questions established a tendency. Answers exhibiting a tendency were as follows; additional diagnostic tools except for plain radiograph (magnetic resonance imaging), most common conservative treatment method (oral medication, rest), most important radiological factor in decision making for surgical treatment and method (size of the lesion, ankle instability, loose bodies), most important patient factors in decision making for surgical treatment and method (age, activity or occupation), infrequently requiring posterior arthroscopy (less than 3%), most common revision surgery for failed bone marrow stimulation procedure (osteochondral autograft transplantation [OAT]), not requiring additional procedure for donor site in OAT, the main reason for unsatisfactory result after OAT (persistent pain without radiological abnormality), no generalization of autologous chondrocyte implantation or chondrogenesis using stem cells. Conclusion: This study presents updated information on current trends in the management of OLT in Korea. Both consensus and variations in the approach to patients with OLT were revealed through this survey. Since recent biologic efforts to regenerate cartilage have been unsuccessful, further studies to identify clinical evidence would be needed.

Study of the Function of β-D-fructofuranosidase by Plant Histological Method (식물 조직학적 방법에 의한 β-D-fructofuranosidase의 기능 연구)

  • Donggiun Kim
    • The Journal of the Convergence on Culture Technology
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    • v.9 no.4
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    • pp.587-592
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    • 2023
  • Plant growth is regulated by a variety of factors, including organic matter availability. Organic nutrients are carbohydrate molecules from photosynthetic products produced by tissues associated with carbon and energy fixation called "sources". These compounds flow through plant vascular bundles into non-photosynthetic or growing tissues called "sinks". Among these possible compounds, the disaccharide fructosyl glucose, sucrose, is the most representative. During the transport of sucrose, the pathway from the source to the sinks can include hydrolysis of sucrose into glucose and fructose derivatives or direct transfer of sucrose. Among the enzymes involved in this, β-D-fructofuranosidase is the most important. Soluble neutral β-D-fructofuranosidase, one of several isoenzymes, is located in intracellular protoplasts and helps plant cells metabolize sucrose to produce energy. In order to track the activity of this enzyme during the course of plant growth, histological methods were used for the most effective immunolocalization. As a result, the activity was higher in the phloem and epidermis than in the mesophyll tissue in the leaf. In the growing stem, activity was high in the phloem, epidermis, and cortex. The activity of the root, which is a sink tissue, was high in all parts, but especially the highest in the root tip part. It is thought that this is because it helps unloading of sucrose in sink tissues that require sucrose degradation and plays a role in hydrolysising sucrose.

Morphological aspects of small intestinal mucosal injury and repair after electron irradiation

  • Grigory Demyashkin;Elza Karakaeva;Siuzanna Saakian;Vladimir Shchekin;Emir Elbuzdukaev;Umar Bamatgiraev;Daniia Ashgalieva;Makka Evsultanova;Daniil Kovalev;Darya Kabanova;Oleg Shatunov;Dmitrii Atiakshin
    • Anatomy and Cell Biology
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    • v.57 no.3
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    • pp.384-391
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    • 2024
  • Morphological evaluation of the small intestine mucosa and apoptosis activity (caspase-3) is necessary to assess the severity of damage to the small intestine. At the same time, proliferative index based on Ki-67 can be used to assess the regenerative potential of the small intestine. Fragments of small intestine of Wistar rats (n=60) of three groups: I) control (n=20); II) experimental group (n=20; local single electron irradiation at a dose of 2 Gy), III) experimental group (n=20; local single electron irradiation at a dose of 8 Gy) were studied by light microscopy using hematoxylin and eosin staining and immunohistochemical reactions with antibodies to Ki-67 and caspase-3. In all samples of the experimental groups, a decrease in all morphometric indices was observed on day 1 with a tendency to recover on day 3. Small intestinal electron irradiation led to disturbances in the histoarchitecture of varying severity, and an increase in cell apoptosis was observed (increased expression of caspase-3 and decrease in Ki-67). In addition, modulation of the PI3K/AKT and MAPK/ERK signaling pathways was detected. The most pronounced destructive changes were observed in the group of 8 Gy single electron irradiation. Local irradiation of the small intestine with electrons at a dose of 2 and 8 Gy results in a decrease in the number of enterocytes, mainly stem cells of the intestinal crypts.

Biological and Functional Characteristics of Lactic Acid Bacteria in Different Kimchi (김치 종류에 따른 유산균의 생물학적 및 기능적 특성)

  • Ko, Kang Hee;Liu, Wenli;Lee, Hyun Hee;Yin, Jie;Kim, In Cheol
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.1
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    • pp.89-95
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    • 2013
  • Biological and functional characteristics of lactic acid bacteria (LAB) were investigated in mustard stem/leaf kimchi (MK), cabbage kimchi (CK), young radish kimchi (YRK), and cubed radish kimchi (CRK). LAB of young radish kimchi were mainly composed of bacilli in contrast to the other kimchi. 89.2% LAB isolated from all kimchi harbored plasmids. However, LAB had an average of $4.1{\pm}0.5$ plasmid bands in YRK, more than MK, CK, and CRK. Exopolysaccharides were produced by 10.9~11.1% of LAB, and were especially by LAB isolated from radish kimchi. A significant percentage of LAB (69.5%) had antibacterial activity against one sensitive strain or more. LAB from CK, YRK and CRK had antimicrobial activities against Bacillus sp., Listeria monocytogenes, and Salmonella Typhimurium, while the LAB from MK had activities against Vibrio parahaemolyticus higher than those from the other kimchi. In YRK and CRK, acid-tolerant LAB were twice as prevalent as those in MK and CK. Bile-tolerant LAB isolated from CRK were more prevalent than other kimchi. When $10^8$ CFU of LAB were added to Caco-2 cells, 12.1% of LAB isolated from all kimchi showed similar adherent activity to Lactobacillus rhamnosus GG. LAB of MK particularly adhered to Caco-2 cells, 2.0~4.1 fold higher than LAB in the other kimchi. From these results, biological and functional characteristics of LAB varied according to the type of kimchi and LAB existing in kimchi were limited to their respective species.