• Journal of Ginseng Research
• /
• 제44권3호
• /
• pp.475-482
• /
• 2020

Background: Active natural ingredients, especially small molecules, have recently received wide attention as modifiers used to treat neurodegenerative disease by promoting neurogenic regeneration of neural stem cell (NSC) in situ. 20(S)-protopanaxadiol (PPD), one of the bioactive ingredients in ginseng, possesses neuroprotective properties. However, the effect of PPD on NSC proliferation and differentiation and its mechanism of action are incompletely understood. Methods: In this study, we investigated the impact of PPD on NSC proliferation and neuronal lineage differentiation through activation of the Wnt/glycogen synthase kinase (GSK)-3β/β-catenin pathway. NSC migration and proliferation were investigated by neurosphere assay, Cell Counting Kit-8 assay, and EdU assay. NSC differentiation was analyzed by Western blot and immunofluorescence staining. Involvement of the Wnt/GSK3β/β-catenin pathway was examined by molecular simulation and Western blot and verified using gene transfection. Results: PPD significantly promoted neural migration and induced a significant increase in NSC proliferation in a time- and dose-dependent manner. Furthermore, a remarkable increase in anti-microtubule-associated protein 2 expression and decrease in nestin protein expression were induced by PPD. During the differentiation process, PPD targeted and stimulated the phosphorylation of GSK-3β at Ser9 and the active forms of β-catenin, resulting in activation of the Wnt/GSK-3β/β-catenin pathway. Transfection of NSCs with a constitutively active GSK-3β mutant at S9A significantly hampered the proliferation and neural differentiation mediated by PPD. Conclusion: PPD promotes NSC proliferation and neural differentiation in vitro via activation of the Wnt/GSK-3β/β-catenin pathway by targeting GSK-3β, potentially having great significance for the treatment of neurodegenerative diseases.

• Clinical and Experimental Reproductive Medicine
• /
• 제31권4호
• /
• pp.209-215
• /
• 2004

Objective: Embryonic stem (ES) cells could be differentiated into the specific cell types by alternation of culture condition and modification of gene expression. This study was performed to evaluate the differentiation protocol for mouse and human ES cells to insulin secreting cells. Methods: Undifferentiated mouse (JH-I) and human (Miz-hESI) ES cells were cultured on STO feeder layer, and embryoid bodies (EBs) were formed by suspension culture. For the differentiation, EBs were cultured by sequential system with three stage protocol. The differentiating ES cells were collected and marker gene expressions were analyzed by seIni-quantitative RT-PCR in each stage. Amount of secreted insulin levels in culture media of human ES cells were measured by human insulin specific RIA kit. Results: During the differentiation process of human ES cells, GATA-4, a-fetoprotein, glucose transporter-2 and Ngn-3 expression were increased whereas OctA was decreased progressively. Insulin and albuInin mRNAs were expressed from stage IT in mouse ES cells and from stage III in human ES cells. We detected 3.0~7.9 IlU/rnl secretion of insulin from differentiated human ES cells by in vitro culture for 36 days. Conclusion: The sequential culture system could induce the differentiation of mouse and human ES cells into insulin secreting cells. This is the fIrst report of differentiation of human ES cells into insulin secreting cells by in vitro culture with serum and insulin free medium.

• 한국식품영양과학회지
• /
• 제42권12호
• /
• pp.2082-2087
• /
• 2013

오가피의 주요 성분으로 알려져 있는 eleutheroside B와 E 그리고 식물의 세포벽을 구성하는 ${\beta}$-glucan 함량을 강원도(A. senticosus), 충남(A. senticosus), 제주도(A. koreanum) 산지별 오가피의 뿌리, 줄기, 잎, 열매 부위별로 각각 분석한 결과, 충남 오가피는 eleutheroside B가 검출되지 않는 것에 반해 특이적으로 높은 eleutheroside E를 함유하고 있었으며, 제주도 산지 오가피는 뿌리, 줄기, 잎, 열매모든 부위에서 eleutheroside B와 E가 함께 검출되는 것으로 확인되었다. 서로 다른 산지에서 재배되었지만 지역과 품종에 상관없이 줄기 부위에 가장 많은 eleutheroside B와 E가 함유되어 있었으며, ${\beta}$-glucan 또한 강원도(7.46%), 충남(3.41%), 제주도(5.05%) 산지 오가피 모두 줄기 부위에 가장 많았다. 오가피는 같은 품종이더라도 재배지역에 따라 eleutheroside B와 E 그리고 ${\beta}$-glucan 함량에 차이를 보여주었고 산지와 품종에 상관없이 줄기 부위에 가장 많이 함유되어 있었다. 따라서 본 연구결과를 기초로 오가피 소재 제품을 만들기 위해서는 제품의 목적에 따라 산지 및 부위의 선택이 필요할 것으로 사료되며, 산지에 따른 유효성분 차이에 있어서 탐라오가피는 제주도 특산식물로 오가피를 이용한 제품개발 시 지표성분을 표준화하는데 있어서 용이할 것으로 판단된다.

• Restorative Dentistry and Endodontics
• /
• 제24권1호
• /
• pp.49-54
• /
• 1999

Ko, Lim found some differences in the concentrations of bone resorptive cytokines, especially IL-$1{\alpha}$ and IL-$1{\beta}$ in periapical lesions and inflamed pulps. And they suppose that these differences may be due to the type of cells which produce each cytokine. The purpose of this study was to analyze the human odontogenic cysts & cystic fluid for their contents of IL-$1{\alpha}$, IL-$1{\beta}$ and TNF-$1{\alpha}$ and to compare the concentrations of each cytokine according to the cytokine producing cells. The cystic tissues used in this experiment, were obtained from periapical surgery or cyst enucleation surgery. Cystic fluid was obtained from root canal during routine endodontic therapy(n=5). Cystic tissues were subdivided into two groups, inflammatory radicular cyst group(n=15) and developmental odontogenic keratocyst group(n=3). Normal periapical tissues of extracted third molar(n=5) were also obtained to be used as control group. Each specimen was incubated in 0.5ml homogenizing buffer (0.1mol/L potassium chloride, 0.02mol/L TRIS;pH=7.6) for two hours and then homogenized with glass homogenizer. Each specimen was centrifuged in a microcentrifuge for 3 minutes, and supernatants were extracted. The concentrations of cytokines were measured with R&D ELISA kit. The data were analyzed by Mann-Whitney U test for the differences among the diseases and t test for the correlations among each cytokine. Following results were obtained ; 1. For IL-$1{\alpha}$ and IL-$1{\beta}$, all experimental groups showed significantly higher concentrations of each cytokine than the control group (p<0.05). 2. In radicular cysts, the concentrations of IL-$1{\alpha}$ were higher than IL-$1{\beta}$, but not stastically significant (p>0.05). In odontogenic keratocysts, the concentrations of IL-$1{\alpha}$ were significantly higher than IL-$1{\beta}$ (p<0.05). In cystic fluid, the concentration of IL-$1{\beta}$ was significantly higher than IL-$1{\alpha}$ (p<0.05). 3. Between odontogenic keratocysts and radicular cysts, the concentrations of IL-$1{\alpha}$ were significantly higher in odontogenic keratocysts than in radicular cysts (p<0.05). 4. For TNF-${\alpha}$, only cystic fluid group showed significantly higher concentrations than the control group (p<0.05).

• Restorative Dentistry and Endodontics
• /
• 제24권1호
• /
• pp.40-48
• /
• 1999

This study was designed to examine the specificities of the designed primers for F. nucleatum and F. necrophorum and to compare the PCR results using clinical samples with those of the anaerobic culture method. F. nucleatum and F. necrophorum spp. are frequently isolated in infected root canals, and they are related to periapical diseases. F. nucleatum(VPI 10197) and F. necrophorum(ATCC 25286) were used as references for PCR reaction, and thirty five teeth with one canal and periapical lesion were used. The samples were cultured anaerobically and identified using Rapid ID 32A(BioMerieux Vitek, Inc., France) as biochemical battery. In the GenBank database, species-specific PCR primers(nuc1/nuc2 primers for F. nucleatum and nec1/nec2 primers for F. necrophorum) were designed from the 16S ribosomal DNA(rDNA) sequences of F. nucleatum(accession number M58683) and F. necrophorum(accession number AF044948). PCR procedures of F. nucleatum(VPI 10197) and F. necrophorum (ATCC 25286) were simulated on a computer software. Amplify(v.1.2${\beta}$ for Macintosh). 820 bps and 817 bps of nucleotides were expected, respectively. Using extracted DNAs with QiaAmp tissue kit(Qiagen co., Germany), PCR was done. The results were as follows : 1. The nuc1/nuc2 primers produced an amplicon of 820 bps and the nec1/nec2 primers produced an amplicon of 817 bps. 2. The nuc1/nuc2 primers and the nec1/nec2 primers were specific and did not react with species other than the designated ones(i.e. nuc1/nuc2 primers did not produce amplicons for F. necrophorum, and vice versa.). And the PCR products of Porphyromonas endodontalis(ATCC 35406), Porphyromonas gingivalis(ATCC 33277), Prevotella intermedia(ATCC 25611), and Prevotella nigrescens(ATCC 33563), frequently isolated in infected root canals and periapical lesions, were not amplified by the primers specific for Fusobacterium nucleatum and Fusobacterium necrophorum. 3. This method utilizing PCR could detect F. nucleatum and F. necrophorum in clinical samples, while anaerobic culture method could detect neither.

• Biomolecules & Therapeutics
• /
• 제28권3호
• /
• pp.259-266
• /
• 2020

The present research work primarily investigated whether spinosin has the potential of improving the pathogenesis of Alzheimer's disease (AD) driven by β-amyloid (Aβ) overproduction through impacting the procession of amyloid precursor protein (APP). Wild type mouse Neuro-2a cells (N2a/WT) and N2a stably expressing human APP695 (N2a/APP695) cells were treated with spinosin for 24 h. The levels of APP protein and secreted enzymes closely related to APP procession were examined by western blot analysis. Oxidative stress related proteins, such as nuclear factor-erythroid 2-related factor 2 (Nrf2), and heme oxygenase-1 (HO-1) were detected by immunofluorescence assay and western blot analysis, respectively. The intracellular reactive oxygen species (ROS) level was analyzed by flow cytometry, the levels of Aβ_1-42 were determined by ELISA kit, and Thioflavin T (ThT) assay was used to detect the effect of spinosin on Aβ_1-42 aggregation. The results showed that ROS induced the expression of ADAM10 and reduced the expression of BACE1, while spinosin inhibited ROS production by activating Nrf2 and up-regulating the expression of HO-1. Additionally, spinosin reduced Aβ_1-42 production by impacting the procession of APP. In addition, spinosin inhibited the aggregation of Aβ_1-42. In conclusion, spinosin reduced Aβ_1-42 production by activating the Nrf2/HO-1 pathway in N2a/WT and N2a/APP695 cells. Therefore, spinosin is expected to be a promising treatment of AD.

• 대한약리학회지
• /
• 제31권3호
• /
• pp.285-290
• /
• 1995

이 실험은 여러 파장$(240{\sim}520\;nm)$의 자외선 또는 가시광선(이후 '광선'이라 표기함)을 흰쥐흉부대동맥에 조사하여 이때의 혈관장력의 변화 및 조직내 cyclic GMP농도의 변화를 관찰하기위하여 시행하였다. 돼지관상동맥 또는 흰쥐 흉부대동맥의 환상표본에 spectrofluorometer의 xenon lamp를 이용하여 여러 파장의 광선을 조사하고 이때의 장력변동을 polygraph상에 기록하였다. Cyclic GMP농도변화는 표본에 광선을 조사한 직후 조직을 얼리고 homogenization 및 원침시킨 후 상청액을 ether로 추출하여 RIA kit로 측정하였다. Phenylephrine으로 수축된 내피존재 흰쥐 흉부 대동맥에서는 광선조사로 수축반응을 보였고 320 nm에서 최대수축반응을 일으켰다. 그 이상의 파장에서는 점차 수축반응이 감소되어 420 nm에서는 최대 이완반응을 일으킨 후 점차 기본장력으로 회복되었다. 그러나 내피제거 표본에서는 전파장에서 이완반응만을 일으켰고 이때 최대 이완반응은 370 nm에서 관찰되었다. 내피존재 표본에서 320, 380 및 420 nm의 광선을 30초간 조사한 결과 380과 420nm에서 현저한 cyclic GMP의 증가가 관찰되었으나 320 nm에서는 유의한 변동이 없었다. 한편, 내피제거 표본에서는370 nm의 광선조사로 cyclic GMP함량이 약 4배 증가하였다. 이상의 성적으로부터 흰쥐 흉부대동맥은 광선조사에 의하여 내피존재 표본에서는 수축-이완의 이상성반응을, 제거표본에서는 이완반응만을 일으키고 양 표본의 이완반응은 nitric oxide-cyclic GMP계의 활성화에 기인하나 수축반응은 cyclic GMP계와 직접 관련성이 없는 것으로 추론하였다.

• 대한임상검사과학회지
• /
• 제39권3호
• /
• pp.210-216
• /
• 2007

There is increasing evidence that stromal reaction in cancer has an important diagnostic and prognostic significance. The aim of our study is to analyze the relation between the increase in mast cell number and the expression CD34 and alpha-smooth muscle actin (${\alpha}$-SMA) in the stroma of cervical intraepithelial neoplasia (CIN) and squamous cell carcinoma (SCC). We investigated a total of 29 CIN (1,2,3) and 21 SCC (microinvasive and invasive) specimens and compared the distribution of $CD34^+$ stromal cells, ${\alpha}-SMA^+$ cells, transforming growth factor-${\beta}1$ $(TGF-{\beta}1)^+$ cells, and the density of mast cells using immunohistochemistry with antibodies against CD34, ${\alpha}$-SMA, TGF-${\beta}1$, and c-Kit (CD117) respectively. Computerized image analysis was to evaluate the positive area (%) and density of the respective immunoreactive cells. In CIN $CD34^+$ cells were abundant in the stroma but no ${\alpha}-SMA^+$ cells were identified except the wall of blood vessels. $CD34^+$ cells were progressively decreased along the continuum from CIN 2 to microinvasive SCC and not observed in the stroma of invasive SCC. Whereas ${\alpha}-SMA^+$ cells were only observed in the stroma of microinvasive and invasive SCC. We found more intense TGF-${\beta}1$ expression in the increased mast cells in the stroma of invasive SCCs than that in the stroma of CIN. These results indicate that disappearance of $CD34^+$ stromal cells and appearance of ${\alpha}-SMA^+$ cells are associated with the stromal change of CIN to SCC and the transformation of $CD34^+$ stromal cells into ${\alpha}-SMA^+$ cells is mediated by TGF-${\beta}1$ secretions in the stromal mast cell of SCC.

• 대한치과보철학회지
• /
• 제45권4호
• /
• pp.419-430
• /
• 2007

Purpose: This in vitro study evaluated shear bond strengths of surface treatment porcelains with four porcelain repair systems simulating intraoral bonding of composite resin to feldspathic porcelain or pressable porcelain. Material and methods: Eighty Porcelain disks were prepared. Group A: forty disk specimens were fabricated with Feldspathic Porcelain($Omega^{(R)}900$, Vident, Menlo Park, CA, USA). Group B: forty disk specimens were fabricated with Pressable Porcelain(IPS Empress 2 ingot, Ivoclar-Vivadent, Schaan, Liechtenstein, Germany). Each groups was divided into 4 subgroups and composite resin cylinders were bonded to specimen with one of the following four systems: Clearfil Porcelain Bond(L. Morita, Tustin, CA, USA), Ulradent Porcelain Etch. (Ultradent, Salt Lake City UT, USA), Porcelain Liner-M(Sun Medical Co., Kyoto, Japan), Cimara Kit(Voco, Germany). After surface conditioning with one of the four porcelain repair systems substrate surfaces of the specimen were examined microscopically(SEM). Shear bond strengths of specimens for each subgroup were determined with a universal testing machine (5mm/min crosshead speed) after storing them in distilled water at $37{\pm}1^{\circ}C$ for 24 hours. Stress at failure was measured in $MP_a$, and mode of failure was recorded. Differences among four repair systems were analyzed with two way ANOVA and Duncan test at the 95% significance level. Results: In the scanning electron photomicrograph of the treated porcelain surface, hydrofluoric acid etched group appeared the highest roughness. The shear bond strength of the phosphoric acid etched group was not significantly(p>0.05) different between feldspathic porcelain and pressable porcelain. But in no treatment and roughened with a bur group, the shear bond strength of the feldspathic porcelain was significantly higher than that of the pressable porcelain. In hydrofluoric acid etched group, the shear bond strength of the pressable porcelain was significantly higher(p<0.05). Conclusion: 1. Treatment groups showed significantly greater shear bond strengths than no treatment group(p<0.05). 2. Group with more roughened porcelain surface did not always show higher shear bond strengths. 3. In phosphoric acid etched group, there was no significant difference in shear bond strength between feldspathic porcelain and pressable porcelain(p>0.05). However in the other groups, there were significant differences in shear bond strengths between feldspathic porcelain and pressable porcelain(p<0.05).

• 생명과학회지
• /
• 제15권1호
• /
• pp.152-159
• /
• 2005

본 연구에서는 생명과학분야의 연구활동 활성화에 기여코자 생명과학 분야의 연구비 지원 추이와 현황을 파악하여 생명과학분야 연구활동 활성화를 위한 방향을 제시하고자 과학재단 연구비 수혜자를 중심으로 자료를 분석하였다. 생명과학 분야에서는 개인단위보다는 집단 및 그룹단위 형태의 연구에 더 많은 연구비가 투자되고 있는 것으로 조사되었으며, 생명과학 분야의 단위과제당 연구비 수준은 전체 이공계의 연구비 수준에 비교해서 상대적으로 약간 낮은 수준$(2003년,\;88.0\%)$이었다. 각 세부분야별로 연구비지원액 및 지원과 제수를 기준으로 살펴볼 때, 기초의약학 분야의 약진이 두드러진 반면에 농수산 및 생물분야는 감소추세인 것으로 나타났다. 생명과학 분야의 연구활동 활성화를 위해서는 중 $\cdot$장기 연구전략계획 수립, 생명과학전문인력 DB구축과 타 분야 연구진 또는 생명과학 세부분야간의 연계 활용, 전략적 연구지원 분야의 도출 및 절정 연구지원단가 산출, 생명과학 기초연구 특별프로그램 개발, 학회 내 정책기획 분과 신설, 평가문화의 개선 및 생명과학 연구활동의 계량적 성과 지표 개발 등에 따르는 본 분야의 연구지원을 위한 시스템의 구축과 활용이 필요하다.