• Journal of Nutrition and Health
• /
• v.47 no.3
• /
• pp.167-175
• /
• 2014

Purpose: Previous studies have shown that treatment with Smilax china L. leaf extract (SCLE) produces antidiabetic effects due to ${\alpha}$-glucosidase inhibition. In this study, we examined the mechanism underlying these antidiabetic effects by examining glucose uptake in HepG2 cells cultured with SCLE. Methods: Glucose uptake and glucokinase activity were examined using an assay kit. Expression of glucose transporter (GLUT)-2, GLUT-4, and HNF-$1{\alpha}$ was measured by RT-PCR or western blot. Results: Treatment with SCLE resulted in enhanced glucose uptake in HepG2 cells, and this effect was especially pronounced when cells were cultured in an insulin-free medium. SCLE induced an increase in expression of GLUT-2 but not GLUT-4. The increase in the levels of HNF-$1{\alpha}$, a GLUT-2 transcription factor, in total protein extract and nuclear fraction suggest that the effects of SCLE may occur at the level of GLUT-2 transcription. In addition, by measuring the change in glucokinase activity following SCLE treatment, we confirmed that SCLE stimulates glucose utilization by direct activation of this enzyme. Conclusion: These results demonstrate that the potential antidiabetic activity of SCLE is due at least in part to stimulation of glucose uptake and an increase in glucokinase activity, and that SCLE-stimulated glucose uptake is mediated through enhancement of GLUT-2 expression by inducing expression of its transcription factor, HNF-$1{\alpha}$.

• Journal of dental hygiene science
• /
• v.15 no.2
• /
• pp.209-219
• /
• 2015

In order to explore an effect of interaction of Streptococcus gordonii, Fusobacterium nucleatum and Porphyromonas gingivalis that are bacteria relevant to periodontal disease on its growth, the bacteria were incubated in trypticase soy hemin menadione broth at $37^{\circ}C$ $CO_2$ incubator for 7 days through anaerobic jar by single and co-culture with heat treated dead bacteria under anaerobic gas pack. In order to confirm growth level, absorbance was measured and for confirming colony structure and form, it was observed with scanning electron microscope. In order to confirm an effect on pathogenicity of P. gingivalis, real time reverse transcriptase polymerase chain reaction was implemented for expression analysis for rgpA gene that produces HRgpA which is gingipain. As a result, the following conclusion was obtained. Colony formation of S. gordonii and P. gingivalis was increased by other dead bacteria and in case of F. nucleatum, its colony formation was showed an aspect of being increased by dead bacterium of P. gingivalis but decreased by dead bacterium of S. gordonii. Therefore, it is considered that the strains being used for this study would affect interactively through bacterial cell itself as well as their interaction factor at the time of colony formation.

• Polymer(Korea)
• /
• v.35 no.6
• /
• pp.499-504
• /
• 2011

Articular cartilage has an intrinsic difficulty in recovering damages, which requires its tissue engineering treatment. Demineralized bone particle (DBP) contains various bioactive molecules. It is widely used biomaterials in the field of tissue engineering. We developed the synthetic/natural hybrid scaffolds with poly(lactide-co-glycolide) (PLGA) and solution of DBP. The chondrocytes were seeded on the PLGA-DBP scaffolds and MTT assay, morphological observation, biological assay for collagen, sGAG, and RT-PCR were performed to analyze the effect of the DBP on cell viability and extracellular matrix secretion. In SEM observation, we observed that PLGA-DBP scaffolds had uniform porosity. As MTT assay showed scaffolds containing DB solution had higher cell viability then only PLGA scaffolds. The PLGA-DBP scaffolds had better ECM production than PLGA scaffold. It was proven by the higher specific mRNA expression in the PLGA-DBP scaffold than that in PLGA scaffold. These results indicated that PLGA-DBP scaffolds might serve as potential cell delivery vehicles and structural bases for in vitro tissue engineered articular cartilage.

• Experimental and Molecular Medicine
• /
• v.50 no.12
• /
• pp.13.1-13.12
• /
• 2018

We aimed to explore whether changes in circulating levels of miRNAs according to type 2 diabetes mellitus (T2DM) or prediabetes status could be used as biomarkers to evaluate the risk of developing the disease. The study included 462 patients without T2DM at baseline from the CORDIOPREV trial. After a median follow-up of 60 months, 107 of the subjects developed T2DM, 30 developed prediabetes, 223 maintained prediabetes and 78 remained disease-free. Plasma levels of four miRNAs related to insulin signaling and beta-cell function were measured by RT-PCR. We analyzed the relationship between miRNAs levels and insulin signaling and release indexes at baseline and after the follow-up period. The risk of developing disease based on tertiles (T1-T2-T3) of baseline miRNAs levels was evaluated by COX analysis. Thus, we observed higher miR-150 and miR-30a-5p and lower miR-15a and miR-375 baseline levels in subjects with T2DM than in disease-free subjects. Patients with high miR-150 and miR-30a-5p baseline levels had lower disposition index (p = 0.047 and p = 0.007, respectively). The higher risk of disease was associated with high levels (T3) of miR-150 and miR-30a-5p ($HR_{T3-T1}=4.218$ and $HR_{T3-T1}=2.527$, respectively) and low levels (T1) of miR-15a and miR-375 ($HR_{T1-T3}=3.269$ and $HR_{T1-T3}=1.604$, respectively). In conclusion, our study showed that deregulated plasma levels of miR-150, miR-30a-5p, miR-15a, and miR-375 were observed years before the onset of T2DM and pre-DM and could be used to evaluate the risk of developing the disease, which may improve prediction and prevention among individuals at high risk for T2DM.

• Journal of Chitin and Chitosan
• /
• v.23 no.4
• /
• pp.262-266
• /
• 2018

Even though the saps have been consumed as beverage in Korea, however a little research has been conducted on the chemical composition. We determined free sugars, mineral in the sap from Juglans mandshurica Maxim. collected from BongHwa region. The contents of glucose, sucrose, and fructose in the sap from J. mandshurica were 0.15, 2.73, and 0.09%, respectively. The prominent minerals in the sap from J. mandshurica were calcium and potassium. The contents of Ca, K, Mg, Fe, and Mn of sap from J. mandshurica were 0.61, 0.57, 0.12, 0.002, and 0.014 ppm, respectively. The sap from J. mandshurica show inhibitory effect on elastase. And real-time RT-PCR showed that sap from J. mandshurica increased mRNA level of AQP3 and HAS2 gene and increased hyaluronic acid production in HaCaT cells. These results indicated that the sap from J. mandshurica can potentially be used for developing cosmetic ingredient for skin moisturizing and anti-aging.

• Development and Reproduction
• /
• v.22 no.4
• /
• pp.379-391
• /
• 2018

Through the development of organic synthetic skill, chemicals that mimic signaling mediators such as steroid hormones have been exposed to the environment. Recently, it has become apparent that this circumstance should be further studied in the field of physiology. Estrogenic action of chronic low-dose nonylphenol (NP) and di-(2-ethylhexyl) phthalate (DEHP) in mouse uterus was assessed in this study. Ten to twelve-week-old female mice (CD-1) were fed drinking water containing NP (50 or $500{\mu}g/L$) or DEHP (133 or $1,330{\mu}g/L$) for 10 weeks. Uterine diameter, the thickness of myometrium and endometrium, and the height of luminal epithelial cells were measured and the number of glands were counted. The expression levels of the known $17{\beta}$-estradiol ($E_2$)-regulated genes were evaluated with real-time RT-PCR methodology. The ration of uterine weight to body weight increased in $133{\mu}g/L$ DEHP. Endometrial and myometrial thickness increased in 133 and $1,330{\mu}g/L$ DEHP treated groups, and in 50, $500{\mu}g/L$ NP and $133{\mu}g/L$ DEHP, respectively. The height of luminal epithelial cell decreased in NP groups. The numbers of luminal epithelial gland were decreased in NP groups but increased in $50{\mu}g/L$ DEHP group. The histological characters of glands were not different between groups. The mRNA expression profiles of the known $17{\beta}$-estradiol ($E_2$) downstream genes, Esr1, Esr2, Pgr, Lox, and Muc1, were also different between NP and DEHP groups. The expression levels dramatically increased in some genes by the NP or DEHP. Based on these results, it is suggested that the chronic low-dose NP or DEHP works as estrogen-like messengers in uterus with their own specific gene expression-regulation patterns.

• Clinical and Experimental Reproductive Medicine
• /
• v.32 no.2
• /
• pp.133-147
• /
• 2005

Objectives: This study was carried out to establish human embryonic stem cells derived from frozen-thawed embryos using mouse embryonic fibroblasts (mEFs), human fetal skin and muscle fibroblasts as feeder cells, and to identify the characteristic of embryonic stem cells. Methods: When primary mEFs, human fetal skin and muscle fibroblasts were prepared, passaging on 4 days from replating could have effective trypsinization and clear feeder layers. Eight of 23 frozenthawed 4~8 cell stage embryos donated from consenting couples developed to blastocysts. Inner cell mass (ICM) was isolated by immunosurgery. ICM was co-cultured on mEFs, human fetal skin or muscle fibroblasts. The ICM colonies grown on mEFs, human fetal skin or muscle fibroblasts were tested the expression of stage specific embryonic antigen-3, -4 (SSEA-3, -4), octamer binding transcription factor-4 mRNA (Oct-4) and alkaline phosphatase surface marker. Results: We obtained 1 ICM colony from 2 ICM co-cultured on mEFs as feeder cells and did not obtain any ICM colony from 6 ICM clumps co-cultured on human fetal skin or muscle fibroblasts. The colony formed on mEFs could be passaged 30 times every 5 days with sustaining undifferentiated colony appearance. When the colonies cultured on mEFs were grown on human fetal skin or muscle fibroblasts, the colonies could be passaged 15 times every 9 days with sustaining undifferentiated colony appearance. The colonies grown on mEFs and human fetal fibroblasts expressed SSEA-4 and alkaline phosphatase surface markers and positive for the expression of Oct-4 by reverse transcription-polymerase chain reaction (RT-PCR). The produced embryoid body differentiated spontaneously to neural progenitorlike cells, neuron-like cells and beating cardiomyocyte-like cells, and frozen-thawed embryonic stem cells displayed normal 46,XX karyotype. Conclusions: The human embryonic stem cells can be established by using mEFs and human fetal fibroblasts produced in laboratory as feeder cells.

• Clinical and Experimental Reproductive Medicine
• /
• v.32 no.2
• /
• pp.121-132
• /
• 2005

연구목적: 본 연구에서는 비폐쇄성 무정자증 환자에서 나타나는 정자형성과정의 이상과 고환세포의 세포자연사와의 연관관계 여부를 확인하였다. 또한 SELDI-TOF MS 분석을 통하여 고환 내 단백질 발현 양상을 확인하고, 질환에 따른 효과적인 biomarker 개발 가능성 여부를 확인하였다. 재료 및 방법: RT-PCR 및 면역조직화학법을 사용하여 고환에서의 Fas, FasL, Bcl-2, Bax와 Caspase-3의 발현 양상을 확인하고, in situ DNA 3'-end-labelling 방법으로 고환세포의 세포자연사 양상을 확인하였다. SELDI-TOF MS 분석법에 의한 고환의 병리학적 소견에 따른 단백질 발현 변화는 소수성 칩 ($H_4$)을 사용하여 분자량 10~100 kDa 범위 내에서 분석하였다. 결 과: 정상적인 정자형성과정을 보이는 폐쇄성 무정자증 환자의 고환에 비해 지주세포 증후군 (Sertoli cell only syndrome)과 성숙정지 (maturation arrest)를 보이는 고환 내 생식세포와 지주세포에서 세포자연사가 현저하게 증가한 것을 확인할 수 있었다. 세포자연사 관련인자들의 발현 양상을 확인한 결과, 지주세포 증후군과 성숙정지 환자군에서 Fas와 FasL mRNA의 발현이 증가하였으나, bcl-2, bax와 caspase-3 mRNA 발현의 경우에는 두 질환 모두에서 유의한 차이를 확인할 수 없었다. FasL 단백질 발현의 경우, 세포자연사의 증가가 관찰되었던 지주세포 증후군과 성숙정지를 보이는 환자의 간질세포와 지주세포에서 증가하는 양상을 나타내었다. SELDI-TOF MS 분석 결과에서 폐쇄성 무정자증 환자군에 비해 전체적인 단백질 발현양이 지주세포 증후군과 성숙정지 환자의 고환에서 감소하는 양상을 보였으며, 특히, 16.730 kDa 단백질의 현저한 감소를 확인할 수 있었다. 결 론: 본 연구결과를 통해 비폐쇄성 무정자증 환자에서 나타나는 정자형성과정의 장애는 생식세포의 비정상적인 세포자연사와 연관되어 있으며, 고환 내 Fas와 FasL의 비정상적인 발현이 주된 원인인 것을 확인할 수 있었다. 또한, SELDI-TOF MS 분석법을 통한 단백질 발현 양상의 연구는 무정자증 환자에서의 다양한 병리학적 소견을 쉽게 파악할 수 있는 biomarker 발굴뿐만 아니라 질환의 원인규명을 위한 연구에도 유용하게 이용될 수 있을 것으로 사료된다.

• The Korean Journal of Malacology
• /
• v.29 no.3
• /
• pp.171-179
• /
• 2013

Arginine kinase (ArK) is known to play an important role in most invertebrates the level of ATP by phosphorylation of phosphagens in cell and immuninty in living organisms. ArK has been identified in many kinds of organisms ranging from invertebrate to vertebrate. However, no ArK gene has been cloned and investigated from N. samarangae. This leads us to identify ArK cDNA (NsArK) from the expressed sequence tag (EST) sequencing of N. samarangae. Sequence analysis indicated that the coding region of 1,065 bp contains 355 amino acid residues. Molecular phylogenetic analysis shows that NsArK had very high similarities with mollusca and arthropoda. In an attempt to investigate a potential role of NsArK in the digestive gland of N. samarangae, expression patterns were analyzed. RT-PCR analsysis shows that NsArK mRNA is induced in the rane of 1.2 fold at 6 hr by laminarin when compared with the control. The immunnologial and physiological role of NsArK remains to be further investigated in N. samarangae.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
• /
• v.23 no.1
• /
• pp.94-110
• /
• 2010

Objective : This study was performed to evaluate the effect of immune reaction inductive substances such as phorbol-myristate-acetate(PMA), lipopolysaccharide(LPS), dermato-phagoides pteronyssus crude extract(DPE), dinitrochloro-benzene(DNCB) and Cryptotympana pustulata(CP), the Cryptotympana pustulata extracting substance at simultaneously on the translocation of nuclear factor-kappa B(NF-${\kappa}B$) towards to the nucleus and the mRNA expression patterns of various cytokine genes in Human acute monocytic leukemia cell line(THP-1 cells), monocytes of human. Experiment : To analyze cytokine genes expression patterns, the RT-PCR method was used, measuring tumor necrosis factor(TNF)-$\alpha$ that had been secreted during cell culture in the ELISA method. The morphological change in the cell observed during THP-1 cell culture was observed using a scanning electron microscope (SEM) and the quantitative distribution in the cell NF-${\kappa}B$ was analyzed through immunocytochemistry and a confocal microscopy. Result : CP showed different influences onto the mRNA expression patterns of cytokine genes with PMA, LPS. DPE and DNCB according to the types of immune inductive substances in the THP-1 cells. The expressions of inter-leukin(IL)-10, interferon(INF)-$\gamma$, TNF-$\alpha$ and monocyte chemoattractantant protein(MCP)-1 induced by PMA were suppressed by CP while the expression of transforming growth factor(TGF)-$\beta$ was promoted. Regarding the secretion pattern of TNF-$\alpha$ according to PMA processing, its secretion amount was increased by CP concurrent processing, in case of processing CP onto PMA and LPS, We discovered that the secretion amount of TNF-$\alpha$ was increased. Upon processing PMA and LPS on the THP-1 cell strain at the same time or either additionally processing CP thereon, the movement increase towards the nucleus from the NF-${\kappa}B$ cell cytoplasm, a transcription factor was able to be observed. Conclusion : In this study, Cryptotympana pustulata extracting substance was confirmed that it had an influence on expression patterns of cytokine genes according to the actions of a variety kinds of immune reaction inductive substances processed on the monocyte THP-1 cell of humans. Therefore, additional studies as for the immune adjusting function of Cryptotympana pustulata are considered to be able to offer important materials for curing immune abnormal diseases such as atopy dermatitis afterward.