Human umbilical cord is easy to obtain because it is discarded after birth, so that ethical issues can be avoided. Chondrogenesis studies using MSCs from bone marrow, cord blood, and adipose have indicated that TGFβ3 and BMP6 stimulate chondrogenesis. Therefore, we investigated chondrogenesis of hUC-MSCs on TGFβ3, BMP6, and combination of the two growth factors. We initiated chondrogenesis of cells by application of physical forces to form 3D cell clusters. After initiation, we designated four experimental groups for differentiation of cells, as follows: control, 10 ng/mL TGFβ3, 100 ng/mL BMP6, and the combination of 5 ng/mL TGFβ3 and 50 ng/mL BMP6. For analysis of chondrogenesis, GAG contents, mRNA expression, histological analysis and immunohistochemistry (IHC) were performed. For analysis of GAG contents, GAG assay was performed and RT-PCR was performed for determination of chondrogenic markers. Histological analysis was performed through safranin O, alcian blue, and IHC was performed using collagen type I and II. GAG contents were increased 184% by TGFβ3, 147% by BMP6, and 189% by the combination of TGFβ3 and BMP6, compared to control. The growth factors improved collagen II and aggrecan expression; in particular, TGFβ3 and BMP6 showed a synergistic effect, compared to only TGFβ3 or BMP6 treated. The results of histological and IHC analysis indicated that chondrogenic differentiation in TGFβ3 and the combination of TGFβ3 and BMP6 showed more cartilage deposition. In conclusion, TGFβ3 and BMP6 differentiated hUC-MSCs into chondrogenic clusters of the combination treatment of the two growth factors showed more efficient chondrogenic ability.

We examined the effects of biological activity Phellinus linteus mycelium culture with cassiae semen extract. Firstly, the optimal temperature, initial pH and culture period for mycelial growth in a liquid culture of P. linteus were determined, and they were 30℃, pH 5.0 and 8 days respectively. The five herbal materials were examined against several health functional efficacies, and, as a result, Cassiae semen was chosen, with its superior inhibitory effects in β-glucuronidase inhibitory activity, electron donating activity, ACE inhibitory, and α-glucosidase inhibitory activities(95.3%, 80.9%, 96.1 and 24.2%, respectively). P. linteus fruit body was investigated on β-glucuronidase inhibitory activity, electron donating activity, ACE inhibitory, and α-glucosidase inhibitory activities, and they were 54.7%, 81.9%, 30.0% and 20.1%, respectively. Accordingly, C. semen was used in the following experiment, to give an additive functional effect on the P. linteus. As the amount of C. semen in the cultural media increased, mycelial weight and β-glucan contents also increased, but final pH was not influenced. In addition, the β-glucuronidase inhibitory activity, electron donating activity, and α- glucosidase inhibitory activity increased. P. linteus mycelium culture showed higher activities in the other three tests above, except for electron donating activity, when C. semen was added to the medium before cultivation.

본 연구에서는 CCl4 존재하에서 Cl2(g) and ultra violet(200~300nm) 파장으로 maleic anhydride 와 Cl2(g)을 반응시켜 α,β-dichlorosuccinic acid 를 합성하였다. 그리고 두 번째 반응은 N-(α,β-dichloro)succinic acid 함유 glucosamine 유도체(I)를 합성하는 것으로 methanol 함유 2% acetic acid 용매 하에서 glucosamine과 과량의 α,β-dichlorosuccinic anhydride을 넣은 후 온도를 70℃로 올리면서 반응시켰다. 우리는 합성한 유기산 유도체들이 해태 김 재배와 피부미용에 주로 유용하게 활용할 것으로 본다.

Proteinuric conditions demonstrate structural and compositional changes of the foot processes and slit diaphragms between podocytes. β-Catenin in podocytes serves as an adapter protein anchoring P-cadherin of slit diaphragms to actin filaments of the podocyte cytoskeleton. To investigate the effect of ginseng total saponin (GTS) on pathologic changes of podocyte P-cadherin/β-catenin unit induced by diabetic conditions, we cultured mouse podocytes under: 1) normal glucose (5 mM, = control); 2) high glucose (HG, 30 mM); 3) advanced glycosylation end products (AGE)-added; or 4) HG plus AGE-added conditions and treated with GTS. In confocal imaging, β-catenin colocalized with P-cadherin predominantly at intercellular junction area. However, diabetic conditions relocalized and concentrated both molecules at perinuclear cytoplasmic area. In Western blotting, diabetic conditions, especially HG plus AGE-added condition, also decreased cellular β-catenin protein levels at 6, 24, and 48 hours. GTS improved such quantitative and qualitative changes of β-catenin. These findings imply that HG plus AGE have an influence on the redistribution and amount of P-cadherin/ β-catenin unit of podocytes, which can be reversed by GTS.

This study aims to examine the effect of Genetically Modified β -Carotene Biofortified Rice rice developed by simultaneous expression technology in NAAS on biological immunity. Accordingly, this study added Genetically Modified β-Carotene Biofortified Rice 25, 50% and general rice 50% as control group into diet and provided rats with the prescribed feeds and then measured the contents of immunoglobulin and cytokine in blood. As a result, male and female IgM, IgE, male IgG1, female IgG2a and TNF-a, IL5 and IL12 showed no significant difference; male IgG2a tended to decrease dependently on the combined concentration of Genetically Modified β-Carotene Biofortified Rice; female IgG1 showed significance with control group, but its association with diet was not found. The higher the dietary mixing ratio, the more the male and female IFN-a and female IL-4 contents, regardless of rice variety, and it was found that female IL6 content decreased significantly, but its association with diet was not found. The risk of beta carotene-enriched rice into environment and human body has not been reported yet. The digestion of Genetically Modified β-Carotene Biofortified Rice can be seen as "safe" as this test result showed no big difference between general rice and Genetically Modified β-Carotene Biofortified Rice, and its usability is full of suggestions.

Porcine blastocyst’s quality derived from in vitro is inferior to in vivo derived blastocysts. In this study, to improve in vitro derived blastocyst’s quality and then establish porcine ESCs (pESCs), we treated in vitro fertilized (IVF) embryos and parthenogenetic activated (PA) embryos with three chemicals: porcine granulocyte-macrophage colony stimulating factor (pGM-CSF), resveratrol (RES) and β-mercaptoethanol (β-ME). The control group was produced using M199 media in in vitro maturation (IVM) and porcine zygote medium-3 (PZM3) in in vitro culture (IVC). The treatment group is produced using M199 with 2 μM RES in IVM and PZM5 with 10 ng/mL pGM-CSF, 2 μM RES and 10 μM β-ME in IVC. Data were analyzed with SPSS 17.0 using Duncan’s multiple range test. In total, 1210 embryos in PA and 612 embryos in IVF evaluated. As results, we observed overall blastocyst quality was increased. The blastocyst formation rates were significantly higher (p<0.05) in the treatment groups (54.5%) compared to the control group (43.4%) in PA and hatched blastocysts rates in day 6 and 7 were also increased significantly. Total cell numbers of blastocyst were significantly higher (p<0.05) in the treatment group (55.1) compared to the control group (45.6). In IVF, hatched blastocysts rates in day 7 were increased significantly, too. After seeding porcine blastocyst, the attachment rates were higher in the treatment group (36.2% in IVF and 32.2% in PA) than the control group (26.6% in IVF and 19.5% in PA). Also, colonization rates and cell line derivation rates were higher in treatment group than control group. Colonization rates of control group were 10.8% in IVF and 2.4% in PA, but treatment group were 17.75% in IVF, and 13.1% in PA. And we investigated the correlation between state of blastocysts and attachment rate. The highest attachment rate is in hatched blastocyst (78.35±15.74 %). So, the novel system increased quality of porcine blastocysts produced from in vitro, subsequently increased attachment rates. The cell line derivation rates were 4.2% (IVF) and 2.4% (PA) in control group. In treatment group, they were 10.0% (IVF) and 7.2% (PA). We established 3 cell lines from PA blastocysts (1 cell line in control group and 2 cell lines in treatment group). All cell line has alkaline phosphatase activity and express pluri-potent markers. In conclusion, the novel system of IVM and IVC (the treatment of RES during IVM and RES, β-ME, and pGM-CSF during IVC) increased quality of porcine blastocysts produced from in vitro, subsequently increased derivation rates of porcine putative ESCs.

돼지는 인간과 생리적으로 유사하기 때문에 다양한 목적으로 연구되고 있다. 최근에는 돼 지를 이용한 이종장기이식 관련 연구가 큰 주목 받고 있으며, 치료용 단백질을 생산하기 위 한 생체반응기로써 이용되고 있다. 이러한 연구에 있어서 당 사슬의 역할은 매우 중요하다. 당 사슬은 치료용 단백질의 체내 안정성에 큰 영향을 미치며 다양한 방법으로 면역을 조절 한다고 알려져 있다. 하지만 돼지에서의 당 사슬 관련 연구는 미비하며, 많은 당 전이효소 들의 서열이나 기능이 정확히 분석되지 않고 있다. 따라서 이번 연구에서는 당 전이효소 중 하나인 β‐1,3‐N‐acetylglucosaminyltransferase 1 (B3GNT1)을 돼지로부터 동정 후 PK‐15 세 포주를 이용하여 기능분석을 하였다. 이 유전자는 다양한 glycan epitope를 형성하는데 있 어서 중요한 기능을 한다. 먼저 간 조직으로부터 획득된 cDNA를 주형으로 degenerated PCR을 수행하여 유전자를 동정하였다. 동정된 유전자는 368개의 아미노산을 encoding하 는 1227개의 nucleotide로 구성되어 있으며, 다른 종에서 보고된 B3GNT1과 높은 상동성을 가 지고 있는 것을 확인하였다. 또한, 기능 분석을 위하여 돼지 신장세포인 PK‐15에서 B3- GNT1의 과 발현을 유도하였다. RT‐PCR과 Western blot을 통하여 유전자의 과발현을 확인 하고, Lycopersicon esculentum lectin (LEA)를 이용한 ELISA 분석 방법을 통해 효소의 기 능을 확인하였다. B3GNT1은 poly N‐acetylactosamine (polyLacNAc) 형성에 중요한 역할 을 하기 때문에 이를 특이적으로 인지하는 LEA를 사용하였다. 이를 통해 B3GNT1의 과발현이 유도된 세포에서 더 많은 polyLacNAc이 합성되었음을 확인할 수 있었다. 또한, Gal(β1‐ 3)GalNAc structure를 이용한 기질반응을 통해 효소의 기능을 확인하였다. 과발현이 유도 된 세포에서 약 3배 이상의 높은 기질 반응성을 보였으며, 이를 통해 돼지로부터 클로닝 된 B3GNT1의 기능을 확인할 수 있었다. 돼지로부터 당 전이효소를 동정하고 분석하는 연구는 생체반응기로써 돼지를 이용하는 다른 연구에 중요한 기반이 될 것이라고 생각한다

In the present study, a phylogenetic analysis was undertaken based on the internal transcribed spacer (ITS) rDNA and partial β-tubulin gene sequence of the Ganoderma species. The size of the ITS rDNA regions from different Ganoderma species varied from 625 to 673 bp, and those of the partial β-tubulin gene sequence were 419 bp. Based on the results, a phylogenetic tree was prepared which revealed that Korean Ganoderma lucidum strains belong in a single group along with a G. lucidum strain from Bangladesh.

Opioid receptors have been pharmacologically classified as µ, δ, κ and ε. We have recently reported that the antinociceptive effect of morphine (a µ-opioid receptor agonist), but not that of β-endorphin (a novel µ/ε-opioid receptor agonist), is attenuated by whole body irradiation (WBI). It is unclear at present whether WBI has differential effects on the antinociceptive effects of µ-, δ-, κ- and ε-opioid receptor agonists. In our current experiments, male ICR mice were exposed to WBI (5Gy) from a 60 Co gamma-source and the antinociceptive effects of opioid receptor agonists were assessed two hours later using the hot water (52℃) tail-immersion test. Morphine and D-Ala2,N-Me-Phe4,Gly-ol-enkephalin (DAMGO), [D-Pen2-D-Pen5]enkephalin (DPDPE), trans-3,4-Dichloro-N-methyl-N-[2-(1-pyrrolidinyl)- cyclohexyl]¬benzeneacetamide (U50,488H), and β-endorphin were tested as agonists for µ, δ, κ, and ε-opioid receptors, respectively. WBI significantly attenuated the antinociceptive effects of morphine and DAMGO, but increased those of β-endorphin. The antinociceptive effects of DPDPE and U50,488H were not affected by WBI. In addition, to more preciously understand the differential effects of WBI on µ- and ε¬opioid receptor agonists, we assessed pretreatment effects of β-funaltrexamine (β-FNA, a µ-opioid receptor antagonist) or β-endorphin1-27 (β-EP1-27, an ε-opioid receptor antagonist), and found that pretreatment with β-FNA significantly attenuated the antinociceptive effects of morphine and β endorphin by WBI. significantly reversed the β-EP1-27 attenuation of morphine by WBI and significantly attenuated the increased effects of β-endorphin by WBI. The results demonstrate differential sensitivities of opioid receptors to WBI, especially for µ- and ε-opioid receptors.

The objective of the present study was to evaluate the potential antidiabetic and antioxidant effect of the ethanol extract from Chrysanthemum cornarium L. var. spatiosum(CSE) against alloxan-induced oxidative stress in pancreatic β-cells, HIT-T15. In this study, the antidiabetic effect of CSE was examined using the 3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazoliu bromide(MTT) cell proliferation assay, lactate dehydrogenase(LDH) release assay, NAD+/NADH ratio and insulin secretion. To further investigate whether CSE is involved in the antioxidant activity of alloxan-damaged HIT-T15 cells, its antioxidant effect against alloxan-induced oxidative stress was measured in HIT-T15 cells by determining the levels of antioxidant enzymes including superoxide dismutase(SOD), glutathione S-transferase(GST), glutathione reductase(GR) and glutathione peroxidase(GPx). The results of this analysis showed that alloxan significantly decreased cell viability, increased LDH leakage, and lowered NAD+/NADH ratio and insulin secretion in HIT-T15 cells. However, CSE significantly increased the viability of alloxan-treated cells and lowered LDH leakage. The intracellular NAD+/NADH ratio and insulin secretion were also significantly increased by 1.7-fold and 1.3-fold, respectively, after treatment with 100 ㎍/㎖ CSE. The HIT-T15 cells treated with alloxan showed significant decreases in the activities of antioxidant enzymes, while CSE significantly elevated the levels of antioxidant enzymes. These findings suggest that CSE could have a protective effect against cytotoxicity and dysfunction of pancreatic cells in the presence of alloxan-induced oxidative stress.

Oral lichen planus (OLP) is an atypically keratinized and ulcerative lesion, producing severe pain and discomforts in the involved patients. Nevertheless, the etiological factor or the pathogenetic mechanism has not been clearly elucidated. In the present study, the different gene expressions were screened in 21 cases of OLP by immunohistochemical (IHC) array method using 80 antibodies, and found that the pathway of E-cadherin/β-catenin was abnormally expressed compared to the other essential genetic pathways. Particularly, the expressions of eIF5A, DHS, and DOHH, which are the biomarkers of protein translation, were remarkably reduced, nevertheless the expression of β-catenin was strongly positive in the 7 cases among 21 cases of OLP. The other expressions of p53, BCL-2, MDM-2, PAKT, BAX, BAK, BAD, NFkB, HO-1, etc, were usually weak or sparse, while the expressions of PCNA, CDK4, and HSP-70 were markedly increased. Taken together, it is presumed that the overexpression of β-catenin indicates the derangement of E-cad/β-catenin/NFkB pathway, causing the transcription of cellular proliferating genes in downstream events, i.e., PCNA and CDK4, and that it may be eventually relevant to the malignant potential of OLP epithelial cells. It is also suggested that the activation of β-catenin/TCF/LEF1 pathway be closely relevant to the immunological reaction of OLP with the accumulation of T-cells underneath the mucosal epithelium.

Tumor necrosis factor alpha (TNFα) is a multifunctional cytokine that is elevated in inflammatory diseases such as atherosclerosis, diabetes and rheumatoid arthritis. Recent evidence has suggested that β2 adrenergic receptor(β2AR) activation in osteoblasts suppresses osteogenic activity. In the present study, we explored whether TNFα modulates βAR expression in osteoblastic cells and whether this regulation is associated with the inhibition of osteoblast differentiation by TNFα. In the experiments, we used C2C12 cells, MC3T3- E1 cells and primary cultured mouse bone marrow stromal cells. Among the three subtypes of βAR, β2 and β3AR were found in our analysis to be upregulated by TNFα. Moreover, isoproterenol-induced cAMP production was observed to be significantly enhanced in TNFα-primed C2C12 cells, indicating that TNFα enhances β2AR signaling in osteoblasts. TNFα was further found in C2C12 cells to suppress bone morphogenetic protein 2-induced alkaline phosphatase (ALP) activity and the expression of osteogenic marker genes including Runx2, ALP and osteocalcin. Propranolol, a β2AR antagonist, attenuated this TNFα suppression of osteogenic differentiation. TNFα increased the expression of receptor activator of NF-κB ligand (RANKL), an essential osteoclastogenic factor, in C2C12 cells which was again blocked by propranolol. In summary, our data show that TNFα increases β2AR expression in osteoblasts and that a blockade of β2AR attenuates the suppression of osteogenic differentiation and stimulation of RANKL expression by TNFα. These findings imply that a crosstalk between TNFα and β2AR signaling pathways might occur in osteoblasts to modulate their function.

Recurrence-metastasis status of squamous cell carcinoma of tongue is a challenging oncologic problem. This study examined the expression of E-cadherin/β-catenin cell adhesion complex in squamous cell carcinoma of the tongue through an immunohistochemical study. Twenty samples from 15 patients with squamous cell carcinoma of the tongue, who were treated at the Department of Oral and Maxillofacial Surgery, consisted of primary or recurrent tumors along with matched metastatic lymph nodes were retrieved for immunohistochemical staining and grouped based on recurrence-metastasis status.Differences in stain localization were noted in E-cadherin, β–catenin and phospho β–catenin staining between different tumor groups based on the recurrence-metastasis status. The number of phospho β-catenin stain positive cells was found to have a significant role in survival. E-cadherin confirms its role as a powerful individual differentiation indicator and the role of β-catenin specially the phospho type elicts interest