The purpose of this study was to evaluate the antioxidant and anti-adipogenic activities of Ligularia stenocephala (L. stenocephala) extract. The contents of the total polyphenol of the extract was 55.950 mg GAE/g residue. Antioxidant activities of L. stenocephala were evaluated by free radical scavenging ability and a reducing power test. 2,2'azino-bis- (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and α-α-diphenyl-β-picrylhydrazyl (DPPH) free radical scavenging activities of the extract were approximately 90% and 70%, respectively. Reducing power of the extract was 258.833 mg TE/g residue. The anti-adipogenic activity of L. stenocephala extract was examined in 3T3-L1 cells. During adipocyte differentiation, the 3T3-L1 cells were treated both with and without the extract. L. stenocephala extract suppressed the lipid accumulation in a concentration-dependent manner in the 3T3-L1 cells. The L. stenocephala extract inhibited the expression of peroxisome proliferator activated receptor γ (PPARγ) and adipocyte protein 2 (aP2) proteins, compared with control adipocytes. These results indicate that L. stenocephala could be regarded as a potential source natural antioxidant and an anti-obesity agent.

Amyloid-β protein (Aβ) is known to increase free radical production in neuronal cells, leading to cell death by oxidative stress. The purpose of this study was to evaluate the protective effects of PineXol® on Aβ25-35 induced neuronal cell death. Rat pheochromocytoma (PC-12) cells were pre-treated with 100 μg/mL of PineXol® for 2 h. The cells were exposed to single dose of 30 μM Aβ25-35 for 24 h. Cell death was assessed by a cell count kit-8 (CCK-8) assay, lactate and dehydrogenase (LDH) release assay. An Apoptotic process was analyzed by a protein expression of the Bcl-2 family using western blotting. Cell viability increased in PC-12 cells treated with both Aβ25-35 and PineXol®, compared to the control group. PineXol® induced a decrease of the Bcl-2 protein expression (p<0.05), while Bax and Sod1 increased (p<0.05), indicating attenuation of Aβ25-35 induced apoptosis. These results suggest that PineXol® may be a good candidate for the prevention of Alzheimer’s disease(AD).

This study examined the antioxidant and neuronal cell protective effects of the water and methanol extracts of Eugenia caryophyllata Thunb. The total polyphenol content was significantly higher in the methanol extract than in the water extract. The DPPH radical scavenging activity in the water extract was similar to Vit. C at a concentration of 100~200 μg/mL. The ABTS radical scavenging activity in the water and methanol extract was similar to Vit. C at a concentration of 800~1,000 μg/mL. The superoxide dismutase (SOD)-like activity in the methanol extract was similar to Vit. C at a concentration of 800~1,000 μg/mL. The DPPH, ABTS radical scavenging and (SOD)-like activity increased with increasing extract concentration. In a cell viability using MTT, the water extract (50 and 100 ppm) and methanol extract (100 ppm) had a protective effect against H2O2-induced neurotoxicity.The result ssuggest that the extract of E. caryophyllata Thunb. has antioxidant activities and may be useful for treating neurodegenerative disorders.

This study examined the relationship between dinoflagellate cysts and vegetative cells, to a certain extent, by conducting a germination experiment on dinoflagellate cysts collected from a sediment trap and surface sediment. The germination experiment showed that 56.8%, 25 of the 44 species of dinoflagellate cysts seen in the sediment trap, germinated, which confirmed the relationship between cysts and vegetative cells. The germination experiment also found that Votadinium carvum showed different forms of vegetative cells in all three forms of cysts, which required an accurate identification of the species through a genetic analysis. Furthermore, the species known to be the cyst of Cochlodinium polykrikoides was determined to be Cochlodinium sp., and the cysts of C. polykrikoides did not appear.

Acinic cell carcinoma is a well-differentiated, low-grade tumor that accounts for 1-3% of salivary gland tumors. Among the variant of acinic cell carcinoma, papillary cystic variant of acinic cell carcinoma is much more rare and it is known to be difficult to diagnose and has a poor prognosis. In this paper, we report a case of 58 - year - old man diagnosed as papillary cystic variant of acinic cell carcinoma and the characteristics of the lesion by integrating the recent cases of papillary cystic variant of acinic cell carcinoma. This study emphasizes the need for long-term studies and further investigation of papillary cystic variants of acinic cell carcinoma.

5-fluorouracil (5-FU) is a pyrimidine analog which can work as antineoplastic antimetabolite by blocking thymidylate synthetase conversion of deoxyuridylic acid to thymidylic acid in DNA synthesis. This study is aimed to know the anticancer effect of 5-FU on the expressions of important signaling proteins in KB cells through immunoprecipitation high performance liquid chromatography (IP-HPLC). KB cells were treated with 5 μM 5-FU and cultured for 12, 24, 48, 72, and 96 hours, and followed by IP-HPLC analysis using 32 antisera. 5-FU suppressed the proliferation of KB cells by decreases in the expressions of proliferation-related proteins, Ki-67, PCNA, CDK4, and MPM2 to 82.6%, 92.4%, 95.2%, and 95.9%, respectively, but increases of antiproliferation-related proteins, p16 and p21 to 106.7% and 125.5%, respectively, during 96 hours of experiment. This proliferation reduction was also negatively regulated by cMyc/MAX/MAD network signaling. The cellular protection and survival were consistently arrested by 5-FU treatment in KB cells. The expressions of NFkB, MDR, p-mTOR, and TNFα were decreased to 95.1%, 92.8%, 93.4%, and 90.3% in 48-72 hours, respectively, while cellular stress was increased by upregulation of p38 to 111.3% in 48 hours. And the expressions of pAKT1/2/3, hTERT, and AMPK were also decreased to 93.3%, 97.4%, and 89.3% in 24-48 hours, respectively, while the cellular transformation might be undergone by upregulation of TGF-β1 to 117% until 96 hours. Particularly, 5-FU treatment greatly induced the cellular apoptosis in KB cells by increased expressions of PARP, cPARP, caspase 9, c-caspase 9, caspase 8, and caspase 3 in the lack of p53/BAX and FASL/FAS signaling. The expressions of PARP and c-PARP were increased maximum to 119.2% in 24 hours, and followed by increases of caspase 9, c-caspase 9, caspase 8, and caspase 3 to 111.2%, 125.9%, 108.6%, and 116.3% in 72-96 hours. Therefore, it is presumed that 5-FU induced cellular apoptosis in KB cells may be derived from the overexpression of PARP due to the increased DNA defect caused by 5-FU, which can lead to ATP depletion and subsequent cellular apoptosis.

In this study, the inhibitory effect of Paecilomyces tenuipes extract on PSA and angiogenesis-related factor expression levels were investigated in human prostate cancer cells, LNCaP. P. tenuipes extract significantly inhibited PSA expression in a dose-dependent manner. We also investigated the inhibitory effect of P. tenuipes extract on the expression of angiogenesis-related genes including VEGF, MMP-2, MMP-9, TIMP-1, and TIMP-2. P. tenuipes extract significantly down-regulated the expression of MMP-2 and MMP-9 in a dose-dependent manner. On the contrary, P. tenuipes increased the expression of TIMP-1 and TIMP-2. Our findings indicate that P. tenuipes exhibits an inhibitory effect on angiogenesis in human prostate cancer cells.

Bropirimine, a class of antineoplastic agents, is known as one of the potent immunomodulators and is currently under clinical development for the treatment of cancer. However, the effect of bropirimine on the cow remains unknown as a therapeutics agent. In this experiment, the effect of bropirimine in the peripheral blood mononuclear cells (PBMCs) stimulated by lipopolysaccharide (LPS) or concanavalin-A (Con-A) was examined. Jugular venous blood was collected from Korean Hanwoo calves and PBMCs were isolated. It was used to study the effect of bropirimine upon stimulation with LPS or Con-A for 72 hours. The expression pro-inflammatory cytokines like Tumor Necrosis Factor α (TNF-α) and Interferon γ (IFN-γ) were confirmed. Bropirimine significantly inhibited LPS- or Con-A-induced TNF-α and Con-A-induced IFN-γ in dose-dependent manner. Furthermore, Bropirimine inhibited TNF-α and Con-A mRNA expression at the transcription level. These results clearly indicated that bropirimine inhibited LPS or Con-A stimulated up-regulation of proinflammatory cytokines in a dose-dependent manner without conspicuous cytotoxicity. The bropirimine has potential to protect cow from LPS or Con-A induced endotoxin shock, possibly through inhibition of the production of proinflammatory cytokines. It suggesting that bropirimine may be a novel therapeutic agent for the prevention of inflammatory diseases. This result revealed specific features of the immune responses depending on the bropirimine compound and would help to knowledge of bovine immunity.

Characteristics of induced pluripotent stem (iPS) cells are consistent with those of embryonic stem (ES) cells. However, exogenous genes integrated by using retrovirus delivery systems cannot be completely removed from the cells. In a recent report, activation-induced cytosine deaminase (AID) and thymine DNA glycosylase (TDG) can induce pluripotency state in mouse differentiated cells through the process of DNA demethylation. Thus, we hypothesized that the two reprogramming factors may convert efficiently bovine differentiated cells into pluripotency state. So, genes of AID and TDG were integrated into pCMV6-AC-IRES-GFP-Puro expression vector, which was transfected into bovine differentiated cells. As results, the colonies derived from AID+TDG-induced bovine cells were formed on day 7 after culture. The number of AP positively colonies in AID+TDG-induced bovine cells was significantly higher than in AID-induced bovine cells (p<0.05). Additionally, expression of pluripotent genes (OCT-3/4, NANOG, SOX2) was slightly increased in AID+TDG-induced bovine cells, as compared to AID-induced bovine cells. Protein expressions of OCT-3/4, NANOG and SOX2 in AID+TDG-induced bovine cells were slightly increased rather than AID-induced bovine cells. Finally, DNA demethylation in the promoter regions of pluripotent markers in AID+TDG-induced bovine cells was increased than that of AID-induced bovine cells. In conclusion, pluripotent stem cells could be efficiently produced from bovine differentiated cells by using non-integrating delivery system with the reprogramming factors (AID and TDG).

소화관에서 주로 발생하는 반지세포암은 대부분은 위에서 발생하며, 담낭에서 원발성으로 발생하는 경우는 극히 드물어 지금까지 국내에서는 1예만 보고되고 있다. 본 증례는 58세의 남자가 5일 전부터 발생한 상복부 통증을 주소로 내원하여 복부 전산화단층촬영과 복부 자기공명 담췌관 촬영술에서 담낭 결석을 동반한 담낭염 및 담관 결석으로 진단되었다. 내시경 역행 췌담관 시술을 시행하여 담관석을 제거한 후 복강경하 담낭절제술을 시행하였다. 절제된 담낭의 조직검사에서 반지 세포암으로 진단되어, 항암 치료 및 방사선 치료를 시행받았으나, 1년 후 전이성 병변 및 복수가 진행되어 현재 보존적 치료 중이다. 이에 저자들은 담낭염으로 오인된 담낭의 원발성 반지세포암 1예를 경험하여 문헌고찰과 함께 보고한다.

(-)-Epigallocatechin-3-gallate (EGCG) is a major catechin found in green tea. It is reported that EGCG possesses various health benefits including anti-cancer, antioxidant, anti-diabetes, and anti-obesity. The objective of this study was to investigate the effects of EGCG on adipogenesis via activation of AMP-activated protein kinase (AMPK) pathway in 3T3-L1 preadipocytes. In order to determine the effects of EGCG on adipogenesis, preadipocyte differentiation was induced in the presence or absence of EGCG (0~100 μM) for a period of 6 days. EGCG significantly inhibited fat accumulation and suppressed the expression of adipogenic specific proteins including peroxisome proliferator-activated receptor (PPAR)-γ. Also, EGCG markedly increased the activation of AMPK and acetyl-CoA carboxylase (ACC) and the production of intracellular reactive oxygen species (ROS). However, any pretreatment with a specific AMPK inhibitor, compound C, abolished the inhibitory effects of the EGCG on PPARγ expression. This study suggests that EGCG has anti-adipogenic effects through modulation of the AMPK signaling pathway and therefore, may be a promising antiobesity agent.

To compare functional Chinese cabbage(‘Amtak’ baechu; F1 hybrid cultivar between Brassica rapa and B. perkinensis, AB) with general Chinese cabbage (‘Chunkwang’ baechu; general spring cultivar, CB), two kinds of kimchi(ABK and CBK) prepared with AB and CB cultivar were fermented at 10°C for 10 days. Their fermentative characteristics and anti-proliferative activities against mouse carcinoma cell lines were investigated. General kimchi(CBK) showed mature pH on the 6th day of fermentation, whereas functional kimchi(ABK) reached pH on the 9th day. CBK also exhibited acidity of mature stage on the 6th day, but ABK reached mature acidity on the 9th day. Although ABK and CBK were salted in the same condition, ABK had lower salinity than CBK, throughout the fermentation time. The highest total bacterial and lactic bacterial counts of CBK showed on the 8th day of fermentation, but ABK showed the highest total bacterial and lactic bacterial counts on the 10th day. The texture of ABK was harder than CBK for fermentation time. This seems to be corrleated with the slower fermentation rate of ABK. ABK showed significantly higher anti-proliferative activity (54.6% cell viability of control) in B16BL6 at 1,000 μg/mL. ABK was also higher in anti-proliferative activity than CBK throughout the fermentation time. However, there was no significant difference in the anti-proliferative activity of ABK between the fermentation times. In conclusion, fermentation of ABK showed a better texture, due to the slow fermentation rate and more anti-proliferative activity against mouse carcinoma cell line than those of CBK.

Vegetable soup has been reported to have anti-inflammatory, anti-oxidative and anti-cancer effects. In this study, five kinds of vegetable soup were developed using a new manufacturing process and compositional changes in raw material, and anti-cancer and immuno-stimulatory activities were evaluated. Cytotoxicity tests based on MTT assay revealed that all vegetable soups had strong inhibitory effects against CT26 mouse colon cancer cells, with soups including Solomon’s seal being most effective based on comparison of IC50 values. Apoptosis in response to vegetable soup was occurred by 3-5 fold on cancer cells compared to normal cells. Mouse splenocytes increased by 266-541% in response to addition of vegetable soup in an in vitro proliferation experiment. In co-culture with splenocytes and CT26 cancer cells, splenocytes increased by more than 280% in every vegetable soup treatment, while cancer cells decreased by about 60% and cytokines such as IFN-γ and IL-12 were secreted from splenocytes in high levels only in response to vegetable soup including Solomon’s seal. In conclusion, all vegetable soups developed in this study had anti-cancer effects, and vegetable soup including Solomon’s seal showed the strongest anti-cancer and immuno-stimulatory effects. These results suggest that functionality of vegetable soup could be increased by changes in manufacturing processes and raw materials composition.

In this study, we investigated the effect of bisphosphonate on the osteoblastic differentiation of human dental stem cells (hDPSCs). In the first experiment, we evaluated the effect of bisphosphonate on the differentiation of hDPSCs into osteoblasts by alkaline phosphatase staining after culturing hDPSCs. As a result, on day 13, the osteogenic differentiation of hDPSC was suppressed at 5 μM in clodronate and 2 μM in zolendronate. In NBP, osteogenic differentiation is more suppressed. In second experiment, cytotoxicity and proliferation test, the cell proliferation (examined by MTT assay) was more suppressed as the concentrations of zolendronate were larger than those of alendronate and clodronate. Western blotting, a third experiment, was found that AKT phosphorylation was inhibited in cell signaling proteins involved in cell proliferation inhibition and death by bisphosphonate concentration. In human dental stem cells, bisphosphonates inhibit osteoblast differentiation, and this phenomenon is clearly observed in NBPs (zolendronate), and it has been found that it is related to AKT phosphorylation of cell signaling proteins.

척추동물의 혈구 세포의 특성은 활발히 연구되었지만, 곤충의 혈구 세포의 부피, 헤모글로빈의 양, 3차원 구조에대한 특성은 아직 잘 알려지지 않았다. 본 연구에서는, 주변에서 쉽게 구할 수 있는 곤충인 귀뚜라미를 대상으로하여 곤충의 혈구 샘플을 얻기 위한 실험 프로토콜을 제시하였으며, 광 회절 단층 촬영 (optical diffraction tomography)현미경 기술을 이용해 곤충의 혈구 세포의 3차원 구조를 정량적으로 측정하였다. 본 실험에서는 귀뚜라미의 혈구세포의 부피, 표면적, 구형도, 헤모글로빈 농도, 그리고 헤모글로빈 총량을 측정하였다. 측정된 다섯 개의 적혈구지표는 사람이나 쥐의 적혈구 지표와 유의미한 차이를 보였다.

최근 기능성과 친환경 화장품에 대한 관심이 증가하고 있으며, 이에 따라 안전하면서 효능이 우수한 식물 추출물을 활용한 소재 개발이 이루어지고 있는 실정이다. 따라서 본 연구에서도 주로 건강 기능성 소재로써 다양한 효능이 있는 것으로 알려진 그라비올라 추출물이 기능성 화장품 소재로써의 가 능성을 확인하고자 하였다. 그라비올라 추출물의 항산화 활성을 확인하고자 총 폴리페놀과 총 플라보노 이드 함량, DPPH radical 소거 활성을 측정하였고, HDF 세포에서의 세포 독성을 확인한 후 적정 농도 에서 HDF 세포에 과산화수소(H2O2)를 처리하여 산화적 스트레스에 대한 ROS 활성 억제 효과와 세포 보호 효과를 측정하였다. 본 실험 결과, 그라비올라 추출물은 항산화 지표가 되는 총 폴리페놀과 플라보 노이드의 100g당 26.6 mg(CA)/100g, 14.3 mg(CA)/100g의 높은 함량을 확인하였으며, 높은 radical 소 거 활성을 확인하였다. HDF 세포에 대한 세포 생존율을 측정한 결과, 모든 농도에서 유의한 세포 독성 이 나타나지 않았으며, 추후 100 μg/mL 농도에서 실험하였다. H2O2로 유도된 HDF 세포에 ROS 활성 억제를 측정한 결과, 농도 의존적인 ROS 활성 억제 효과를 확인하였고, H2O2를 4 시간, 24 시간, 48 시간 동안 처리 후 그라비올라 추출물의 세포 보호 효과를 측정한 결과, 25 μg/mL 농도에서 24시간까 지 89.92%의 높은 세포 보호 효과를 확인하였다. 이와 같은 결과를 통하여 그라비올라 추출물은 항산 화 활성이 우수하고, HDF 세포에 대한 독성이 거의 없으며, H2O2에 의해 발생하는 활성산소에 대한 효과적인 활성 억제 효과와 세포 보호 효과가 우수한 것으로 확인됨에 따라 항산화 및 세포 보호 효과 를 가진 다양한 기능성 소재로서의 가능성을 확인하였다.

본 연구에서는 고량(Sorghum nervosum) 70 % 에탄올 추출 후 항염증 및 UVB에 대한 세포 보호 효과를 검증하고자 하였다. 고량 추출물의 효능 평가는 세포생존율분석, 활성산소 측정, 항염증, COX-2 단백질 변화, UVB에 대한 세포 보호 효과를 수행하였다. 실험 결과, RAW264.7 대식세포, HaCaT 세포에서 고량추출물 모든 농도에서 97 %이상 세포 생존율을 확인하였다. 항염증 NO 생성 억 제능에서는 농도 의존적으로 저해 효과가 나타났으며, COX-2 단백질 발현량 역시 25, 50 ㎍/mL 농도 에서 유의하게(p＜.001) 저해되었다. UVB에 대한 세포 보호 효과로는 세포 내 활성산소종(ROS) 정량 분석 결과, 고량 추출물이 ROS 총 양 감소에 효과가 있음을 확인하였다. 연구결과를 종합적으로 고려해 볼 때, 고량추출물은 항염증 및 UVB에 대한 세포 보호 기능을 나타내는 화장품 원료로서의 개발 가능 성이 있다고 사료된다.

It is well-known that cultivated wild Panax ginseng has anti-inflammatory effect. However, a comparative study on cultivation period vs biofunctionality is currently lacking. In this study, 70% ethanol extracts of 3-years (yrs)-, 5-yrs-, or 7-yrs-old cultivated wild ginseng were evaluated for their inhibitory effects on RAW264.7 murine macrophages. Specifically, the production of pro-inflammatory cytokines (interleukin-6 [IL-6] and tumor necrosis factor-alpha [TNF-α]), the expression of surface proteins (CD80, CD86, and MHC-II), and the phagocytic properties were investigated. RAW264.7 cells were induced by 500 ng/mL of lipopolysaccharide (LPS) and treated with 0.1, 1, and 10 ppm of samples. LPS-induced IL-6, TNF-α and surface proteins in all samples were downregulated in a dose-dependent manner. Both IL-6 and TNF-α were significantly reduced at 10 ppm of the 7-yrsold sample compared to 10 ppm of 3-yrs- and 5-yrs-old samples. CD80 and CD86 were also reduced at 10 ppm of all samples, and there was no difference among samples. The phagocytosis has no difference except in 10 ppm of 3 yr-old sample. The results suggest that cultivated wild ginseng extract has anti-inflammatory effect without decreasing phagocytosis.

In most mammals, metaphase II (MII) oocytes having high maturation promoting factor (MPF) activity have been considered as good oocytes and then used for assisted reproductive technologies including somatic cell nuclear transfer (SCNT). Caffeine increases MPF activity in mammalian oocytes by inhibiting p34cdc2 phosphorylation. The objective of this study was to investigate the effects of caffeine treatment during in Vitro maturation (IVM) on oocyte maturation and embryonic development after SCNT in pigs. To this end, morphologically good (MGCOCs) and poor oocytes (MPCOCs) based on the thickness of cumulus cell layer were untreated or treated with 2.5 mM caffeine during 22-42, 34-42, or 38-42 h of IVM according to the experimental design. Caffeine treatment for 20 h during 22-42 h of IVM significantly inhibited nuclear maturation compared to no treatment. Blastocyst formation of SCNT embryos was not influenced by the caffeine treatment during 38-42 h of IVM in MGCOCs (41.1-42.1%) but was significantly improved in MPCOCs compared to no treatment (43.4 vs. 30.1%, P<0.05). No significant effects of caffeine treatment was observed in embryo cleavage (78.7-88.0%) and mean cell number in blastocyst (38.7-43.5 cells). The MPF activity of MII oocytes in terms of p34cdc2 kinase activity was not influenced by the caffeine treatment in MGCOCs (160.4 vs. 194.3 pg/ml) but significantly increased in MPCOCs (133.9 vs. 204.8 pg/ml). Our results demonstrate that caffeine treatment during 38-42 h of IVM improves developmental competence of SCNT embryos derived from MPCOCs by influencing cytoplasmic maturation including increased MPF activity in IVM oocytes in pigs.