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.

Tissue engineering (TE) has been developed to create functional organs and tissue by combining 3D matrix and cells in vitro. Vascularization and angiogenesis are utmost important for supply of nutrients and oxygen in tissue engineered organs. The present study was performed to isolate and characterize primary endothelial cells (EC) from aorta of alpha 1, 3-enzyme galactosyltransferase knock out (GalT KO) pig, to minimize immune rejection and analyze body immune system for future xenotransplantation studies. Isolation of primary EC from aorta were performed by incubation with dispase for 8-10 min at 37°C. Primary EC were cultured in EC growth medium on different extra cellular matrix (ECM), either collagen or gelation. Primary EC exhibits morphological characteristics and showed positive expressions of EC specific marker proteins i.e. PECAM1, KDR and VWF despite of their ECM surface; however, on collagen based surface they showed increase in mRNA level analyzed by qPCR. Primary EC cultured on collagen were sorted by flow cytometer using KDR marker and cultured as KDR positive cells and KDR negative cells, respectively. KDR positive cells showed dramatically increased in PECAM1 and VWF level as compared to KDR negative cells. Based on the above results, primary EC derived from GalT KO are successfully isolated and survived continuously in culture without becoming overgrown by fibroblast. Therefore, they can be utilize for xeno organ transfer, tissue engineering, and immune rejection study in future.

After being subjected to different cooking methods, small black beans (Rhynchosia nulubilis) were investigated in order to assess the effects of the retained bioactive compounds. Using uncooked, pan broiled, boiled, steamed, and pressure cooked beans, the inhibitory effects of MCF-7 cell migration were evaluated at protein concentrations of 40, 160, and 640 μm/mL, using the Boyden's chamber assay. All protein concentrations (40, 160, and 640 μm/mL) of pan broiled beans showed significant reduction (59.83, 32.48, and 21.37%, respectively) in the rate of cell migration to the lower chambers (p-value less than 0.001). Estimated cell migration rates correlated to the exponential decay between experimentally measured cell migration rates and converted samples. The range of estimated cell migration rate for each 100 mg/mL of cooked sample was as follows: pan broiled (21.16%), boiled (22.48%), steamed (22.48%), pressure cooked (29.52%), and uncooked (35.03%) beans. Our study indicated that selective modifications of cooking methods for small black beans, such as pan broiling, ameliorated the inhibitory effects of MCF-7 cell migration. This suggests that optimized cooking methods increase the nutritional contents of the cooked food.

Fludarabine, a chain terminating anti-cancer drug, is a purine analogue that causes DNA strand breaks in normal cells. In this study, we determined if A. melanocarpa and Korean red ginseng extract mixture reduce cytotoxicity of fludarabine. Treatment of HaCaT cells with 10 μM of fludarabine for 24 hours decreased cell viability and increased DNA strand breaks. Treatment of A. melanocarpa and Korean red ginseng extract mixture for 24 hours increased cell viability as compared with single extract treatment. The protective effect of these extracts on cell activity increased in a concentration-dependent manner. DNA strand breaks induced by fludarabine decreased as concentration of extract mixture increased. p-H2AX level, a marker of DNA strand breakage, decreased depending on the concentration of extract mixture. The effect of mixed extract of A. melanocarpa and Korean red ginseng on DNA damage is due to the anti-oxidative effect of A. melanocarpa and signal transmission through glucocorticoid receptor upon binding of saponin of Korean red ginseng.

Connective tissue growth factor (CTGF, CCN2) is one of the multi-functional secreted proteins which belong to CCN family of cysteine-rich growth factors. CTGF is known to have pivotal roles in embryonic endochondral ossification but its role in relevance to periodontitis is never been determined. To identify new molecular mediators associated with periodontitis-induced bone resorption, we have analyzed publicly available GEO database and found the markedly augmented CTGF mRNA expression in periodontitis gingival tissues. The existence of CTGF significantly enhanced mature osteoclasts survival which accompanied by reduction in TUNEL-positive nuclei and PARP cleavage. These results may provide another line of evidence the CTGF mediated prolonged osteoclast survival and subsequent increased bone resorption in the periodontitis patients.

King oyster mushroom (Pleurotus eryngii), an improved species of oyster mushroom, is a popular ingredient in Asian cuisine. Spleen cells were treated with various concentrations (0, 5, 10, 50, 100, 250, 500, and 1,000 μg/mL) of king oyster water extracts (KOWE); then, the proliferation of the cells was measured 24, 48, and 72 h after each treatment. Also, type 1 T helper cytokine productions (TNF-α, IFN-γ, and IL-2) were measured in activated macrophage by KOWE in seven concentrations. Under the condition of its 50, 100, 250, and 1,000 μg/mL for 48 h, the proliferation of cells was increased. However, there was no significant fluctuation in the spleen cells proliferation for 24 and 72 h-long KOWE exposure. To determine cytokine (TNF-α, IFN-γ, IL-2) productions of type 1 T helper cells, macrophage was stimulated by KOWE for 48 h. Treatment of KOWE gave a rise to the levels of TNF-α and IFN-γ, but not in that of IL-2 productions. These results suggest that king oyster mushroom water extracts may be beneficial for enhancing immune functions in its high concentration.

Plantago asiatica L., observed frequently in East Asia, is a known herb used in traditional medical remedies several studies report that P. asiatica L has anti-inflammatory and antioxidant effects. To determine the production of cytokines (IL-2, IFN-γ, and TNF-α) induced by lipopolysaccharide (LPS) and non-LPS-stimulated macrophages, an ELISA assay was conducted using cytokine kits. Mice splenocytes were cultured for 48 h with various concentrations of P. asiatica L. (5, 10, 50, 100, 250, 500, and 1,000 μg/mL) or with mitogens (ConA or LPS). P. asiatica L. increased the proliferation of mice splenocytes, especially under the condition of its concentration ranging from 250 to 1,000 μg/mL. In addition, Plantago asiatica L. notably induced cytokine production of (IL-2, IFN-γ, and TNF-α) at its concentration of 250~500 μg/mL. These results suggest that supplementation with P. asiatica L. water extracts may play a potential role in enhancing immune function by mediating splenocyte proliferation and cytokine production through its anti-inflammatory activit.