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본 연구는 와송 유기용매 추출물중 DCM층을 이용하여 지방세포로 유도된 3T3-L1세포에 대한 지질대사 개선효과를 확인하는 연구이다. 세포독성을 확인하기 위하여 MTS-assay를 이용하여 6가지 분획의 와송 유기용매 추출물(EtOH, Hexane, DCM, EToAc, BuOH, H2O)에 대한 독성검사를 실시하여 세포에 대한 안정성을 확인하였다. 그 결과 DCM추출물이 전 농도에 걸쳐 안정성이 있음을 확인하고, 지질대사 개선효과를 확인하기 위하여 DCM층을 사용하였다. 우선 지질대사 개선효과를 확인하기 위하여 지질 배출능을 측정하였다. 지방세포로 유도된 3T3-L1세포에 대하여 DCM추출물을 처리한 후 oil-red O 염색을 실시하여 지질 축적량을 평가한 결과 지질 배출능이 개선된 것을 확인할 수 있었다. 또한 지질 배출능 분석을 위하여 지질수송 단백질인 ABCA1, ABCG1의 mRNA발현을 real-time PCR로 확인한 결과 DCM추출물 처리 시 유의적인 발현 증가가 나타났음을 확인 하였다. 따라서 본 연구를 통하여 와송 DCM추출물이 3T3-L1 지방세포에 대하여 지질대사 개선효과가 있음을 확인하였다.

The immunomodulatory effects of mesenchymal stem cells (MSCs) are an important mediator of their therapeutic effects in stem cell therapy and regenerative medicine. The regulation mechanism of MSCs is orchestrated by several factors in both intrinsic and extrinsic events. Recent studies have shown that the dynamic expression of cytokines secreted from MSCs control T cell function and maturation by regulating the expression of FoxP3, which figures prominently in T cell differentiation. However, there is no evidence that placenta-derived mesenchymal stem cells (PD-MSCs) have strong immunomodulatory effects on T cell function and maturation via FoxP3 expression. Therefore, we compared the expression of FoxP3 in activated T cells isolated from peripheral blood and co-cultured with PD-MSCs or bone marrow-derived mesenchymal stem cells (BM-MSCs) and analyzed their effect on T cell proliferation and cytokine profiles. Additionally, we verified the immunomodulatory function of PD-MSCs by siRNA-mediated silencing of FoxP3. MSCs, including PD-MSCs and BM-MSCs, promoted differentiation of naive peripheral blood T cells into CD4+CD25+FoxP3+ regulatory T (Treg) cells. Intriguingly, the population of CD4+CD25+FoxP3+ Treg cells co-cultured with PD-MSCs was significantly expanded in comparison to those co-cultured with BM-MSCs or WI38 cells (p

Orostachys japonicus (O. japonicus) is utilized as a traditional medicine for patients with various diseases. This study investigated the effect of the ethyl acetate fraction from O. japonicus extract (OJE) on the growth inhibition of OVCAR‐3 human ovarian cancer cells demonstrated to inhibit cell growth and arrest the cell cycle in OVCAR‐3 cells by blocking the sub‐G1 phase and decreasing cyclin E1/CDK2 expression. Cell cycle arrest was connected to the increased expression of the cell cycle regulating factors p53 and p21. Apoptosis was initiated through the intrinsic pathway by up‐regulating the expression of the Bcl‐2/Bax ratio and down‐regulating the expression of pro‐caspase‐3. Furthermore, OJE treatment elicited p‐p38 activation and p‐ERK1/2 inhibition. In conclusion, our results demonstrated that OJE reduced the growth of OVCAR‐3 human ovarian cancer cells mediated by arrest of the cell cycle and regulation of MAPK signaling pathways.

Orostachys japonicus (O. japonicus) was extracted with ethanol (EtOH) and sequentially separated with organic solvents, including n‐hexane (Hex), dichloromethane (DCM), ethyl acetate (EtOAc), n‐butanol (BuOH), and water (H2O). All the fractions were confirmed for anti‐inflammatory activity in an inflammatory condition. The DCM fraction showed the highest anti‐inflammatory ability. Here, we examined the effect of DCM fraction and investigated the intracellular signaling pathways in LPS‐stimulated RAW 264.7 macrophage cells. The DCM fraction significantly inhibited the mRNA levels of pro‐inflammatory mediators and cytokines including iNOS, COX‐2, IL‐1β, IL‐2, IL‐6, and IP‐10 in LPS‐stimulated cells. Also, the treatment of DCM fraction excellently reduced the expression of the proteins of AP‐1 (phospho‐c‐Jun and phospho‐c‐Fos) and phospho‐IRF3 as transcription factors. As a result, it suppressed LPS‐induced inflammatory mediator and cytokines via inhibition of transcription factors. In conclusion, our data demonstrated that DCM fraction has a strong anti‐inflammatory activity that improves the inflammatory state., We examined the effect of DCM fraction and investigated the intracellular signaling pathways in LPS‐stimulated RAW 264.7 macrophage cells. Our data demonstrated that DCM fraction has a strong anti‐inflammatory activity that improves the inflammatory state.

Salicornia herbacea L. (S. herbacea) is an annual herbaceous plant of Chenopodiaceae. It grows in groups on the coast or mud flat of Korea is known to be rich in minerals. S. herbacea has potent anti-cancer, antioxidant, anti-obesity, bowel function improvement. However, pharmacological mechanisms of S. herbacea extract (SHE) remain poorly understood. The aim of this study was to investigate the potential acute toxicity of SHE in ICR mice administered a single oral dose of 0, 500, 1,000, and 2,000 mg/kg by gavage. After administration of the extract, signs of toxicity were observed every day for 14 days. No mortality, abnormal clinical signs, body weight, organ weight or pathological changes were observed compared to a control group, and there were no differences in the body weights of the control and treatment groups. Biological serum activities and histological tests were not significantly changed in the treatment group compared to the control group. Especially, treatment of SHE was significantly decreased of total cholesterol and triglyceride levels. These results indicated that a single oral administration of SHE does not exerts any toxic effects at a dose of 2,000 mg/kg and that the LD50 of SHE is greater than 2,000 mg/kg. Accordingly, SHE appears to have potential in various functional agents of foods, without toxicity.

In order to study the effect of the receptor protein (SeaR), which is isolated from Saccharopolyspora erythraea, we introduced the SeaR gene to Streptomyces virginiae as host strains. An effective transformation procedure for S. virginiae was established based on transconjugation by Escherichia coli ET12567/pUZ8002 with a φC31-derived integration vector, pSET152, which contained int, oriT, attP, and ermEp * (erythromycin promotor). Therefore, the pEV615 was introduced into S. virginiae by conjugation and integrated at the attB locus in the chromosome of the recipients by the φC31 integrase (int) function. Transformants of S. virginiae containing the SeaR gene were confirmed by PCR and transcriptional expression of the SeaR gene in the transformants was analyzed by RT-PCR, respectively. And, we examined the production time of virginiamycins in the culture media of both the transformants and the wild type. The production time of virginiamycins in the wild type and transformants was the same. When 100 ng/㎖ of synthetic VB-C6 was added to the state of 6 or 8 hour cultivation of wild type and transformants, respectively, the virginiamycins production was induced, meaning that the virginiamycins production in the wild type was detected 2 h early than transformants. From these results, SeaR expression was also affected to virginiamycins production in transformants derived from S. virginiae. In this study, we showed that the SeaR protein worked as a repressor in transformants.

Lactobacillus spp.같은 유산균은 탄수화물을 발효하여 ATP를 생성하며, 시판되는 요구르트 생산에 이용되고 있다. Lactobacillus spp.는 장관계 건강에 유익한 균으로, 특히 Lactobacillus acidophilus를 함유하는 요구르트는 대장암 초기단계를 예방하는 효과가 있는 것으로 나타나 관심이 모아지고 있다. 본 연구에서는 ...

Secondary metabolism in actinomycetes has been known to be controlled by a small molecule, -butyrolactone autoregulator, the binding of which to each corresponding receptor leads to the regulation of the transcriptional expression of the secondary metabolites. We expected that expression of an autoregulator receptor or a pleiotropic regulator in a non-host was to be gained insight of effective production of new metabolic materials. In order to study the function of the receptor protein (seaR), which is isolated from Saccharopolyspora erythraea, we introduced the seaR gene to Streptomyces coelicolor A3(2) as host strains. An effective transformation procedure for S. coelicolor A3(2) was established based on transconjugation by Escherichia coli ET12567/pUZ8002 with a -derived integration vector, pSET152, which contained int, oriT, attP and (erythromycin promotor). Therefore, the pEV615 was introduced into S. coelicolor A3(2) by conjugation and integrated at the attB locus in the chromosome of the recipients by the integrase (int) function. Exconjugant of S. coelicolor A3(2) containing the seaR gene was confirmed by PCR and transcriptional expression of the seaR gene in the transformant was analyzed by RT-PCR. In case of S. coelicolor A3(2), a phenotype microarray was used to analyze the phenotype of transformant compared with wild type by seaR expression. After that, in order to confirm the accuracy of the results obtained from the phenotype microarray, an antimicrobial susceptibility test was carried out. This test indicated that sensitivity of the transformant was higher than wild type in tetracycline case. These results indicated that some biosynthesis genes or resistance genes for tetracycline biosynthesis in transformant might be repressed by seaR expression. Therefore, subsequent experiments, analysis of transcriptional pattern of genes for tetracycline production or resistance, are needed to confirm whether biosynthesis genes or resistance genes for tetracycline are repressed or not.

Background: The etiology of parkinsonism involves gene and environment interaction, but the identity of most causal factors remains unknown. Investigating the distribution and characteristics of the condition is important for identifying further etiologic factors and planning public health policies. Objectives: To validate various known risk factors of Parkinsonism and to establish basic information to formulate public health policy by using an 11 year followup cohort model. Methods: This population based nationwide study was performed using the National Health Insurance Database of reimbursement claims of the Health Insurance Review and Assessment Service of South Korea data on regular health check-ups in 2003 and 2004, with 11 years` followup. Results: We identified 7,746 patients with Parkinsonism. Old age, hypertension, diabetes, depression, anxiety, taking statin medication, high body mass index, nonsmoking, nonalcohol drinking, and low socioeconomic status were each associated with an increase in the risk of Parkinsonism (fully adjusted Cox proportional hazards model: hazard ratio 1.259, 95% confidence interval 1.194~61485;1.328 for hypertension, hazard ratio 1.255, 95% confidence interval 1.186~61485;1.329 for diabetes, hazard ratio 1.554, 95% confidence interval 1.664~61485;1.965 for depression, hazard ratio 1.808, 95% confidence interval 1.462~61485;1.652 for anxiety, and hazard ratio 1.157, 95% confidence interval 1.072~61485;1.250 for taking statin medication). Conclusions: In our study, old age, depression, anxiety, and nonsmoker status were found to be risk factors of Parkinsonism, concordant with previous studies. However, sex, hypertension, diabetes, taking statin medication, nondrinking of Alcohol, and lower socioeconomic status have not been described as risk factors previously and need further verification in future studies.

Department of Biomedical Laboratory Science, Inje UniversityAbstract This study investigated the effect of insoluble dietary fibers extracted from Salicornia herbacea L. (S.herbacea) on the improvement of intestinal function in rats. Sprague-Dawley rats were fed on diet containing 5% and 10%S. herbacea dietary fiber (SHDF) for four weeks. Rats receiving the SHDF diet showed a significant decrease in theirtriglyceride levels and an increase in HDL-cholesterol levels. In addition, compared with the control group, the SHDFgroup showed a significant increase in the total quantity of the feces and its moisture content. The intestinal transit timeof the feces was also shorter in this group. The pH of the feces decreased in all the other experimental groups. Particularly,the bile acid content of the feces and the thicknesses of the mucus layers showed significant recovery on SHDF intake.These results suggest that dietary fiber isolated from S. herbacea has a marked effect on the improvement of bowelfunction in rats with loperamide (2 mg/kg)-induced constipation.Keywords: insoluble dietary fiber, Salicornia herbacea L., constipation, bowel function

Department of Biomedical Laboratory Science, Jungwon University , Goesan, Chungbuk 367-700, KoreaReceived August 18, 2014/Revised September 12, 2014/Accepted October 14, 2014A yellow-colored pigment is found in turmeric, or Curcuma longa L. (Zingiberaceae), a perennial herb distributed mainly throughout tropical and subtropical regions. C. longa has potent antiviral, anti-mutagenic, anti-inflammatory, anticancer, and antioxidant properties. However, pharmacological mechanisms of ethanol extract derived from C. longa remain poorly understood. The aim of this study was to investigate the potential acute toxicity of C. longa (Curcuma longa L.) extract in BALB/c mice administered a single oral dose of 0, 20, 200, and 2,000 mg/kg by gavage. After the administration of the agent, signs of toxicity were observed every hour for the first 6 hr and every day for 14 days. No mortality, abnormal clinical signs, or pathological changes were observed compared to a control group, and there were no differences in the body weights of the control and treatment groups. Biological serum activities were not significantly changed in the treatment group compared to the con-trol group. These results indicate that a single oral administration of C. longa extract does not exert any toxic effects at a dose of 2,000 mg/kg body weight and that the LD

Orostachys japonicus (Crassulaceae) is referred to as Wa-song in Korea. It is used as an anti-inflammatory, antifebrile, hemostatic, and anti cancer agent, and as an antidote.The purpose of this study was to evaluate the acute toxicity of the ethyl acetate fraction of O. japonicus (OJE) after the oral administration in Balb/c mice of both sexes.Mice were oral administered a single doses of 500, 1000, and 2000 mg/kg of body weight and were monitored for 14 d. Biochemical parameters [aspartate amino transferase (AST), alanine amino transferase (ALT), alkaline phosphatase (ALP), total protein (TP), globulin (GB), total cholesterol (TC), triglyceride (TG), blood urea nitrogen (BUN), and creatinine (CR)] and histopathological examination of liver were performed.No animals died and no toxic changes were observed in clinical signs, body weight, and organ weight. The LD50 of orally administered OJE was higher than 2000 mg/kg/d in both sexes. No toxicological findings were found in biochemical parameters. In histophathological examination, neutrophilic infiltration was observed at a dose of 2000 mg/kg group in both sexes. These finding suggest that oral administration of OJE does not produce acute toxicity. Therefore, these results could provide satisfactory preclinical evidence of safety to launch clinical trials on standardized formulation of OJE to be a biohealth product.

Orostachys japonicus shows various biological activities. However, the molecular mechanisms remain unknown in LPS-stimulated macrophages. Here, we investigated the anti-oxidizing effect of the dichloromethane (DCM) and hexane fractions from O. japonicus (OJD and OJH) against oxidative stress in RAW 264.7 cells stimulated by LPS. OJD and OJH significantly increased the expression of heme oxygenase-1 (HO-1) in a dose- and time-dependent manner. Additionally, it was found that the expression of HO-1 was stimulated by Nrf2 activated via degradation of Keap1. ERK and p38 inhibitors repressed HO-1 induced by OJD and OJH in LPS-stimulated cells, respectively. In conclusion, these results suggest that OJD and OJH may block oxidative damage stimulated by LPS, via increasing the expression of HO-1 and Nrf2, and MAPK signaling pathway. [BMB Reports 2014; 47(2): 98-103]

The molecular mechanisms of the ethylacetate (EtOAc) fraction from Orostachys japonicus (OJE) (including gallic acid, kaempferol, and quercetin) for anti-cancer activities in HepG2 cells were investigated. Apoptosis was detected using morphological observation of nuclear changes and investigation of phosphatidylserine exposure at the cytoplasmic membrane using FITC-Annexin V/PI staining. Activities of the apoptotic factors bcl-2, bax, cytochrome c, pro-caspase-3, 8, and 9, as well as MAPKs levels, were measured using western blotting. Some morphologic features of apoptosis were identified using confocal microscopy. OJE caused the expression of apoptotic proteins to change, as evidenced by an increased bax/bcl-2 ratio and increased expression of cytochrome c, and decreased expressions of pro-caspase-3, 8, and 9. After HepG2 cells were exposed to OJE, expressions of p-JNK and p-ERK1/2 increased in a dose dependent manner. OJE exhibits anti-cancer activity via apoptosis induction through a mitochondria dependent signaling pathway in HepG2 cells.

Cancer rates are increasing dramatically, and there is currently a strong emphasis on identifying biologically active substances with anti-cancer activity from traditional herbs, as these are thought to have less adverse side-effects than conventional chemotherapy. Here, we examined the effects of extracts of Orostachys japonicus A. BERGER (O. japonicus) on cancer cell proliferation and apoptosis, and investigated the underlying signaling pathways. Dried powdered O. japonicus was extracted with 95% ethyl alcohol and fractionated with a series of organic solvents, including n-hexane (hexane), dichloromethane (DCM), ethylacetate (EtOAc), n-butanol (BuOH), and water (H(2)O). These extracts were tested for anti-cancer activity on a range of cancer cells; of all these, the EtOAc soluble fraction showed the highest anti-cancer activity, which was most marked in AGS human gastric cancer cells. The EtOAc fraction inhibited the proliferation of AGS cells in a dose-dependent and time-dependent manner, by inducing apoptosis and cell cycle arrest, as evidenced by 4,6-diamidino-2-phenylindole (DAPI) staining, annexin V-fluorescein isothiocyanate staining, propidium iodide-labeling, and DNA fragmentation assays. Western blot analysis revealed that p53 and cleaved caspase-3 proteins were up-regulated, and B cell lymphoma-2 (bcl-2) protein and pro-caspase-3 were down-regulated, but bcl-2 associated x protein (bax) protein was not regulated, in response to treatment of AGS cells with the EtOAc fraction. However, the changes of pro-caspase-3 and cleaved caspase-3 could be abolished by the pan-caspase inhibitor Z-VAD-FMK. These results suggest the EtOAc fraction from O. japonicus has substantial anti-cancer activity in human gastric cancer cells.

Ethnopharmacological relevance Orostachys japonicus A. Berger (O. japonicus) is known to reduce the risk of many diseases. Aim of the study We investigated the anti-inflammatory effects of the dichloromethane (DCM) fraction from O. japonicus (OJD) in LPS-stimulated RAW 264.7 cells. Materials and methods NO was measured using the Griess method. Key pro-inflammatory cytokines and mediators including IL-1β, TLR4, iNOS, and COX-2; 2 important pro-inflammatory transcription factors, NF-κB p65 and IκBα; and MAPKs such as ERK1/2, JNK, and p38 were analyzed by Western blotting. Results OJD significantly inhibited NO production, IL-1β, TLR4, iNOS, and COX-2 expression in LPS-stimulated cells. Additionally, it inhibited LPS-induced NF-κB p65 activation via inhibition of IκBα phosphorylation. Furthermore, phosphorylation of p38 and JNK was suppressed by OJD in a dose-dependent manner in the LPS-stimulated RAW 264.7 cells. Conclusions Our data suggest that OJD inhibits the inflammatory response via suppression of NF-κB activation and MAPK signaling.

This study was designed to investigate the potentially protective effects of Curcuma longa Linn. extract (CLE) on carbon tetrachloride (CCl4)-induced hepatotoxicity in rats. Male Sprague-Dawley rats were pretreated with 50 or 100mg/kg of CLE or 100mg/kg of butylated hydroxytoluene (BHT) for 14 days before CCl4 administration. In addition, the CLE control group was pretreated with 100mg/kg CLE for only 14 days. Three hours after the final treatment, a single dose of CCl4 (20mg/kg) was administrated intraperitoneally to each group. After the completion of this phase of the experiment, food and water were removed 12 h prior to the next step. The rats were then anesthetized by urethane and their blood and liver were collected. It was observed that the aspartate aminotransferase and alanine aminotransferase activities of the serum, and the hepatic malondialdehyde levels had significantly decreased in the CLE group when compared with the CCl4-treated group. The antioxidant activities, such as superoxide dismutase, catalase, and glutathione peroxidase activities, in addition to glutathione content, had increased considerably in the CLE group compared with the CCl4-treated group. Phase II detoxifying enzymes, such as glutathione S-transferase, were found to have significantly increased in the CLE group as opposed to the CCl4-treated group. The content of Nrf2 was determined by Western blot analysis. Pretreated CLE increased the level of nuclear translocated Nrf2, and the Nrf2 then increased the activity of the antioxidant and phase II detoxifying enzymes. These results indicate that CLE has protective effects against CCl4-induced hepatotoxicity in rats, via activities of antioxidant and phase II detoxifying enzymes, and through the activation of nuclear translocated Nrf2.