Background : Mesenchymal stem cells (MSCs) are present in most of the tissue matrix, taking part in their regeneration when injury or damage occurs. The aim of this study was to investigate the presence of cells with pluripotential characteristics in human subchondral bone and the capacity of these cells to differentiate to osteoblast. Methods : Human subchondral bone were digested with collagenase. Isolated cells were cultured with a-MEM, 15% FBS, 10-8M dexamethasone and 50 ng/mL ascoric acid. Cells from 0 day(isolated cells), 7 day (first subculture) and 14 days (third subculture) were used to carry out phenotypic characterization experiments flowcytometry analysis with 11 monoclonal antibodies) and osteogenic differentiation experiments. Osteogenic differentiation of cells was assessment by quantification of bone extracellular matrix components by following analysis: alkaline phosphatase(ALP) stains to detect ALP activity, RT-PCR and western blot to detect osteocalcin (OCN), osteopontin (OPN) and type I collagen(Col I), and Alizarin red stains to detect calcium deposition. Results : Flowcytometry analyses showed that in our population more than 98% of cells were positive for MSC markers: SH-2(CD105, 99%), CD29 (95%), CD73 (95%). Cells were negative for hematopoietic markers (CD11b, CD34, and CD45). Furthermore, cells showed positive stain to multipotent markers such as CDl17 (c-kit) (15.1%), and CD166 (74.9%), and cell adhesion molecules such as CD54 (78.1%) and CD106 (63.5%). The osteogenic specific marker analyses showed that the culture of these cells for 7 and 14 days stimulates ALP, OCN, OPN and Col I synthesis by RT-PCR and Western blot analysis. Also, after 14 days in the culture of MSCs induces mineralization by Arizarin red stain. Conclusion : In this work, we demonstrated a new and efficient method for osteoblastic differentiation of human subchondral bone stem cells. As MSCs takes part in reparative processes of adult tissues, these cells could play an important role in osteogenesis.
Curcumin has diverse anticancer activities that lead to tumor growth inhibition of cancer cells and induction of apoptosis. Curcumin is involved in the regulation of multiple genes via transcription factors including NF-${\kappa}B$, STATs, AP1, and SP. Notch signaling plays critical roles in maintaining the balance between cell proliferation, differentiation and apoptosis, and thereby may contribute to the development of various cancers involving breast cancer. This study was to investigate the effects of curcumin on Notch1 gene expression and to explore the underlying mechanism. Here, we found that curcumin decreased the levels of Notch1 mRNA and protein in MDA-MB-231 human breast cancer cells, along with the downregulation of Sp family genes (Sp1, Sp2, Sp3, and Sp4). The repressive effect of curcumin on Notch1 gene transcription was confirmed by performing Notch1 promoter-driven reporter assay and three Sp-binding sites were identified on Notch1 promoter that may act as curcumin-respose elements. Moreover, treatment with mitramycin A, a specific Sp inhibitor, decreased the levels of Notch1 mRNA and protein in human breast cancer cells. Taken together, our results indicate that Notch1 gene expression is downregulated by curcumin, at least in part, through the suppression of Sp family, which may lead to apoptosis in human breast cancer cells.
The present study was undertaken to investigate the effects of different nitrogen source and rate on the growth of Fusarium oxysporum f. vasinfectum which is known to be a noticeable fungus causing the wilt disease of both sesame and cotton in Korea. From the results of this study, It was known that different N source and rate markedly affect the growth of Fusarium oxyspsrum f. vasinfectum Among four N sourses were used in this study, nitrate-N and urea-N were appropriate N source for the growth of fungus. Above all, nitrate N was the best N source because it is utilized in more extensive range of concentration in comparison with the other N source by the fungus, On the other hand, ammonia-N is of little avail for the growth of the fungus because of the formation of unusual colonies with wavy margin and bead-like mycelial cells in addition to marked reduction of mycelial growth and B sporulation of the fungus irrespective of concentration. Judging from the formation of such an abnormal colony and bead-like mycelial cell which is known to be a characteristic of 'staling-type' growth of fungi, the effect of ammonia-N on the growth of Fusarium oxysporum f. vasinfectum is similar to that of phenoxy componnds on some other fungi previously investigated by some workers. Ammonium and nitrate also was not considered to be an appropriate source for the growth of the fungus because of the formation of colonies with slight wavy margin and appreciable reduction of mycelial growth and sporulation in higher concentration than 50meq. , although much or less masking of the irregularity of colony occurs. Therefore, ammonia N alone or any other N combined with ammonia N is of little avail for the growth of Fusarium oxysporum f. vasinfectum.
Kim, Mi-Jin;Im, Kyung-Ran;Jeong, Taek-Gyu;Yoon, Kyung-Sup;Choi, Sang-Won
Journal of the Society of Cosmetic Scientists of Korea
/
v.35
no.2
/
pp.143-149
/
2009
Several hydroxycinnamic acid derivatives, p-coumaric acid, ferulic acid, N,N'-dicoumaroylputrescine (DCP), N-p-coumaroyl-N'-feruloyl-putrescine (CFP), and N,N'-diferuloylputrescine (DFP) were isolated and purified from corn bran. To develop the skin whitening agent, we investigated the effects of hydroxycinnamic acid derivatives from corn bran, on melanogenesis. CFP and DFP inhibited melanin synthesis in a dose dependent manner up to 44.7 ${\pm}$ 6.0 %, and 58.5 ${\pm}$ 3.1 % at a concentration of 50 ${\mu}g/mL$, respectively. The intracellular tyrosinase activity decreased about 42.5 ${\pm}$ 14.6 %, and 9.0 ${\pm}$ 4.4 % at a concentration of 50 ${\mu}g/mL$ of CFP and DFP, respectively. Our results suggest that inhibitory effects of hydroxycinnamic acid derivatives on melanogenesis are due to the inhibition of the intracellular tyrosinase activity. These results indicate that these hydroxycinnamic acid derivatives from corn bran may be potential natural skin whitening agents.
The cycle of the seminiferous epithelium and development of spermatids of Apodemus speciosus peninsulae were observed using a light microscope. On the basis of developing spermatocyte and spermatid, the cycle of the seminiferous epithelium was divided into 9 stages. Type Ad spermatogonia were appeared in all stages ($I{\sim}IX$). The Ap, In, and B types of spermatogonia were appeared from stage I, II and III, and IV, respectively. In prophase of first meiosis, the leptotene spermatocytes appeared from stage V and VI, zygotene spermatocytes from stages I, II, VII, VIII, and IX, pachytene spermatocytes from stage III to VII, diplotene spermatocytes in the stage VIII, and secondary spermatocytes in stage IX. On the basis of morphology of spermatid head, developing of nuclear and acrosome and the morphological change of cytoplasm, the developing of spermatids was divided into 12 steps. Considering all the results, A. s. peninsulae displayed very similar result with A. agrarius coreae that is allied species when compare correct characters developing of spermatids with spermatogonia and appearance time of the spermatocyte. Appearance time of the same cell and number of spermatogonial generation was thought that characters of the species, and information may be useful in identifying the species.
Lee, Kwang Ho;Cha, Mi Young;Chung, Woo Yang;Bae, Hyeun-Jong;Kim, Yoon Soo
Journal of the Korean Wood Science and Technology
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v.37
no.6
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pp.566-577
/
2009
The modification of wood color occurs rapidly during the service period at indoor. It is crucial to investigate the characteristics of color change, chemical and microscopical modification of wood at indoor. Wood products made of Japanese cedar at different years were used for this work. The tests were performed in order to evaluate the degree of color change of wood surface, breakpoint of brightness from surface to inside of wood, chemical analysis with FT-IR, and microscopical characteristics using the LM and TEM. Surface color of cedar wood stored indoor were rapidly changed at early stage, particularly ${\Delta}a$ (yellow), and ${\Delta}b$ (red) values were steeply decreased for one year old indoor wood, ${\Delta}L$ (white) value was dropped until 5 years old indoor wood compared with control sample. Decrease of peaks related to polysaccharide and lignin was noticed, especially, lignin was severely degraded. Although degradation of cell wall limited only to surface layers of indoor wood, degradation pattern of indoor wood showed similar degradation pattern to natural weathering of wood during outdoor weathering or wood behavior under artificial UV irradiation.
Objectives: We aimed to identify the dose-dependent inhibitory effects of Yukmijihwang-tang-Hap-Sabaek-san(YMHSB) and Root cortex of Morus alba L.(RCM) on the mRNA expression of Interieukin(IL)-6, IL-S, granulocyte macrophage colony stimulating factor(GM-CSF) involved in the asthma model. Methods: In this study BEAS-2B cell lines, human epithelial cells, were used. These cells were stimulated by tumor necrosis $factor(TNF)-{\alpha},\;IL-1{\beta}$ and histamine for artificial inflammatory expression. ${\beta}-actin$ messenger RNA(mRNA) was used for the internal standard. After each 24 hours of the YMHSB and RCM treatment, total cellular RNAs were collected by treating RNA zol directly on the living cells. Then the transcriptional activities of IL-6, 8 and GM-CSF were measured by RT-PCR with electrophoresis. Results: In the YMHSB study, the mRNA expression of GM-CSF and IL-8 is significantly inhibited compared to that of control group. But the mRNA expression of IL-6 is not significantly inhibited. In the RCM study, the mRNA expression of GM-CSF and IL-S is significantly inhibited compared to that of control group. But the mRNA expression of IL-6 is not significantly inhibited. Conclusions: This study shows that YMHSB and RCM have dose-dependent inhibitory effects on the mRNA expression of IL-S and GMCSF in human epithelial cells. So these herbal medicines may inhibit the inflammatory process of asthma. Advanced studies are required to investigate the mechanisms of inhibition by herbal medicine in the asthma model.
Journal of Physiology & Pathology in Korean Medicine
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v.20
no.1
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pp.69-75
/
2006
In the present study, the author tried to investigate whether four oriental medical prescriptions named, jawan-chihyosan (CHS), gwaru-jisiltang (GJT), and several single compounds, kaempferol, coumarin, betaine and ursolic acid significantly affect mucin release from cultured hamster tracheal surface epithelial (HTSE) cells. Confluent HTSE cells were metabolically radiolabeled with 3H-glucosamine for 24 hrs and chased for 30 min in the presence of CHS, GJT, kaempferol, coumarin, betaine and ursolic acid, respectively, to assess the effect of each agent on 3H-mucin release. Possible cytotoxicities of each agent were assessed by measuring lactate dehydrogenase(LDH) release. Additionally, total elution profiles of control spent media and treatment sample (CHS and GJT) through Sepharose CL-4B column were analysed and effect of CHS and GJT on MUC5AC mRNA expression in cultured HTSE cells were investigated. The results were as follows : (1) CHS and GJT significantly stimulated mucin release from cultured HTSE cells, with significant cytotoxicity , (2) CHS and GJT chiefly stimulated the 'mucin' release and did not affect significantly the release of the other releasable glycoproteins with less molecular weight than mucin. This result suggests that the three herbal prescriptions specifically stimulate the release of mucin ; (3) CHS and GJT significantly increased the expression levels of MUC 5AC mRNA. This result suggests that the three herbal prescriptions can affect the synthesis of mucin at gene level in cultured HTSE cells ; (4) Kaempferol and coumarin did not affect mucin release, however, betaine and ursolic acid stimulated mucin release. All the agents did not show significant cytotoxicity. We suggest that the effects of CHS and GJT, betaine and ursolic acid should be further investigated and it is of great value to find, from oriental medical prescriptions, novel agents which have the effective expectorant or mucoregulative effect on mucin secretion from airway goblet cells.
Kim, Sook-Hee;Kim, Jin-Sook;Jin, Mi-Rim;Kim, Ha-Won;Choi, Eung-Chil;Kim, Byong-Kak
Korean Journal of Pharmacognosy
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v.24
no.4
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pp.267-281
/
1993
To find antitumor components from higher fungi, the mycelia of Collybia confluens (Pers. ex Fr.) Kummer were cultured in artificial media. For efficient production of the mycelia, the influences of various modifications of culture conditions were examined. A water-soluble protein-bound polysaccharide fraction, Fr. A, was obtained from the mycelia by hot water extraction. When Fr. A was purified and fractionated by DEAE-cellulose and Sepbadex G-200 gel filtration chromatographies into four fractions which were designated B, C, C-I and C-II. The tumor inhibition ratios of these fractions ranged from 46% to 75% against the solid forms of sarcoma 180 in ICR mice at doses of 20 and 50 mg/kg/day when given intraperitoneally. Especially, Fr. C which was named Collyban(CB) exhibited a marked life-prolonging effect of the mice against ascitic forms of sarcoma 180 at a dose of 50 mg via i.p. administration. To extend spectra of the antitumor activities and eliminate the effects of allograft rejection, the characterization of antitumor effects of CB was performed in syngeneic host-tumor systems. It did not show any antitumor activity against L1210 murine leukemia in $CD_2Fl$ mice but prolonged their life span against ascitic forms of $MM_{46}$ carcinoma in $C_3H/He$ mice. Also it exhibited antitumor activity against human cervical cancer HeLa cells that were xenografted into nude mice having BALB/c genetic backgrounds by the i.p. injection at a dose of 100 mg/kg/day. In order to characterize the antitumor components, CB was examined by chemical analysis. It was acidic protein-bound polysaccharides composed of 31% polysaccharide, 27% protein and 3% hexosamine. CB was fractionated into two fractions, Fr. C-I(M.W.: 500 Kd) and Fr. C-II(M.W.:30 and 8 Kd) by Sephadex G-200 gel filtration chromatography.
Background: Interferon lambda receptor 1 (IFNLR1) is a type II cytokine receptor that clings to interleukins IL-28A, IL29B, and IL-29 referred to as type III IFNs (IFN-λs). IFN-λs act through the JAK-STAT signaling pathway to exert antiviral effects related to preventing and curing an infection. Although the immune function of IFN-λs in virus invasion has been described, the molecular mechanism of IFNLR1 in that process is unclear. Objectives: The purpose of this study was to elucidate the role of IFNLR1 in the pathogenesis and treatment of porcine reproductive and respiratory syndrome virus (PRRSV). Methods: The effects of IFNLR1 on the proliferation of porcine alveolar macrophages (PAMs) during PRRSV infection were investigated using interference and overexpression methods. Results: In this study, the expressions of the IFNLR1 gene in the liver, large intestine, small intestine, kidney, and lung tissues of Dapulian pigs were significantly higher than those in Landrace pigs. It was determined that porcine IFNLR1 overexpression suppresses PRRSV replication. The qRT-PCR results revealed that overexpression of IFNLR1 upregulated antiviral and IFN-stimulated genes. IFNLR1 overexpression inhibits the proliferation of PAMs and upregulation of p-STAT1. By contrast, knockdown of IFNLR1 expression promotes PAMs proliferation. The G0/G1 phase proportion in IFNLR1-overexpressing cells increased, and the opposite change was observed in IFNLR1-underexpressing cells. After inhibition of the JAK/STAT signaling pathway, the G2/M phase proportion in the IFNLR1-overexpressing cells showed a significant increasing trend. In conclusion, overexpression of IFNLR1 induces activation of the JAK/STAT pathway, thereby inhibiting the proliferation of PAMs infected with PRRSV. Conclusion: Expression of the IFNLR1 gene has an important regulatory role in PRRSV-infected PAMs, indicating it has potential as a molecular target in developing a new strategy for the treatment of PRRSV.
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