• Food Science and Biotechnology
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• v.18 no.1
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• pp.84-88
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• 2009

Cathepsin S is a cysteine protease that affects extracellular matrix remodeling. Recently, several studies have reported that cathepsin S is involved in obesity. Both mouse and human adipose cells produce this enzyme in the early phase of adipocyte differentiation, where it degrades fibronectin. Cathepsin S gene expression is elevated in the adipose tissue of obese mice as compared to that of lean mice. Paecilomyces tenuipes water extracts (PTW) are shown to have an inhibitory effect on cathepsin S activity. In this study, Z-Val-Val-Arg-MCA was used as a cathepsin S-specific substrate in order to examine inhibitory effect of PTW. Supplementing 3T3-L1 cell media with PTW clearly reduced lipid droplet accumulation and cathepsin S-induced adipogenesis. Furthermore, PTW decreased weight gain, subcutaneous adipose tissue growth, the level of serum triglyceride, and total cholesterol in mice fed a high-fat diet. These data suggest that PTW work against adipose cathepsin S and presumably contribute to anti-obese activities.

• IMMUNE NETWORK
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• v.16 no.1
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• pp.75-84
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• 2016

Cancer is one of the leading causes of morbidity and mortality worldwide; therefore there is a need to discover new therapeutic modules with improved efficacy and safety. Immune-(cell) therapy is a promising therapeutic strategy for the treatment of intractable cancers. The effectiveness of certain chemotherapeutics in inducing immunogenic tumor cell death thus promoting cancer eradication has been reported. Ginsenoside Rg3 is a ginseng saponin that has antitumor and immunomodulatory activity. In this study, we treated tumor cells with Rg3 to verify the significance of inducing immunogenic tumor cell death in antitumor therapy, especially in DC-based immunotherapy. Rg3 killed the both immunogenic (B16F10 melanoma cells) and non-immunogenic (LLC: Lewis Lung Carcinoma cells) tumor cells by inducing apoptosis. Surface expression of immunogenic death markers including calreticulin and heat shock proteins and the transcription of relevant genes were increased in the Rg3-dying tumor. Increased calreticulin expression was directly related to the uptake of dying tumor cells by dendritic cells (DCs): the proportion of CRT⁺CD11c⁺cells was increased in the Rg3-treated group. Interestingly, tumor cells dying by immunogenic cell death secreted IFN-γ, an effector molecule for antitumor activity in T cells. Along with the Rg3-induced suppression of pro-angiogenic (TNF-α) and immunosuppressive cytokine (TGF-β) secretion, IFN-γ production from the Rg3-treated tumor cells may also indicate Rg3 as an effective anticancer immunotherapeutic strategy. The data clearly suggests that Rg3-induced immunogenic tumor cell death due its cytotoxic effect and its ability to induce DC function. This indicates that Rg3 may be an effective immunotherapeutic strategy.

• Journal of Microbiology and Biotechnology
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• v.33 no.2
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• pp.211-218
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• 2023

A cryptic plasmid (pTH32) was characterized from Tetragenococcus halophilus 32, an isolate from jeotgal, Korean traditional fermented seafood. pTH32 is 3,198 bp in size with G+C content of 35.84%, and contains 4 open reading frames (ORFs). orf1 and orf2 are 456 bp and 273 bp in size, respectively, and their translation products showed 65.16% and 69.35% similarities with RepB family plasmid replication initiators, respectively, suggesting the rolling-circle replication (RCR) mode of pTH32. orf3 and orf4 encodes putative hypothetical protein of 186 and 76 amino acids, respectively. A novel Tetragenococcus-Escherichia coli shuttle vector, pMJ32E (7.3 kb, Em^r), was constructed by ligation of pTH32 with pBluescript II KS(+) and an erythromycin resistance gene (ErmC). pMJ32E successfully replicated in Enterococcus faecalis 29212 and T. halophilus 31 but not in other LAB species. A pepA gene, encoding aminopeptidase A (PepA) from T. halophilus CY54, was successfully expressed in T. halophilus 31 using pMJ32E. The transformant (TF) showed higher PepA activity (49.8 U/mg protein) than T. halophilus 31 cell (control). When T. halophilus 31 TF was subculturd in MRS broth without antibiotic at 48 h intervals, 53.8% of cells retained pMJ32E after 96 h, and only 2.4% of cells retained pMJ32E after 14 days, supporting the RCR mode of pTH32. pMJ32E could be useful for the genetic engineering of Tetragenococcus and Enterococcus species.

• THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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• v.3 no.1
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• pp.67-84
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• 1997

Bujeonghangamtang(扶正抗癌湯) has been used for cure of tumor as a traditional medicine without any experimental evidence to support the rational basis for its clinical use. This study was carried out to evaluate the possible therapeutic or antitumoral effects of Bujeonghangamtang extract against tumor, and to carry out some mechanisms responsible for its effect. Some kinds of tumor were induced by the typical application of 3-methylcholanthrene. (MCA) or by the implantation(s.c) of malignant tumor cells such as leukemia cells(3LL cells) or sarcoma cells(Sl80 cells). Treatment of the Bujeonghangamtang water-extract (dailly 1mg／mouse, i. p.) was continued for 7 days prior to tumor induction and after that the treatment was lasted for 20 days. Against squamous cell carcinoma induced by MCA, Bujeonghangamtang decreased not only the frequency of tumor production but also the number and the weight of tumors per tumor bearing mice (TBM). Bujeonghangamtang also significantly suppressed the development of 3LL cell and S180 cell-implanted tumors in occurrence-frequency and their size, and some developed tumors were regressed by the continuous treatment of Bujeonghangamtang extract into TBM. In vitro, treatment of Bujeonghangamtang extract had no effect on the growth of some kinds of cell line such as FsaII, A431 strain but significantly inhibited the proliferation of 3LL, S180 cells and augmented the DNA synthesis of mitogen-activated lymphocytes. Bujeonghangamtang also stimulated the migrative ability of leukocyte, the MIF and IL-2 production of T lymphocytes, but not IL 6 production of B cells. Bujeonghangamtang-administration to mice enhanced NK cells activities. These results demonstrated that Bujeonghangamtang extract exhibited a significant prophylactic benefits against tumors and its antitumor activity was manifested depending on the type of tumor cells. And these results also suggested that effect of Bujeonghangamtang might be chiefly due to nonspecific enhancement of NK cell activities and cell-mediated immune responses.

• The Journal of Internal Korean Medicine
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• v.30 no.1
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• pp.74-84
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• 2009

Objectives : This study was performed to investigate the anti-fibrogenic effect of Injinchunggan-tang on cultured rat hepatic stellate cells. Materials and Methods : Hepatic stellate cells(HSC-T6) were treated with various concentrations of Injinchunggan-tang extract for 24, 48, and 72 hours. The extraction was done with distilled water. After the treatment, cell viability, proliferation, procollagen levels and the mRNA of the TIMP-1, TIMP-2, and ASMA were measured by using MTT assay. BrdU assay, procollagen type I C-peptide EIA kit and RT-PCR. Results : The proliferation, mRNA expression and synthesis of collagen of the hepatic stellate cells were inhibited by Injinchunggan-tang treatment in a dose-dependent manner. This indicates the prescription has inhibitory effect on fibrogenesis of the liver by regulating the fibrogenesis associated genes in transcription. Cell viability was inhibited in time- and dose-dependent manners. It seemed that the drug should be used with sufficient dose to acquire treatment effect. Conclusion : These results suggest that Injinchunggan-tang is beneficial in the treatment of cirrhotic patients as well as for the patients with chronic hepatitis.

• Transactions of the Korean Society of Mechanical Engineers A
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• v.28 no.8 s.227
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• pp.1183-1189
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• 2004

A novel cell count method, which can improve the count efficiency and reduce contamination problem, was presented using micro lattice engraved on culture dish. The micro lattice has feature of $50{\mu}m{\times}50{\mu}m$ rectangular shape, $2{\mu}m$ line width, and $2{\mu}m$ depth in $3mm{\times}3mm$ area. In this paper, nickel mold was fabricated with thickness of 3mm and diameter of 80 mm, and transcription of the micro lattice on a polystyrene cell culture dish was performed by hot embossing at $200\;^{\circ}C$. The tedious and error-prone harvest/load processes of conventional cell counts with a hemocytometer could be omitted, and these advantages became magnified during periodical counts involving long-term cultures. SupT1 cells and HeLa cells were cultivated with the dish for 7 days in $CO_2$ incubator and counted as $371.84/mm^2$ and $123.36/mm^2$, respectively, during the cultivation without harmful effects on the cells.

• The Journal of Korean Obstetrics and Gynecology
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• v.20 no.1
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• pp.32-49
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• 2007

Purpose: Breast cancer is the most common disease in Korean women. Despite remarkable improvements in treatment strategies against various cancer during the past 40 years, breast cancer still remains as one of the main causes of cancer mortality among women in the whole world. This study was carried out to investigate antioxidative and anti-proliferative effects on MCF-7 human breast cancer cells of Ikiyangyoung-Tang extract. Methods: We measured a content of polyphenol and flavonoid in the Ikiyangyoung-Tang extract, eliminative ability of DPPH radical, ABTS free radical and hydrogen peroxide, antioxidative effects of linoleic acid, cytotoxicity on MCF-7 human breast cancer cells. MCF-7 cells were cultured in Dulbecco's modified Eagle's medium/F12(DMEM/F12) supplemented with 10 % fetal bovine serum(FBS; Gibco) and antibiotics. Results : The extract of Ikiyangyoung-Tang contains polyphenol of 168.3${\pm}$12.8 ${\mu}$g/mg and flavonoid of 84.3${\pm}$3.4 ${\mu}$g/mg. Above results show profitable abilities of elimination of ${\alpha}$-${\alpha}$-Diphenyl-${\beta}$-picrylhydrazyl(DPPH) radical, ABTS free radical and hydrogen peroxide. Also, the extract of Ikiyangyoung-Tang strongly inhibits the proliferation of MCF-7 cells in a dose ependent manner. And. it has cytotoxicity on NIH3T3 cells. Conclusion : It can be concluded that Ikiyangyoung-Tang extract has an antioxidative effect and antiproliferative effect on MCF-7 human breast cancer cells.

• Journal of the Korean Society of Food Science and Nutrition
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• v.40 no.11
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• pp.1525-1531
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• 2011

Antioxidant activity and protective effects of extracts from Helianthus tuberosus L. leaves (HTL) on t-BHP-induced oxidative stress in human liver Chang cells were investigated. The total polyphenol and flavonoid content of the water and ethanolic extracts from HTL were 89.6${\pm}$1.96, 94${\pm}$2.03 mg gallic acid equivalent/g extract, and 65.1${\pm}$2.84, 54.6${\pm}$1.87 mg catechin equivalent/g extract, respectively. In addition, $IC_{50}$ values for 1,1-diphenyl-2-picrydrazyl (DPPH), alkyl, and hydroxyl radical scavenging activity of the water extracts were 0.010${\pm}$0.003 mg/mL, 0.014${\pm}$0.002 mg/mL, and 0.989${\pm}$0.003 mg/mL, respectively. Antioxidant activities of the extracts were also determined by ferric reducing antioxidant power (FRAP), 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging activity and reducing power. The HTL extracts showed a strongly inhibitory effect on lipid peroxidation by measuring ferric thiocyanate (FTC) and thiobarbituric acid (TBA) values. In an MTT assay on the Chang cells, the extracts showed a protective effect by increasing cell viability and decreasing ROS on t-BHP-induced oxidative stress in Chang cells. These results indicate that the HTL extracts possess an antioxidant activity.

• The Korean Journal of Physiology and Pharmacology
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• v.11 no.6
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• pp.247-251
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• 2007

Changes of single unit activity of CA1 hippocampus region were investigated in anesthetized Mongolian gerbils for six days following transient ischemia. Ischemia was produced immediately before the implantation of micro-wire recording electrodes. In control animals receiving pseudo-ischemic surgery, neither spontaneous neuronal activities ($5.70{\pm}0.4Hz$) nor the number of recorded neurons per animal changed significantly for six days. Correlative firings among simultaneously recorded neurons were weak (correlation coefficient > 0.6) in the control animals. Animals subjected to ischemia exhibited a significant elevation of neural firing at post-ischemic 12 hr ($9.95{\pm}0.9Hz$) and day 1 ($8.48{\pm}0.8Hz$), but a significant depression of activity at post-ischemic day 6 ($1.84{\pm}0.3Hz$) when compared to the activities of non-ischemic control animal. Ischemia significantly (correlation coefficient > 0.6) increased correlative firings among simultaneously recorded neurons, which were prominent especially during post-ischemic days 1, 2 and 6. Although the numbers of spontaneously active neurons recorded from control group varied within normal range during the experimental period, those from ischemic group changed in post-ischemic time-dependent manner. Temporal changes of the number of cells recorded per animal between control group and ischemic group were also significantly different (p = 0.0084, t = 3.271, df = 10). Cresyl violet staining indicated significant loss of CA1 cells at post-ischemic day 7. Overall, we showed post-ischemic time-dependent, differential changes of three characteristics, including spontaneous activity, network relationship and excitability of CA1 cells, suggesting sustained neural functions. Thus, histological observation of CA1 cell death till post-ischemic day 7 may not represent actual neuronal death.

• Reproductive and Developmental Biology
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• v.35 no.3
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• pp.197-201
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• 2011

This study was conducted to investigate the effects of donor cell treatments on the production of transgenic cloned piglets. Ear fibroblast cell obtained from NIH MHC Inbred minipig was used as control. The GalT knock-out/CD45 knock-in (GalT/CD46) transgenic cell lines were established and used as donor cells. The reconstructed GalT/CD46 embryos were surgically transferred into oviduct of naturally cycling surrogate sows (Landrace ${\times}$ Yorkshire) on the second day of standing estrus. Unlike control (1.2 kV/cm, 75.4%), the fusion rate of the GalT/CDl6 donor cells was significantly higher in 1.5 kV/cm, (84.5%) than that of 1.25 kV/cm, (20.2%) (p<0.01). When the number of the transferred embryos were more than 129, the pregnancy and delivery rates were increased to 13/20 (65%) and 5/20 (25%) compared to less then 100 group [1/6 (16.7%) and 0/6 (0%)], respectively. To analyze the effect of donor cell culture condition on pregnancy and delivery rates, the GalT/CD46 donor cells were cultured with DMEM or serum reduced medium. In serum reduced medium group, the pregnancy and delivery rates were improved to 8/12 (66.7%) and 5/12 (41.7%) compared to DMEM group [3/7 (42.9%) and 0/7 (0%)], respectively. In conclusion, it can be postulated that an appropriate fusion condition and culture system is essential factors to increase the efficiency of the production of transgenic cloned piglets.