• Journal of Microbiology and Biotechnology
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• v.27 no.6
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• pp.1090-1097
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• 2017

Rhus verniciflua Stokes (RVS), an herbal medicine found in East Asia, was extracted and further fractionated to investigate its antioxidant capacity and neuroprotective effects. The RVS ethyl acetate (EtOAc) fraction had the highest level of total phenolics and antioxidant capacity among all solvent fractions tested. Pretreatment of PC-12 cells with the EtOAc fraction effectively attenuated $H_2O_2$-induced oxidative damage. Furthermore, the EtOAc fraction significantly attenuated caspase-3 activity, resulting in inhibition of $H_2O_2$-induced apoptosis. We identified and quantified fustin, sulfuretin, and butein in the EtOAc fraction using accurate mass quadrupole time-of-flight mass spectrometry and reversed-phase high-performance liquid chromatography. The intracellular antioxidant capacity and superoxide dismutase (SOD) activity were significantly increased in PC-12 cells treated with the EtOAc fraction and with individual flavonoids. When cells were pretreated with the EtOAc fraction or individual flavonoids and then co-incubated with diethyldithiocarbamic acid (an inhibitor of SOD activity), cell viability against $H_2O_2$-induced oxidative stress was attenuated. These results suggest that the RVS EtOAc fraction and its flavonoid constituents protect PC-12 cells against $H_2O_2$-induced neurotoxicity through their antioxidant properties.

• Journal of Physiology & Pathology in Korean Medicine
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• v.17 no.1
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• pp.203-208
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• 2003

Antioxidant effects of Rehmannia vaporata, Discorea Radix, Corni Fructus, Hoolen, Alismatis Radix, and Mountain Cortex Radicis composing Yukmijihwang-tang were studied. The results are as follows; 1. As a result of detecting the defensive effect of each component on cell damage, only the survival rate of cells with 10 ㎎/㎖ Mountain Cortex Radicis was significantly increased. 2. Next, we examined the inhibitory effects of them on ROS occurrence. The result showed significant inhibition of ROS occurrence in cells; with 10 ㎎/㎖ Rehmannia vaporata, cells with 10 ㎎/㎖ Corni Fructus, and cells with 10 ㎎/㎖ Mountain Cortex Radicis. Since the cells with 10 ㎎/㎖ Rehmannia vaporata, however, showed significant cypotoxicity, its result is not meaningful. 3. Finally, the investigation of ROS occurrence / cell found that Corni Fructus and Mountain Cortex Radicis had significant inhibitory effect on ROS occurrence.

• The Journal of Korean Medicine
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• v.21 no.4
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• pp.64-72
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• 2000

Objectives : This study was designed to investigate the neuroprotective effect of Hwansodan(Huanshao-dan) on the apoptosis induced by withdrawal of neurotrophic support. Methods : PCl2 pheochromocytoma cells have been used extensively as a model for studying the cellular and molecular effects of neuronal cells. The viability of cells was measured by MTT assay. We used DNA fragmentation and caspase 3-like protease activation assay. Results : The water extract of Hwansodan(Huanshao-dan) significantly showed protective effects on serum and glucose deprivation-induced apoptotic death. Hwansodan(Huanshao-dan) also prevents DNA fragmentation and caspase 3-like protease activation, representing typical neuronal apoptotic phenomena in PCl2 pheochromocytoma cells and induces tyrosine phosphorylation of proteins around 44 kDa, which was identified as ERK1 with electrophoretic gel mobility shift by Western blot. In addition, MAPK kinase(MEK) inhibitor PD98059 and Ras inactivator, ${\alpha}-hydroxyfarnesylphosphonic$ acid attenuated the neuroprotective effects of Hwansodan(Huanshao-dan) in serum-deprived PCl2 cells. Conclusions : These results indicate that Ras/MEK/ERK signaling pathway plays a key role in neuroprotective effects of Hwansodan(Huanshao-dan) in serum-deprived PCl2 cells. Taken together, we suggest the possibility that Hwansodan(Huanshao-dan) might provide a neurotrophic-like activity in PCl2 cells.

• Proceedings of the PSK Conference
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• 2002.10a
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• pp.416.3-417
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• 2002

Among the promising cancer therapy is targeting of the drug to tumor cells via receptor specific ligands. CR2945, $\beta$-2-( ［2-(8-azaspiro ［4.5］ dec-8-ylcarbony!)-4.6-dimethylphenyl］amino-2-oxoethyl］ -(R)-1-naphthalenepropanoic acid. is known to have an inhibitory effect on a gastrin receptor of colorectal cancer cells. As the human pancreatic cancer cells (BxPC-3) express gastrin receptors. interruption of binding of gastrin with gastrin receptor of human pancreatic cancer cells by CR2945 inhibits the growth of human pancreatic cancer cells. (omitted)

• Animal cells and systems
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• v.6 no.2
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• pp.167-170
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• 2002

The long terminal repeats (LTRs) of human endogenous retrovirus (HERV) have been found to be coexpressed with sequences of closely located genes. It has been suggested that the LTR elements have contributed to the structural change or genetic variation of human genome connected to various diseases and evolution. We examined the HERV-W LTR elements in various cancer cells (2F7, A43l , A549, HepG2, MIA-PaCa-2, PC-3, RT4, SiHa, U-937, and UO-31). Using genomic DNA from the cancer cells, we performed PCR amplification and identified twelve new HERV-W LTR elements. Those LTR elements showed a high degree of sequence similarity (88-99％) with HERV-W LTR (AF072500). A phylogenetic tree obtained by the neighbor-joining method revealed that HERV-W LTR elements could be mainly divided into two groups through evolutionary divergence. Three HERV-W LTR elements (RT4-2, A43l-1, and UO3l-2) belonged to Group 1, whereas nine LTR elements (2F7-2, A549-1, A549-3, HepG2-3, MP2-2, PC3-1, SiHa-8, SiHa-10, and U937-1) belonged to Group 11. Taken together, our new sequence data of the HERV-W LTR elements may contribute to an understanding of tissue-specific cancer by genomic instability of LTR integration.

• Proceedings of the PSK Conference
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• 2003.10b
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• pp.69.3-70
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• 2003

A substantial body of evidence indicates that reactive oxygen intermediates (ROIs) are implicated in pathogenesis of diverse human diseases, including cancer, diabetes and neurodegenerative disorders. Oxidative stress induced by ROIs often causes cell death via apoptosis that is regulated by a plenty of functional genes and their protein products. In the present work, we have investigated the role of bcl-2 in protecting against oxidative death induced by hydrogen peroxide in cultured rat pheochromocytoma (PC12) cells. (omitted)

• Proceedings of the PSK Conference
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• 2003.10b
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• pp.79.2-79.2
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• 2003

Minocycline is known to protect neurons from microglia-mediated cell death in many experimental models of brain diseases including ischemic stroke, Huntington's disease (HD), amyotrophic lateral sclerosis (ALS), traumatic brain injury, multiple sclerosis, and Parkinson's disease. Activation of caspase-2, 3, 8, and 9 was evident within 2-8 hr following oxidative insult with 0.5 mM hydrogen peroxide in PC12 cells. Minocycline significantly attenuated activation of these caspases up to 18 hr, resulting a significant increase in cell viability as assessed by MTT assay. (omitted)

• Biomolecules & Therapeutics
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• v.22 no.4
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• pp.355-362
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• 2014

We have developed a fully automated high throughput drug screening (HTDS) system based on the microfluidic cell culture array to perform combinational chemotherapy. This system has 64 individually addressable cell culture chambers where the sequential combinatorial concentrations of two different drugs can be generated by two microfluidic diffusive mixers. Each diffusive mixer has two integrated micropumps connected to the media and the drug reservoirs respectively for generating the desired combination without the need for any extra equipment to perfuse the solution such as syringe pumps. The cell array is periodically exposed to the drug combination with the programmed LabVIEW system during a couple of days without extra handling after seeding the cells into the microfluidic device and also, this device does not require the continuous generation of solutions compared to the previous systems. Therefore, the total amount of drug being consumed per experiment is less than a few hundred micro liters in each reservoir. The utility of this system is demonstrated through investigating the viability of the prostate cancer PC3 cell line with the combinational treatments of curcumin and tumor necrosis factor-alpha related apoptosis inducing ligand (TRAIL). Our results suggest that the system can be used for screening and optimizing drug combination with a small amount of reagent for combinatorial chemotherapy against cancer cells.

• BMB Reports
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• v.39 no.6
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• pp.743-748
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• 2006

The complement (C) regulatory proteins decay accelerating factor (DAF, CD55) and CD59 could protect host cells using different mechanisms from C-mediated damage at two distinct levels within the C pathway. Co-expression of DAF and CD59 would be an effective strategy to help overcome host C-induced xenograft hyperacute rejection. In this study, we made a construct of recombinant expression vector containing DAF and CD59 cDNA and the stable cell lines were obtained by G418 selection. Extraneous genes integration and co-expression were identified by PCR, RT-PCR and Western blot analysis. Human c-mediated cytolysis assays showed that NIH/3T3 cells transfected stably with pcDNA3-CD59, pcDNA3-DAF, and pcDNA3-CD59DAF-DP were protected from C-mediated damage and that synchronously expressed human CD59 and DAF provided the most excellent protection for host cells as compared with either human CD59 or DAF expressed alone. Therefore, the construct represents an effective and efficacy strategy to overcome C-mediated damage in cells and, ultimately, in animals.

• The Journal of Internal Korean Medicine
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• v.33 no.2
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• pp.197-208
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• 2012

Objectives : Gleditsiae Spina has the effects of expelling toxins, draining pus, invigorating blood and resolving abscesses. Some clinicians apply the herb for patients suffering from cancer. However, its anti-cancer activities are not well understood. In the present study, anti-tumor agents from Gleditsiae Spina were purified. Methods : The viability of the PC-3 cell line was determined using MTT assay, and the induction of apoptosis by Gleditsiae Spina extract in PC-3 cells was measured by Annexin-V/propidium iodide double staining assay detected by flow cytometry. TLC and HPLC analysis were used to separate and identify the anti-cancer agents. Results : Treatment of the extract resulted in significant decreased cell viability of PC-3 cells in a dose- and time-dependent manner. Dose-dependent apoptotic cell death was also measured by flow cytometry analysis. The anti-cancer agents were successfully separated and identified by using TLC and HPLC analysis and the most potential agent among them was separated from EtOAC fraction. Conclusions : These results might be applied in developing new drugs from natural resources like Korean traditional medicine, and also support the clinical usefulness of herbal medicine.