• Clinical and Experimental Reproductive Medicine
• /
• 제41권1호
• /
• pp.1-8
• /
• 2014

Objective: Estrogen related receptor ${\beta}$ (Esrrb) is a member of the orphan nuclear receptors and may regulate the expression of pluripotencyrelated genes, such as Oct4 and Nanog. Therefore, in the present study, we have developed a method for delivering exogenous ESRRB recombinant protein into embryos by using cell-penetrating peptide (CPP) conjugation and have analyzed their effect on embryonic development. Methods: Mouse oocytes and embryos were obtained from superovulated mice. The expression of Oct4 mRNA and the cell number of inner cell mass (ICM) in the in vitro-derived and in vivo-derived blastocysts were first analyzed by real time-reverse transcription-polymerase chain reaction and differential staining. Then 8-cell embryos were cultured in KSOM media with or without $2{\mu}g/mL$ CPP-ESRRB protein for 24 to 48 hours, followed by checking their integration into embryos during in vitro culture by Western blot and immunocytochemistry. Results: Expression of Oct4 and the cell number of ICM were lower in the in vitro-derived blastocysts than in the in vivo-derived ones (p<0.05). In the blastocysts derived from the CPP-ESRRB-treated group, expression of Oct4 was greater than in the non-treated groups (p<0.05). Although no difference in embryonic development was observed between the treated and non-treated groups, the cell number of ICM was greater in the CPP-ESRRB-treated group. Conclusion: Treatment of CPP-ESRRB during cultivation could increase embryos' expression of Oct4 and the formation rate of the ICM in the blastocyst. Additionally, an exogenous delivery system of CPP-conjugated protein would be a useful tool for improving embryo culture systems.

• 한국가축번식학회지
• /
• 제23권3호
• /
• pp.247-256
• /
• 1999

본 연구는 한국야생 유래 행동이상 마우스(M. m. molossinus-tt@Kist)의 혈액학 및 혈액생화학치와, 체외수정란과 정자동결보존법, 체외수정과 수정란 이식기법을 이용한 번식장애 개선과 병원성 미생물이 제거된 무병마우스 생산을 위하여 실시하였다. 1. 5주령의 혈액학치에서 RBC, platelet치는 근교계에 비해 높게 나타났다. 혈액생화학치에서는 total cholesterol치가 근교계에 비해 높게 나타났으나 triglyceride, total protein, albumin치는 유사하였다. 2. 과배란 유기시의 평균 채란 수는 PMSG/hCG를 2.5/2.5 lU 투여군에서 11.6개, 5.0/5.0 IU 투여군에서 12.7개로 통계학적 유의 차는 인정되지 않았다. 3. 2.5/2.5 lU 투여군과 5.0/5.0 lU 투여군의 수정율은 각각 87.9%와 52%로 2.5/2.5 lU 부여군이 유의성있게 높은 성적을 나타내었고(p<0.05), 2세포기로의 발달율도 각각 유의성이 인정되는 99.0%와 90.6%였다 (p<0.05). 4. 동결정자를 이용한 체외수정에서의 수정율은 24.8%로 신선정자를 사용했을 때의 87.9% 보다 낮은 성적이었다. 5. 체외 수정란의 이식후의 산자율은 동결 2세포기 체외수정란의 경우 5마리(6.6%), 동결 정사를 이용한 체외 수정란의 경우 6마리(19.4%)와 대조군의 체외수정란의 경우 10마리(21.7%)의 새끼를 얻었다. 6. 이식후 출산한 산자의 미생물학적 검사에서 MHV(Mouse hepatitis virus)와 Staphylo-coccus aureus 등의 병원성 미생물이 무병대리모를 이용한 수정란이식에 의해 제거되었음을 확인하였다.

• 한국산림과학회지
• /
• 제87권2호
• /
• pp.260-269
• /
• 1998

본 연구는 전통적인 민간요법으로 많이 이용되고 있는 옻나무의 옻칠액의 주성분인 urushiol의 암세포증식억제 효과를 검정하기 위하여 L1210(mouse 혈액암세포), PC-9(인체폐암세포), A427(인체폐암세포), KATO III(인체위암세포)에 대한 urushiol의 세포독성을 검정하였다. Urushiol은 column chromatography에서 hexane과 acetone을 혼합하여 step-wise법으로 용출한 분획을 암세포주 L1210, PC-9, A427, KATO III에 투여했다. 그 결과 암세포 증식억제효과는 hexane : acetone(9 : 1) 분획이 가장 높았고, 그외 50%억제농도($IC_{50}$)는 A427 에서 $0.018{\mu}g/m{\ell}$로 가장 우수하였다. 이들 암세포에 대한 urushiol의 hexane: acetone(9 : 1)분획의 50%억제농도($IC_{50}$)를 시판용 tetraplatin 비교한 결과 urushiol이 tetraplatin에 대하여 L1210에서 3.4배, PC-9 에서 3.9배, A427에서 105.5배의 낮은 농도에서도 동일한 억제효과를 나타냈고, KATO III에서는 다소 높았다.

• Archives of Pharmacal Research
• /
• 제17권3호
• /
• pp.135-138
• /
• 1994

Acyclonucleoside homologues of 1-$(\omega$-cyabnoalkyl)-and 1, 3-bis $(\omega$-cyanoalkyl) uracils were synthesized by the series of alkylation reactions of uracil with the $\omega$-chloroalkyl nitrile ${(Cl-(CH}_2)_n$-CN;n=1, 2, 3, 4) in DMSO under $50-70^circ{C}$ temperature. The 1-$(\omega$-cyanoalkyl)-and 1, 3-bis$(\omega$-cyanoalkyl) uracils were separated either by the fractional crystallization or column chromatography. The antitumor activities for these synthesized compounds were determined against four cell lines (J-82 cell, P388 cell, FM-3A cell and U-937 cell lines). These compounds failed to exhibit any significant antitumor activity.

• 동의생리병리학회지
• /
• 제26권1호
• /
• pp.26-34
• /
• 2012

This study is to investigate the effects of Chinemys reevesii (CR) on allergic inflammation mechanism related chronic dermatitis. To investigate the effects of CR, we study inhibitory effect of CR on the levels of pro-inflammatory cytokines released from RAW 264.7 cell stimulated with lipopolysaccaride (LPS), and EoL-1, THP-1, Jutkat cell stimulated with Dermatophagoides pteronyssinus (DP), and LPS induced acute inflammatory BALB/c mouse model. CR reduced the levels of IL-$1{\beta}$ released from RAW 264.7 cell stimulated with LPS at 20 ug/ml, 10 ug/ml concentration. CR significantly reduced the levels of MCP-1 released from EoL-1 cell, IL-6 from THP-1 cell, and IL-4, IL-5, TNF-${\alpha}$ from Jutkat cell stimulated with DP at all the concentration. CR significantly reduced the levels of TNF-${\alpha}$, IL-6, IL-$1{\beta}$, in LPS induced inflammatory BALB/c mouse model, in a dose-dependent manner. These results suggested that CR has suppressive effects on pro-inflammatory cytokines in various inflammation related cell lines through the regulation of immune system. CR could be a therapeutic agent for treatment of chronic inflammatory dermatitis in the future.

• Asian Pacific Journal of Cancer Prevention
• /
• 제17권4호
• /
• pp.2083-2087
• /
• 2016

A new compound namely (13-(3,3-dihydroxypropyl)-1,6-dihydroxy-3,4-dihydro-1H-isochromen-8(5H)-one (1) was isolated from an ethyl acetate extract of the borne fungi Screlotium rolfsii. Its chemical structure was elucidated by spectroscopic analysis. Screlotiumol 1 were evaluated for their effects on the reversion of multidrug resistant (MDR) mediated by P-glycoprotein (P-gp) of the soil borne fungi. The multidrug resistant P-glycoprotein is a target for chemotherapeutic drugs in cancer cells. In the present study rhodamine-123 exclusion screening test on human mdr1 gene transfected mouse gene transfected L5178 and L5178Y mouse T-cell lymphoma which showed excellent MDR reversing effect in a dose dependent manner against mouse T-lymphoma cell line. Moreover, molecular docking studies of compound-1 also showed better results as compared with the standard. Therefore the preliminary results obtained from this study suggest that screlotiumol 1 could be used as a potential agent for the treatment of cancer.

• Journal of Microbiology and Biotechnology
• /
• 제13권4호
• /
• pp.537-543
• /
• 2003

The mouse interferon gene (MuIFN-${\gamma}$) was cloned and then used to transform Saccharomyces cerevisiae. Expressed MuIFN-$\{gamma}$ protein (MuIFN-${\gamma}$) was successfully secreted into culture medium due to the presence oi the signal peptide of rice amylase 1A. Two different promoters fused to MuIFN-${\gamma}$ were tested: glyceraldehyde-3-phosphate dehydrogenase (GPD) promoter and a yeast hybrid ADH2-GPD (AG) promoter consisting of alcohol dehydrogenase II (ADH2) and GPD promoter. Using the hybrid promoter, the accumulation of MuIFN-${\gamma}$transcript was the highest after the 24 h cultivation, and then gradually decreased as the cultivation proceeded. However, both cell growth and recombinant MuIFN-${\gamma}$production reached their peaks after the 4-day cultivation. It was possible to produce 6.5 mg/l of MuIFN-${\gamma}$ without any changes in cell growth. Using GPD promoter, the MuIFN-${\gamma}$ transcript accumulation and the recombinant MuIFN-${\gamma}$ production followed the same pattern as the cell growth. However. compared to that of the hybrid promoter, the production of recombinant MuIFN-${\gamma}$ was 0.2 mg/l. The secreted MuIFN-${\gamma}$ had estimated molecular masses of 21 kDa and 23 kDa, which were larger than that of the encoded size due to glycosylation. The protection assay against the viral infection indicated that the recombinant MuIFN-${\gamma}$ was bioactive.

• Journal of Applied Biological Chemistry
• /
• 제63권1호
• /
• pp.89-93
• /
• 2020

In this study, the efficacy of melanoma cell B16F10 was investigated using the Korean native plant Oplismenus undulatifolius (OU). First, the cell viability of the extract was more than 90% when treated with 15 ㎍/mL of phenolics from OU. The results showed that melanin biosynthesis and cellular tyrosinase synthesis were inhibited by treatment with α-melanocyte-stimulating hormone-stimulated mouse melanoma cell B16F10 at a concentration of 15 ㎍/mL of phenolics for cell-line efficacy. The expression of tyrosinase, tyrosinase-related protein (TRP)-1, TRP-2, and microphthalmia transcription factor (MITF) protein was confirmed by western blot to investigate the effect of phenolics from OU on melanin biosynthesis. When treated with phenolics from OU 15 ㎍/mL, tyrosinase, TRP-1, TRP-2, and MITF decreased the protein expression level. In particular, tyrosinase, TRP-1, and MITF inhibited the production amount to a level similar to that of the non-treated normal group, indicating that the effect was excellent. Therefore, phenolics from OU acts as an inhibitor of tyrosinase, TRP-1, TRP-2, and its transcription factor MITF, and participates in melanin biosynthesis mechanism. These results suggested the potential for development as a material.

• 동의생리병리학회지
• /
• 제16권6호
• /
• pp.1223-1229
• /
• 2002

The purpose of this research was to investigate the effect of Cheongsimondam-tang(CSODT) on the activity of immune cell and anti-carcinogenic effect of mouse leukemia cell. The addition of CSODT(1 ㎍/ml) enhanced the proliferation of cultured-splenocytes and thymocytes. And also administration of CSODT(500 ㎍/kg) accelerated subpopulation of splenic and thymic T lymphocytes especially CD4/sup +/-T/sub H/ cells in BALB/c mice. CSODT treatment decreased cell proliferation and increased apoptotic cell death of cultured-L1210 leukemia cells, and induced apoptosis in addition to decreased mitochondrial transmembrane potential (ΔΨm) of transplanted-L1210 cells in vivo. These results suggest that CSODT have a cellular immuno-regulatory effect and anti-cancer property action.

• Archives of Pharmacal Research
• /
• 제19권1호
• /
• pp.62-65
• /
• 1996

A homologous series of twenty, hitherto unreported, analogues of 5-halosubstituted $1, 3-Bis(\omega-cyanoalkyl)uracil$acyclic nucleosides were synthesized by the series of alkylation reactions of 5-halouracils with the corresponding chloroacetonitrile, chloropropionitrile, chlorobutyronitrile and 5-chlorovaleronitrile $(Cl-(C_ 2)_n-CN: n=l, 2, 3, 4)\; in\; anhydrous\; DMSO\; (or DMF)/K_2CO_3(or NaH)\; under\; 75^{\circ}C$ temperature. Antitumor activities for the synthesized compounds were determined against three cell lines (FM-3A cell, P-388 cell and U-938 cell lines). The compounds that exhibited moderate activity to significant activity, included la-b, 2a-b, 3a-c, and 4a, whose compounds were active against P-388, FM-3A and U-937 cell lines with the compounds la, lb, and 2a, showing significant antitumor activity (inhibitory concentrations $(IC_{50})$ ranged from 2.2 to $7.0\mug/ml$). Their strucrure-activity relationship did not show any activity differences in their effective chain length (methyl, ethyl, propyl, butyl) in 1, 3-bis(.omega.-cyanoalkyl) uracils.