• Pediatric Gastroenterology, Hepatology & Nutrition
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• 제14권2호
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• pp.148-154
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• 2011

목적: 저자들은 지역사회에서, 로타바이러스 장염에 걸린 소아들에서 질환의 중증도와 로타바이러스 VP7 유전형과 VP4 유전형의 분포에 대해 알아보고자 하였다. 방법: 2007년 12월부터 2008년 6월까지 로타바이러스 장염으로 병원에 입원한 156명의 소아들에서 대변 샘플을 수집하였다. 모든 환자들에 대한 질환 중증도는 Vesikari 점수를 이용하여 평가하였다. 로타바이러스 ds-RNA를 분리한 후, 역전사 중합효소 연쇄반응과 다중 중합효소 연쇄반응을 이용하여 cDNA를 합성하였다. 마지막으로 유전형을 확인하였다. 결과: 156명의 환자 샘플에서, VP7(G)는 147명(94.2%)에서 확인되었고 VP4(P)는 140명(89.7%)에서 확인되었다. G1 유전형(147명 중 116명; 78.9%)과 P[8] 유전형(140명 중 137명)이 각각 가장 많았다. VP7과 VP4 조합형은 138명에서 확인할 수 있었는데, G1P[8] 조합형이 111명(80.4)으로 가장 많았다. 다른 조합형들은 종류가 다양하였고 분포가 낮았다. 전체 조합형 중 9.4%가 새로운 로타바이러스 백신에 포함되지 않은 것이었다. 로타바이러스 장염의 질환 중중도는 $11.8{\pm}3.3$ ($mean{\pm}2SD$)였다. 전체 환자에서 경증 내지 중등도가 37.8%였고, 중증은 62.2%였다. 결론: 로타바이러스 유전자 조합형 중 가장 흔한 것은 G1P[8]이었다. 새로운 백신에 포함되지 않은 유전자 조합형은 9.4%였다. 로타바이러스 장염으로 입원한 소아들의 질환 중증도분포는 경증과 중등도보다 중증에서 높게 나타났다.

• 생명과학회지
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• 제14권4호
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• pp.650-656
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• 2004

야생벼의 일종인 Oryza grandiglumis (CCDD, 2n=48)는 도열병, 잎집무늬마름병, 흰빛잎마름병, 그리고 벼멸구와 같은 병충해에 저항성을 가지는 것으로 알려져 있다. 이와 같은 곰팡이와 해충에 반응하여 차별 발현하는 유전자를 클로닝 하기 위하여 상처처리와 yeast extract를 Oryza grandiglumis에 0시간과 24시간 각각 처리하였다. 유전자의 클로닝을 위하여 희귀 발현유전자의 클로닝에 효율적인 것으로 알려진 Suppression Subtractive Hybridization (SSH) 방법이 처리 후 24시간 된 식물을 재료로 사용되었다. 그 결과, 776개의 cDNA clones이 확보되었으며, 유전자 발현의 진위여부를 빠르게 스크린하기 위하여 colony array가 수행되었다. 115개의 colony가 positive로 판명되었고, 이들의 평균 insert size는 400 bp에서 700 bp에 이르렀고, 이들에 대한 염기서열 분석이 수행되었다. 염기서열 분석 결과, 68개 clone들이 알려진 기능의 유전자와 homology를 나타냈으며, 이중에서 16개 clone이 일차대사에 관련된 것과 유사성을, 5개가 plant retrotransposon과 유사성을, 5개가 식물 방어기작 관련 metallothionein-like gene과 염기서열 유사성을 보였다. 이외에 다양한 유전자들이 아미노산 합성관련, membrane transport, signal transduction등에 관여하는 유전자들과 상동성을 나타내었다. 이들 유전자중에서 4 개의 클론(ogwfi-161, ogwfi-646, ogwfi-663, ogwfi-695)들이 선발되었고 이들에 대한 Northern 분석이 수행되었다. Northern 분석 결과 ogwfi-161, ogwfi-646, ogwfi-663, ogwfi-695는 wounding과 yeast extract처리 에 의한 차별 발현이 확인되었다. 이상의 결과를 종합하여 볼 때, SSH방법은 병충해등과 같은 조건에 의해 차별 발현되는 유전자들을 빠른 시간 내에 다량으로 발굴할 수 있는 매우 효율적인 방법이라고 생각된다.

• 한국양식학회지
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• 제18권4호
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• pp.293-298
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• 2005

RT-PCR을 이용하여 감성돔의 뇌하수체에서 발현하는 FSH$\beta$와 LH$\beta$ cDNA를 각각 cloning하였다. GenBank를 이용하여 FSH$\beta$와 LH$\beta$ 아미노산 배열과 상동성을 비교한 결과, 감성돔 FSH$\beta$는 참돔 FSH$\beta$와 $83.3\%$, snakehead fish FSH$\beta$와 $79.2\%$ 및 striped bass FSH$\beta$와 $76.0\%$의 상동성이 나타났으며 , 감성돔 LH$\beta$는 yellowfin seabream LH$\beta$와 $97.7\%$, 참돔 LH$\beta$와 $96.2\%$의 높은 상동성을 보였다. 또한, 본 연구에서는 1년생 수컷 감성돔에 GnRHa를 처리한 후, 뇌하수체에서 발현하는 GTH subunit mRNA의 농도와 혈중 T및 혈중 $E_2$ 농도의 변화를 조사하였다. GnRHa를 체중 g당 $0.2\;{\mu}g$으로 복강 주사한 후 0, 6, 12, 24 및 48시간제에 뇌하수체와 혈액을 샘플링한 결과, 호르몬 처리 후 6시간째에 FSH$\beta$ mRNA가 유의적으로 증가하였으나, LH$\beta$ mRNA 발현에는 유의적인 차이가 없었다. 또한 혈중 T와 혈중 $E_2$ 농도는 GnRHa처리 후 24시간째 유의적으로 증가하였다.

• 한국수정란이식학회지
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• 제10권1호
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• pp.73-82
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• 1995

large scale production of cloned embryos requires the technology of multiple generation nuclear transplantation(NT) using NT embryos as the subsequent donor nuclei. The purposes of this study were producing the second generation cloned rabbit embryos, and also to determine the electrofusion rate and in vitro developmental potential comparatively in the cloned embryos of the first and second NT generation. The embryos of 16-cell stage were collected from the mated does by flushing oviducts with Dulbecco's phosphate buffered saline(D-PBS) containing 10% fetal calf serum(FCS) at 47 hours after hCG injection In the first generation NT, the nuclear donor embryos were synchronized in the phase of Gi /S transition of 32-cell stage. The first generation NT embryos which were developed to 8-cell were synchronized in Gi /S transition phase of the following 16-cell stage and used as donor nuclei for second generation Synchronization of the cell cycle of blastomeres was induced, first, using an inhibitor of microtuble polymerization, colcemid for 10 hours to arrest blastomeres in M phase, and secondly, using a DNA synthesis inhibitor, aphidicolin for 1.5 to 2 hours to arrest them in Gi /S transition boundary. The recipient cytoplasms were obtained by removing the nucleus and the first polar body from the oocytes collected at 14 hours after hCG injection. The separated donor blastomeres were injected into the enucleated recipient oocytes by micromanipulation and were electrofused by electrical stimulation of three pulses for 60 $\mu$sec at 1.25 kV /cm in 0.28 M rnannitol solution The fused oocytes were co-cultured with a monolayer of rabbit oviductal epithelial cells in M-199 solution containing 10% FCS for 120 hours at 39$^{\circ}C$ in a 5% $CO_2$ incubator. Following in vitro culture of the first and second generation cloned embryos to blastocyst stage, they were stained with Hoechst 33342 dye for counting the number of blastomeres by fluorescence microscopy. The results obtained were summarized as follows: 1. The electrofusion rate was found to be similar as 79.4 and 91.5% in the first and second generation NT rabbit embryos, respectively. 2. The in vitro developmental potential to blastocyst stage of the second generation NT embryos (23.3%) was found significantly(p<0.05) lower, compared with that of the first generation NT embryos (56.8%). 3. The mean blastomeres counts of embryos developed to blastosyst stage following in vitro culture for 120 hours and also their daily cell cycles during the culture period were decreased significantly (p<0.05) to 104.3 cells and 1.33 cylces in the second NT generation, compoared with 210.4 cells and 1.54 cycles in the first NT generation, respectively.

• 대한한방종양학회지
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• 제4권1호
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• pp.71-87
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• 1998

Even though appropriate immune response is necessary for the survival of the individual, excessive or insufficient immune Response might cause autoimmune or allergic disease. So the immune response must be controlled to the degree that is beneficial for the well being of the individual. This study was undertaken to know the effects of Gunleetang Gagambang on the immune system of the mouse. Gunleetang Gagambang 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 Gunleetang Gagambang 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(S180 cells). Treatment of the Gunleetang Gagambang on 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, Gunleetang Gagambang decreased not only the frequency of tumor production but also the number and the weight of tumors per tumor bearing mice(TBM). Gunleetang Gagambang on 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 Gunleetang Gagambang extract into TBM. In vitro, treatment of Gunleetang Gagambang 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. Gunleetang Gagambang also stimulated the migrative ability of leukocyte, the MIF and IL-2 production of T lymphocytes, but not IL 6 production of B cells. Gunleetang Gagambang administration to mice enhanced NK cells activities. These results demonstrated that Gunleetang Gagambang 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 Gunleetang Gagambang might be chiefly due to nonspecitie enhancement of NK cell activities and cell-mediated immune responses.

• Asian Pacific Journal of Cancer Prevention
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• 제16권18호
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• pp.8553-8557
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• 2016

Background: Accurate dose assessment and correct identification of irradiated from non-irradiated people are goals of biological dosimetry in radiation accidents. Objectives: Changes in the FDXR and the RAD51 gene expression (GE) levels were here analyzed in response to total body exposure (TBE) to a 6 MV x-ray beam in rats. We determined the accuracy for absolute quantification of GE to predict the dose at 24 hours. Materials and Methods: For this in vivo experimental study, using simple randomized sampling, peripheral blood samples were collected from a total of 20 Wistar rats at 24 hours following exposure of total body to 6 MV X-ray beam energy with doses (0.2, 0.5, 2 and 4 Gy) for TBE in Linac Varian 2100C/D (Varian, USA) in Golestan Hospital, in Ahvaz, Iran. Also, 9 rats was irradiated with a 6MV X-ray beam at doses of 1, 2, 3 Gy in 6MV energy as a validation group. A sham group was also included. After RNA extraction and DNA synthesis, GE changes were measured by the QRT-PCR technique and an absolute quantification strategy by taqman methodology in peripheral blood from rats. ROC analysis was used to distinguish irradiated from non-irradiated samples (qualitative dose assessment) at a dose of 2 Gy. Results: The best fits for mean of responses were polynomial equations with a R2 of 0.98 and 0.90 (for FDXR and RAD51 dose response curves, respectively). Dose response of the FDXR gene produced a better mean dose estimation of irradiated "validation" samples compared to the RAD51 gene at doses of 1, 2 and 3 Gy. FDXR gene expression separated the irradiated rats from controls with a sensitivity, specificity and accuracy of 87.5%, 83.5% and 81.3%, respectively, 24 hours after dose of 2 Gy. These values were significantly (p<0.05) higher than the 75%, 75% and 75%, respectively, obtained using gene expression of RAD51 analysis at a dose of 2 Gy. Conclusions: Collectively, these data suggest that absolute quantification by gel purified quantitative RT-PCR can be used to measure the mRNA copies for GE biodosimetry studies at comparable accuracy to similar methods. In the case of TBE with 6MV energy, FDXR gene expression analysis is more precise than that with RAD51 for quantitative and qualitative dose assessment.

• 한국식품영양과학회지
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• 제41권12호
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• pp.1701-1707
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• 2012

3,3'-Diindolylmethane(DIM)은 십자화과 채소에 포함되어 있는 indole-3-carbinol(I3C)이 동물의 산성 위액에서 중합되어 생성된 물질이다. 지금까지 DIM은 유방암, 전립선암 그리고 대장암 세포주에서 항암효과가 있다고 알려져 있으며 그 기작에 대한 연구도 다양하게 진행되고 있다. 그러나 정상세포에서 암화과정 중 암의 촉진과 진행과정의 주요한 항암 표적인 항염증에 대한 연구는 보고된 바 없다. 따라서 본 연구에서는 유방상피세포인 MCF-10A 세포에 12-O-tetradecanoylphorbol-13-acetate(TPA)로 염증반응을 유도한 후 DIM이 염증작용에 미치는 영향을 조사하였다. 그 결과 MCF-10A 세포에서 TPA에 의해 유도된 COX-2 단백질 및 mRNA 발현이 DIM 처리에 의해 현저히 감소하였다. 또한 TPA에 의해 유도된 $I{\kappa}B-{\alpha}$의 분해, p65의 핵으로의 이동, $NF-{\kappa}B$ DNA binding activity 역시 DIM의 처리에 의해 감소하였다. 이는 DIM이 인간의 유방상피세포인 MCF-10A 세포에서 TPA에 의해 유도된 $NF-{\kappa}B$ 신호전달체계의 활성을 억제함으로써 COX-2의 발현을 억제하여 염증반응을 조절함을 나타낸다. 그러므로 이러한 결과들은 DIM을 염증성 질환 예방제 또는 치료제로 개발할 수 있는 가능성을 시사한다.

• 한국임상약학회지
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• 제16권1호
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• pp.23-27
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• 2006

Doxorubicin (DXR) is a type of anti-cancer drug called an 'anthracycline glycoside', It works by impairing DNA synthesis, a crucial feature of cell division, and thus is able to target rapidly dividing cells. Doxorubicin is a very serious anti-cancer medication with definite potential to do great harm as well as great good. A liquid chromatography-tandem mass spectroscopy (LC-MS/MS) method was developed to identify and quantify DXR in small-volume biological samples. After the addition of internal standard (IS, $5{\mu}L\;of\;1{\mu}M/ml$ daunorubicin methanol solution) into the serum sample, the drug and IS were extracted by methanol. Following vortex for a 1min and centrifugation at 15,000g for 10 min the organic phase was transferred and evaporated under a vacuum. The residue was reconstituted with $350{\mu}L$ of mobile phase and $10{\mu}L$ was injected into C18 column with mobile phase composed of 0.05M ammonium acetate (0.1 M acetic acid adjusted to pH 3.5) and acetonitrile (40:60, v/v). The flow rate was kept constant at $350{\mu}L/min$. The ions were quantified in the multiple reaction mode (MRM), using positive ions, on a triple quadrupole mass spectrometer. The lower limits of quantification for Doxorubicin in plasma and small tissues were approximately 0.5 ng/mL and 0.5 ng/mL respectively. Intra- and inter-assay accuracy (% of nominal concentration) and precision (% CV) for all analytes were within 15%, respectively.

• Asian Pacific Journal of Cancer Prevention
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• 제17권sup3호
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• pp.155-160
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• 2016

Breast cancer (BC) is the second most common cancer in the world and by far the most frequent cancer among women, with an estimated 1.67 million new cancer cases diagnosed in 2012 (25% of all cancers). Polygene expression analysis is used to predict the prognosis and determine the most appropriate treatment regimen. The objective of this study was to examine the gene expression profiles of SIRT3, HRAS, LSP1, SCUBE2 and AP2A2 in Iranian women with BC.A total of 136 patients including healthy controls were categorized into three groups based on the relapse of the disease. Expression of desired genes in formalin-fixed, paraffin embedded tissues collected from all groups of participants was analyzed via the RT PCR method. RNA extraction and cDNA synthesis were performed then real-time quantitative PCR was carried out. Gene expression analysis revealed that the expression of SIRT3 was equal among patient and control groups. LSP1 was down regulated in all patient groups relative to controls but reduced expression in the metastatic group relative to the non-metastatic one was not significant. HRAS was significantly overexpressed in total and metastatic tumor samples versus normal but not in non-metastatic cases. SCUBE2 expression showed significant over-expression in both overall tumor samples and the non-metastatic group as compared to normal tissues. Gene expression level of AP2A2 in all groups was not detectable. Our data are compatible with a tumor suppressor role of LSP1 related to potential prognostic factor for tumor recurrence and outcome. This study for the first time assayed the prognostic value and changes in the expression of SIRT3, LSP1, HRAS, SCUBE2 and AP2A2 genes in women with breast cancer in the Iranian population and findings confirmed potential biomarker and prognostic capability of these genes. Such expression profiling data can critically improve prognosis and treatment decisions in cancer patients.

• 한방안이비인후피부과학회지
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• 제17권3호
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• pp.38-60
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• 2004

We made an experiment if the extracts of DanGuiBoHyulTangGami-Bang(DBTG) and 15 kinds of the medical herbs used the materials of DBTG were effective on the hair formation palpation and the falling out of hair, and came to the following conclusions. 1. The extracts of Paeonia lactiflora, Cuscuta chinensis and Angelica tenuissima of DBTG consisted of the 15 kinds of the medical herbs kept the activity of 5${\alpha}$-reductase type Ⅱ from being active 75.3$\%$, 63.8$\%$. 75.5$\%$. 2. The hair growth index, 1.6(control group 0.8) of the extracts of DBTG bas a little effect on the hair growth palpation and that of Rubus coreanus 1.8(control group 0.4) was the most effective one of the medical herbs, and Paeonia lactiflora 2.3(control group 1.7) and Vitex rotundifolia 2.3(control group 1.5) showed the effect on hair formation palpation. 3. The hair growth period couldn't be extended by DBTG in this experimental stage. 4. The 15 kinds of constitution medicines of DBTG didn't have effects in dermal papilla cells DNA increase, IGF- I, KGF, HGF the revelation of a gene heredity, the protein synthesis of the hair follicle tissues. 5. All of the 15 kinds of constitution medicines of DBTG didn't have the antibacterial activity in Paper disc rule. 6. The results from the test of a radical scavenging activity of the 15 kinds of constitution medicines of DBTG showed that the extracts of Paeoria lactiflora, Scutellaria baicalensis, Rubus coreanus have the superior antioxidant activity in the concentration of 0.01$\%$ and 0.001$\%$ 7. In the formation controlled experiment, Vitex rotundifolia (70.6$\%$), Scutellaria baicalensis (47.1$\%$, Saposhnikovia (44.8$\%$) of the 15 kinds of constitution medicines of DBTG in the 50㎍／㎖ concentration controled NO forming and Vitex rotundifolia (12.7$\%$) controled NO forming in the 5㎍／㎖ concentration in order. 8. MTT(lC／50) of the extracts of Rehmannia glutinosa, Paeonia lactiflora, Scutellaria baicalensis, Lycium chinense, Rubus coreanus of the 15 kinds of constitution medicines of DBIG was more than 500㎍／㎖ and had the least cell virulence.