• 동의생리병리학회지
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• 제28권3호
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• pp.310-316
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• 2014

Oldenlandia diffusa, Cremastra appendiculata and Fritillaria thunbergii are widely distributed in the Korea, China and Japan, and has been used in traditional medicine for various diseases, such as pharyngolaryngitis, tonsillitis, goiter and stomach ulcer. However, the anti-cancer activities in human breast cancer have not been clearly elucidated yet. In this study, it was compared the in vitro cytotoxic effects of single and complex treatment of O. diffusa, C. appendiculata and F. thunbergii. We treat human breast cancer MCF-7 cells with O. diffusa, C. appendiculata and F. thunbergii. And we evaluated viability, growth inhibition, morphological changes, apoptotic bodies formation, measurement of the cell cycle and formation of DNA fragmentation of these cells. It was found that single treatment of O. diffusa could inhibit the cell proliferation in human breast cancer MCF-7 cells. However, complex treatment of O. diffusa, C. appendiculata and F. thunbergii is weakly or not affect the cell proliferation of MCF-7 cells. And anti-proliferative effects of O. diffusa in MCF-7 cells was associated with G1 arrest of cell cycle. These findings suggest that O. diffusa may be a potential chemotherapeutic agent for the control of human breast cancer cells and further studies will be needed to identify the molecular mechanisms.

• 대한한방내과학회지
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• 제35권2호
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• pp.133-144
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• 2014

O. diffusa, C. appendiculata and F. thunbergii are reported to possess many pharmacological activities including anti-oxidant, anti-inflammatory, anti-hypertension, anti-diabetic and anti-cancer effects. However, their anti-cancer activities in human breast cancer have not been clearly elucidated yet. Objectives: In the present study, we compared the in vitro cytotoxic effects of single and complex treatment of O. diffusa, C. appendiculata and F. thunbergii in human breast cancer MDA-MB-231 cells. Methods: After we treated human breast cancer MDA-MB-231 cells with O. diffusa, C. appendiculata and F. thunbergii. we evaluated viability, growth inhibition, morphological changes, apoptotic body formation, measurement of the cell cycle and formation of DNA fragmentation of these cells. Results: We found that single treatment of O. diffusa and F. thunbergii could inhibit cell proliferation in human breast cancer MDA-MB-231 cells. However, complex treatment of O. diffusa, C. appendiculata and F. thunbergii had weak or no effect on the cell proliferation of MDA-MB-231 cells. The first, anti-proliferative effects of O. diffusa in MDA-MB-231 cells was associated with G2/M arrest of cell cycle and apoptotic cell death. The second, anti-proliferative effect of F. thunbergii in MDA-MB-231 cells was associated with apoptotic cell death. Conclusions: Taken together, these findings suggest that O. diffusa and F. thunbergii may be a potential chemotherapeutic agent for the control of human breast cancer cells, further studies will be needed to identify the molecular mechanisms.

• 식물분류학회지
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• 제41권1호
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• pp.58-65
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• 2011

우리나라 미기록 종인 닭의장풀과(Commelinaceae)의 고깔닭의장풀(Commelina benghalensis L.)과 큰닭의장풀(Commelina diffusa Burm. f.) 2종이 제주도의 저지대에서 채집되었다. 고깔닭의장풀은 불염포의 밑 부분이 합생하여 깔때기 모양을 하고 폐쇄화를 갖는다는 점에서 나머지의 다른 종들과 구분이 된다. 큰닭의장풀은 삭과가 3개의 과피편으로 되어 있고, 불염포상 총포편이 피침형이면서 심장저 또는 원저라는 점에서 나머지 다른 종들과 구분된다. 염색체수는 고깔닭의장풀이 2n = 2x = 22(2배체)이며, 염색체 크기가 1.25-2.70 ${\mu}m$로 매우 작았고, 큰닭의장풀은 2n = ca. 100 이상으로 정확한 수를 알 수 없었다. 이 종들은 지금까지는 아시아와 아프리카의 열대 및 아열대에 분포하여 우리나라보다 남쪽에만 분포하는 것으로 알려져 왔다.

• 대한한방종양학회지
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• 제6권1호
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• pp.81-97
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• 2000

Objectives: This experimental study was carried out to evaluate the effects of aqueous and methanol extracts of Hedyotis diffusa which has long been used for cancer treatment in oriental medicines on the induction of apoptotic cell death in human lymphoid leukemia cell line, HL-60. Methods: Cells were treated with various concentrations (200 to $0.4{\mu}g$) and periods (6 to 30 hr) of $H_2O$ and methanol extracts of Hedyotis diffusa. Then, cells were tested for viability by MTT assay. Cells wrere treated with $200{\mu}g/ml$ of methanol extract fork various periods. Genomic DNA was isolated, separated, on 1.5% agarose gels, stained with ethidium bromide and visualized under UV light. Cells were treated with $200{\mu}g/ml$ of each extract for 16 hr. Then, cells were treated with Hoechst dye 33342 and observed by fluorescence microscopy. Cells were treated with various doses of each for 12 hr and $100{\mu}g/ml$ of methanol extract for various periods. Lysate from the cells used to measure the activity of Caspase-1 and-3 proteases by using fluorogenic peptide substrates including acetyl-YVAD-AMC and acetyl-DEVD-AMC, respectively. Cells were treated with $200{\mu}g/ml$ of each extract for various periods. Cell lysates were immunoprecipated with anti-JNKl antibodies. The immune complex was reacted with $32^p-ATP$ and c-Jun as a substrate. The phosphotransferase activity of JNKI was measured by using PhosphoImage analyzer (Fuji Co., Japan). Nuclear extracts were isolated and incubated with oligonucleotide probe of $NF-{\kappa}B$. Transcriptional activation of ${\kappa}B$ was measured by using EMSA and visualized by PhosphoImage analyzer (Fuji Co, Japan). Cell lysates were prepared and analyzed by Western blotting with anti-Bc12 antibodies and anti-Bax antibodies. Cells were pretreated with various doses of methanol extract for 2 hr. Then, the extract was removed by centrifugation. Cells were resuspended with RPMI-1640 media containing 0.3% agarose, 10% FBS, overlayred onto bottom layer agarose and incubated at $CO_2$ incubator for 6 days. The number of colony was counted under light microscopy ($\time100$). Results: The death of HL-60 cells was markedly induced by the addition of methanol extract of Hedyotis diffusa in a dose and time-dependent manners. The apoptotic characteristic ladder pattern of DNA strand break was observed in death of HL-60 cells. In addition, it was shown nucleus chromatin condensation and fragmentation under Hoechst staining. Therefore, Hedyotis diffusa extract-induced death of HL-60 cells is mediated by apoptotic signaling processes. The activity of Caspase 3-like proteases remained in a basal level in HL-60 cells treated with aqueous extract of Hedyotis diffusa. However, it was markedly increased in HL-60 cells treated with methanol extract of Hedyotis diffusa. In addition, the phosphotransferase activity of JNKl was increased in HL-60 cells treated with methanol extract of Hedyotis diffusa. Furthermore, the activation of transcriptional activator, $NF-{\kappa}B$ was markedly induced by methanol extract of Hedyotis diffusa. Anti-apoptotic Bc12 was cleaved into 23Kda fragment by treatment of methanol extract of Hedyotis diffusa. However, expression of proapoptotic Bax protein was increased by treatment of methanol extract of Hedyotis diffusa in a time-dependent manner. Furthermore, methanol extract markedly inhibited the colony forming efficiency of HL-60 cells in semisolid agar culture. Conclusions: Above results suggest that methanol extract of Hedyotis diffusa induces the apoptotic death of human leukemic HL-60 cells via activations of Caspase-3 proteases, JNKI, transcriptional activator $NF-{\kappa}B$, In addition, our results also suggest that methanol extract of Hedyotis diffusa reduces the malignant potential of HL-60 cells via down regulation of colony forming effciency through cleavage of Bc12 as well as induction of Bax.

• 대한한방종양학회지
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• 제6권1호
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• pp.67-79
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• 2000

Objective : Hedyotis diffusa has been used as an anticancer agent for several decades in oriental medicine. We test whether the methanol extract of the herb affects transcriptional activation factors including $NF-{\kappa}B$ and AP-1. Methods : 1. HL-60 cells were treated with various concentrations(from 200 to $50{\mu}g/ml$) of methanol extract and $H_2O$ extract($200{\mu}g/ml$)of hedyotis diffusa, After 48h later, the cells were tested for viability by MTT assay. 2. The HL-60 cells were treated with $200{\mu}g/ml$ of methanol extract for the indicated periods. First. Nuclear extracts were isolated and incubated with oligonucleotide probe of $NF-{\kappa}B$ and AP-1. Second. Nuclear extracts were isolated and reacted with p50, p65. c-rel pan-Jun, c-Jun, JunB. JunD antibody on ice for 30min. Finally The cell lysates were prepared and analyzed by western blotting using anti-Fas, anti-FasL and anti-p53 antibody. Results : 1. The methanol extract decreases the viability of human lymphoid origin leukemia HL-60 cells in a dose-dependent manner. 2. $NF-{\kappa}B$ is rapidly activated by the addition of the methanol extract, reaches a peak at 30min and gradually returns to resting level. We confirm that $NF-{\kappa}B$ is a heterodimer mainly composed of p65 subunit with c-Rel. 3. Transcriptional activation of AP-1 is detected at 30min and reaches a maximum at 1hr after stimulation of the cells with the methanol extract. AP-1 is mainly composed with Jur-D and partially Jug-B proteins. 4. the methanol extract of Hedyotis diffusa induces the expression of Fas, Fas ligand and p53 proteins of HL-60 cells in a time dependent fashion. Conclusions : These results suggest that the methanol extract of Hedyotis diffusa exerts anticancer effects to induce the death of human leukomic HL-60 cells via activation of trascriptional factors such as $NF-{\kappa}B$ and AP-1, increase in expression of Fas mediated signalling proteins, and induction of tumor suppressor gene. p53.

• 식물분류학회지
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• 제51권1호
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• pp.10-17
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• 2021

Invasive alien species management is pivotal for biodiversity conservation. Commelina caroliniana Walter, from the family Commelinaceae, is an alien plant native to the Himalayas and India, but it has been widely introduced around the world, including in the United States, Brazil, Philippines, and Japan. In Korea, the first population was found growing adjacent to agricultural land and farm roads on Jejudo Island, and field observations confirmed the presence of at least nine populations there. It is similar morphologically to C. diffusa Burm. f. but can be distinguished by involucral bracts that are ciliate at the base, hairs on the peduncle and obsolete upper cincinnus, brown spots on its 4-lobed antherode, and seed surfaces that are smooth to slightly alveolate. It was determined to have an invasiveness low score of 8 according to the Korean 'Invasive Alien Plant Risk Assessment', suggesting that it may spread to natural habitats. Although the current distribution of C. caroliniana is restricted to Jeju-si, it has spread dramatically in many other areas of the world. At present, it has had a limited impact on the local environment, but local and regulatory authorities should pay close attention to this plant and take measures to prevent its expansion in the future.

• 한국토양비료학회지
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• 제45권4호
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• pp.602-609
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• 2012

Burkholderia anthina R-4183, Burkholderia diffusa R-15930 and Burkholderia stabilis LMG 14294 isolated from green house soils (Gongju-Gun area, South Korea) were characterized and their phosphate solubilizing ability was assessed. Under in vitro culture conditions, all three species were proved to be effective in solubilizing phosphates in varying degrees. Strain Burkholderia anthina exhibited the highest phosphate solubilization in NBRIP medium ($665{\mu}g\;ml^{-1}$) followed by Burkholderia diffusa ($630{\mu}g\;ml^{-1}$) and Burkholderia stabilis ($578{\mu}g\;ml^{-1}$). However, solubilization of $FePO_4$ and $AlPO_4$ was found to be poor in all the strains. Acidification by means of gluconic and oxalic acids accumulation in the culture medium could be the possible mechanism responsible for phosphate solubilization. Glucose at the rate of 3% was found be the best carbon source for Burkholderia anthina while other two Burkholderia species showed maximum phosphate solubilization at 2% of glucose. In the case of nitrogen sources, ammonium and nitrate were equally effective in solubilizing phosphates by Burkholderia species. Despite a slight decrease in phosphate solubilization observed at increasing temperature, all three Burkholderia species could withstand a temperature of $30-35^{\circ}C$, pH at the range of 7-9 and the presence of NaCl (up to 2.5%) without much compromising the phosphate solubilization. As shown with potted mung bean seedlings, all the three isolates could enhance soil fertility and plant growth indicating their great potential to be used as bio-inoculants.

• International Journal of Industrial Entomology and Biomaterials
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• 제18권1호
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• pp.49-52
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• 2009

An in vitro and in vivo studies were conducted to evaluate the antibacterial efficacy of certain botanicals viz., rhizomes of turmeric (Curcuma longa) and leaves of amla (Phyllanthus emblica), asparagus (Asparagus racemosus), bael (Aegle marmelos), boerhavia (Boerhavia diffusa), garlic (Allium sativum) and basil (Oscimum basicilum) against bacterial pathogens viz., Staphylococcus sp., Bacillus sp. and Klebsiella cloacae, of silkworm, Bombyx mori. Asparagus and basil, amla and boerhavia, basil and bael at concentration of 20, 000 ppm showed higher antibacterial activity against Staphylococcus sp., Bacillus sp., K. cloacae respectively, both in vitro and in vivo studies.

• 생명과학회지
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• 제23권8호
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• pp.1025-1031
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• 2013

백운풀의 품질표준화를 위하여 지표 성분을 설정하고자 하였다. 백운풀의 에탄올 추출물 중 비교적 백운풀에 특이하고, 다량 함유되어 있는 화합물을 분리한 다음 $^1H$ 및 $^{13}C$-NMR을 이용하여 구조 동정한 결과, HD1 (digitolutein), HD2 (2-hydroxy-3-methylanthraquinone), HD3 [(E/Z)-6-O-p-coumaroyl scandoside methyl ester, 4:1] 및 HD4 [(E/Z)-6-O-p-methoxycinnamoyl scandoside methyl ester, 4:1]의 4 종 화합물을 표준품으로 확보할 수 있었다. HD1~4의 검량선을 작성한 결과 상관계수($R^2$)는 각각 0.9991, 0.9999, 0.9993, 0.9998로 높은 직선성을 보였으며, LOD는 0.05, 0.06, 0.03, $0.07{\mu}g/ml$로, LOQ는 0.165, 0.198, 0.99, $0.231{\mu}g/ml$ 수준으로 나타났다. 또한 면적 비에 대한 재현성 RSD (%)는 각각 0.16, 0.90, 1.18, 0.58%의 값을 보였으며 일내 CV (%)는 0.23, 2.00, 1.18, 0.26%를, 일내 정확도는 102.80, 96.08, 108.44, 94.60%로 나타났다. 한편, 일간 CV (%)는 각각 0.25, 1.16, 0.98, 0.30%, 일간 정확도는 102.82, 98.58, 110.23, 94.73%였다. 백운풀 추출물 중 설정한 4 종 지표성분의 정량을 위한 최적 HPLC 조건을 검토한 결과, 고정상은 Merck Chromolith RP-18e ($100{\times}4.6mm$, $5{\mu}m$), 컬럼 온도는 실온, 검출은 UV 280 nm, 유속 2.0 ml/min, injection volume $10{\mu}l$, 이동상으로는 0.5% acetic acid가 포함된 $H_2O$ (A)와 methanol (B)을 사용하여 최초 80% (A)로 시작하여 9분 후 40% (A)가 되도록 gradient를 준 다음 18분까지 유지하는 조건을 설정하였다. 이 방법으로 백운풀 ethanol 추출물을 분석할 경우 다른 성분의 방해 없이 20분 이내에 모든 지표성분을 분석할 수 있었다. 연구결과는 백운풀의 건강기능식품 및 천연물신약 개발 및 품질관리에 도움을 줄 수 있을 것으로 기대된다.

• 동의생리병리학회지
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• 제18권2호
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• pp.408-412
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• 2004

For the evaluation of immunomodulatory effect of Korean Oldenlandiae Herb (OH) and Radix (OR), our experiment was performed with methanol extracts of Korean Oldenlandiae Herba and Radix. After administration of methanol extracts of Korean OH and OR for 7 days, Balb/C mice were immunized with sheep red blood cells. Four days later, splenic leukocytes were isolated and immunological experiments were performed. Rosette forming cells and plaque forming cells were significantly increased in Korean OH and OR treated mice compared with PBS treated control. Korean OH and OR also enhanced T and B lymphocytes, macrophage and natural killer cells by flowcytometric analysis. LPS-induced TNF-α and IL-6 levels were increased by OH and OR compared with untreated control. These results suggest that Korean OH and OR have immunomodulatory activity through regulation of cell-mediated immune and humoral immune response.