• Journal of Sericultural and Entomological Science
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• v.25 no.1
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• pp.1-20
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• 1983

It has been known that the insect molting hormone and its analogues exist also in plant kingdom and their concentration has been found to be about 0.1~2.0% of dry matter, which is equivalent to $10^3{\sim}10^5$ times of those in insects. This study was carried out; 1) to isolate the phytoecdysones from Korean Achyranthes radix and characterize their physico-chemical properties. 2) to investigate the biological activity of this phytoecdysone on Bombyx mori larvae. The resuls were summarized as follows; 1. The extraction method of phytoecdysones was optimized by three consecutive reflux for 1hr using 200g of dried and milled radix per 1l methanol. 2. The purification from the crude extract was made by a series of steps such as precipitation of gum-type polymer with n-Butyl acetate, adsorption on technical grade silica and chromatography with neutral alumina. The conditions of each step were optimized and the resulting crude crystal was about 500mg per kg dry radix. 3. The crude crystal from the cultivated Achyranthes(Achyranthes japonia) contained ecdysterone (20-hydroxyecdysone) and inokosterone in the proportion of one to one. In order to separate these, a series of processes such as acetylation, separation by alumina column chromatography deacetylation by alcoholysis, deionization and crystallization were introduced and optimized 125mg of ecdysterone and 18mg of inokosterone per kg dry radix were thus obtained. 4. The wild Achyranthes (Achyranthes obtusifolia) radix was found to contain the ecdysterone only. A 285mg of ecdysterone was crystallized per kg dry radix. 5. Isolated ecdysterone, inodosterone and acetylated compounds were characterized by IR., UV., NMR spectroscopy, mp, TLC and densitometry. 6. Ligation experiment was undertaken to confirm the biological activity of the purified ecdysterone; the ecdysterone could induce larval-pupal metamorphosis in the ligated abdomen of 4th instar larvae injecting 0.5~1.0${\mu}g$. 7. By ecdysterone feeding experiment using artificial diet, it was elucidated that the critical time of feeding would be the first half of each instar resulting in increased weight of silk layer. 8. The ecdysterone was fed to 5th instar silkworm at the level of 1, 2, 3, 5ppm of dry feed of artificial diet containing 5% mulberry leaves for 72hrs. At 2ppm of the concentration. body weight and silk layer weight were arrived at maximum. But at higher concentrations body weight and silk layer weight decreased than the control group. At 2ppm of the concentration, body weight was increased by 12.5%. 9. Feeding 2ppm of ecdysterone at the later half of 5th instar, the duration of larvae was shortened.

• YAKHAK HOEJI
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• v.45 no.5
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• pp.455-459
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• 2001

Melandrii Herba has been used for tympanitis, sore throat, dysentery and dehumidifying diuretic. From the BuOH fraction of methanol extracts, three ecdysteroids (ecdysterone, ecdysterone 22-acetate and inokosterone) were isolated by column chromatography using Amberlite XAD-4, ODS and Sephadex LH-20 gel and by HPLC method. The structures of these compounds were identified on the basis of spectroscopic methods.

• Journal of Sericultural and Entomological Science
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• v.13 no.2
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• pp.135-139
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• 1971

It is very important to make mounting numerous silkworms by their normal growth when considering the sericultural management. There were several studies on the utilization of repellent to the matured silkworms to mounting, studies concerning forcing maturity of silkworms were developed considerably after studies on the insect ecdysis hormone. Commercial named MSH(II) (Inoksterone) similar substance of ecdysterone and DAT repellent (Dodesil alcohl) were used for this study, and the results can be summarized as follows; 1. MSH (II) mixed with the mulberry leaves shown the matured effect on the growth of silkworms and increased amount of MSH (II) shortened the last stage in their growth. 2. MSH (II) mixed with the mulberry leaves when 5% of early matured silkworms appeared were most effective. 3. DAT was hastened their mounting. 4. Utilization of MSH (II) and DAT together were hastened silkworms mounting and those chemicals can be used practically in the silkworm rearing.

• Korean Journal of Environmental Biology
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• v.23 no.3 s.59
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• pp.275-280
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• 2005

Development of the fourth-instar larvae of Chironomus riparius has a sensitive to ecdysteroid hormones. The 2D/E gel analysis for polypeptide expression reflecting early-ecdysterone inducible gene has conducted the emerged female from larval phase exposure to bisphenol A (BPA). In the 2D/E gel 1108 protein spots were identified. The visualized protein spots allowed extraction of 17 protein spots differed more than 3 fold in BPA treated animals, which was approximately $1.6\%$ of the total protein spots. However, polypeptide expression reflecting early-ecdysterone inducible gene didn't change after treatments. In addition, detection for the damages or changes in mitogenome level was observed. The conserved cytochrome oxidase I in DNA level affected exposure to BPA $(1{\mu}gL^{-1})$ in this preliminary study.

• Analytical Science and Technology
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• v.25 no.4
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• pp.250-256
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• 2012

Two marker compounds of Achyranthis Radix, ecdysterone and inokosterone, were analyzed by HPLC on an ODS column ($250{\times}4.6$ mm, 5 ${\mu}m$) with a mobile phase of 15% acetonitrile containing 0.08% formic acid at a flow rate of 1.0 mL/min and a detection wavelength of UV 254 nm. The method was validated by ICH guideline and applied to the monitoring of marker compounds in 93 samples of Achyranthis Radix collected at various areas in Korea and China. The new content criteria of ecdysterone and inokosterone, established using linear regression method were 0.033% and 0.020%, respectively. When the new content criteria were applied to the quality control test of commercial Achyranthis Radix, 95.4% of total samples including 100% of Korean and 92.6% of Chinese samples were passed the test. Application of new content criteria could protect the Korean products and decrease the distribution of Chinese products with lower quality.

• Korean Journal of Environmental Biology
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• v.23 no.3 s.59
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• pp.269-274
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• 2005

The 2D/E gel analysis for polypeptide expression reflecting I-18 C gene (early-ecdysterone inducible gene) has conducted the emerged C. riparius adults from larval phase exposure to tebufenozide acting as an ecdysteroidal molting hormone. Control group, the amount of ORE II of the I-18 C gene was larger than that of ORE I of this gene. After treatments, ORE I of the I-18 C gene was overexpressed as the polypeptide, whereas ORF II of this gene was expressed as the polypeptide and was clearly reduced expression. Accordingly, we consider that tebufenozide exhibited endocrine disruptions related processing of ecdysteroid receptor protein reflecting ORF II of I-18 C gene. Also, earlier emergence day was related overexpressed polypeptide reflecting ORE I of I-18 C gene. In this study result, tebufenozide induced changing of physiological condition, and then polypeptide expression reflecting early-ecdysterone inducible I-18 C gene was different between control group and exposure group.

• Natural Product Sciences
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• v.24 no.4
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• pp.241-246
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• 2018

The herbs of Lamium takesimense Nakai (Lamiaceae) is used to treat spasmodic and inflammatory disease. The four polar compounds, ecdysterone, isoacteoside, rutin and lamiuside C, were isolated and identified from the BuOH fraction of the L. takesimense MeOH extract. HPLC quantification was performed on a Capcell Pak C18 column ($5{\mu}m$, $4.6mm{\times}250mm$) with a gradient elution of $H_2O$ and 0.05% acetic acid in MeOH. The HPLC method was validated in terms of linearity, sensitivity, stability, precision, and accuracy. The quantitative level in plant material was determined as the following order: lamiuside C (4, 3.75 mg/g dry weight) > ecdysterone (1, 1.93 mg/g) > isoacteoside (2, 1.32 mg/g) > rutin (3, 0.97 mg/g).

• International Journal of Industrial Entomology and Biomaterials
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• v.10 no.2
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• pp.137-141
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• 2005

Though many insecticides are commercially available, development of resistance, pest resurgence and effects on non-target organisms led to the search for alternate insect pest management (IPM) strategy based on larval growth and reproductive fitness. Reproductive potential of insects depends on its acquiring of vitellogenic competence which is under hormonal control. Exogenous application of analogues of JR (JHAs) and ecdysterone could derail normal development and reproduction in insects by manipulating an array of physiological processes. In the rice pest, brown planthopper, Nilaparvata lugens, JHA, hydroprene induced metathetely from the fifth (final) instar nymphs in an age-dependent manner. Day 0 of the final instar showed highest sensitivity to induce this abnormal development. Adults emerged from treated day 3 nymphs looked normal. Both the morphotypes were reproductively incompetent and showed partial to complete sterility. Pre-adult exposure of the ovarian tissue to hydroprene suppressed mitotic division of germinal cells and induced abnormalities in the later s1ages of growth and differentiation of ovary in N. lugens. More over the nymphal exposure to hydroprene inhibited patency changes of follicular epithelium and affected competence of the follicles for yolk sequestration. In the absence of ovarian growth and oocyte differentiation, germarium found disintegrated, trophic core regressed and terminal oocytes resorbed. Hydroprene exposure to newly ecdysed brachypterous females did not affect ovarian development and egg production. Proper larval-adult transition appeared as a. prerequisite for vitellogenic competence in N. lugens for which the ovarian tissues must be exposed to ecdysterone in the internal milieu devoid of JH.

• Journal of Applied Biological Chemistry
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• v.58 no.1
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• pp.13-19
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• 2015

The roots of Achyranthes japonica Nakai were extracted with 100% aqueous and concentrated subfraction was separated with ultra-performance liquid chromatography-based activity profiling. Three compounds were isolated from the subfraction 5 through the repeated prep- high performance liquid chromatography column chromatography. According to the results of physico-chemical and spectroscopic data including NMR and MS, the chemical structures of the compounds were determined as ecdysterone (1), 25S-inokosterone (2), and 25R-inokosterone (3). Three phytoecdysones were showed weak inhibitory activity for thymus and activation-regulated chemokine expression levels in tumor necrosis factor (TNF)-${\alpha}$ plus IFN-${\gamma}$ induced HaCaT cells, respectively. However, those compounds 1-3 were exhibited the most potent inhibition (80-95% at $200{\mu}g/mL$) against TNF-${\alpha}$ expression levels in A23187 plus phorbol-myrisrate acetate-induced RBL-2H3 cells. As result, 100% aqueous extract of A. japonica has an excellent anti-atopy activity. It could be used to a large range of functional anti-atopy cosmetics.

• YAKHAK HOEJI
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• v.41 no.6
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• pp.709-713
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• 1997

For the investigation of bioactive natural products with hypoglycemic effect, we have evaluated various extracts of Pulsatillae Radix(Ranunculaceae), which has been used in traditional medicine for hematochezia due to intense evil heat, malaria, chills and fever, epistaxis and internal hemorrhoids. The ethylacetate extract of the radix of this plant was found to show a significant hypoglycemic effect on alloxan diabetogenic mice. Using bioactivity-guided chromatographic purification of the ethylacetate extract, hypoglycemic constituent: 2${\beta}$, 3${\beta}$, 14${\alpha}$, 20, 22R, 25-hexahydroxy-cholest-7-en-6-one was isolated and structurally identified by physico-chemical properties and spectroscopic evidences.