Masayuki Mikage

Kanazawa University, Kanazawa, Ishikawa, Japan

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Publications (48)38.01 Total impact

  • Taketo Okada, Masayuki Mikage, Setsuko Sekita
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    ABSTRACT: The cDNAs (Espals) encoding phenylalanine ammonia-lyase (PAL) were cloned from Ephedra sinica by reverse transcription polymerase chain reaction (RT-PCR) using degenerate primers and by 5' and 3'-rapid amplification of cDNA ends (RACE). 2166 bp of the open reading frame (ORF) encoded 722 amino acids; sequence analyses of Espal clones suggested that at least four isoforms of EsPAL (EsPAL1, 2, 3, 4) existed, with nine amino acids substitution in their sequences. Phylogenetic analysis of EsPAL and PALs from other plant species revealed that EsPAL and Pinus PAL formed a gymnosperm-type PAL subfamily. The recombinant EsPAL1 to 4 functionally catalyzed a PAL reaction and their K(m), V(max), K(cat) and K(cat)/K(m) values did not show significant differences. Semi-quantitative RT-PCR analysis indicated that the expression of Espal genes in the roots was higher than in the plant's aerial parts. In addition, the activity of PAL in the roots was also higher than in the aerial parts. These results suggest that Espal genes are expressed in the whole plant but are dominant in the roots rather than in the aerial parts.
    Biological & Pharmaceutical Bulletin 01/2009; 31(12):2194-9. · 1.78 Impact Factor
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    ABSTRACT: TLC and HPLC were used to identify possible chemical markers for evaluating the quality of the crude drug "Pogostemoni herba" (aerial part of Pogostemon cablin), which is a component of Kampo medicines. In addition to the reported patchouli alcohol and 2-hydroxy-6-methyl-3-(4-methylpentanoyl)-4-pyrone, three phenylethanoids were isolated from this plant material for the first time: acteoside, isoacteoside, and crenatoside. The usefulness of these compounds as indicators of the crude commercial drug under various TLC conditions was examined, and patchouli alcohol was found to give a definite spot with a reproducible Rf value. Therefore, we propose TLC of the methanol (MeOH) extract using patchouli alcohol as a marker as a convenient method for identifying the crude drug Pogostemoni herba.
    Yakugaku zasshi journal of the Pharmaceutical Society of Japan 01/2009; 128(12):1833-7. · 0.31 Impact Factor
  • Kampo Medicine 01/2008; 59(2):279-285.
  • Masayuki MIKAGE, Hiroko ENDO
    Kampo Medicine 01/2008; 59(1):25-34.
  • Taketo Okada, Masayuki Mikage, Setsuko Sekita
    Biological & Pharmaceutical Bulletin 01/2008; 31(12):2194-2199. · 1.78 Impact Factor
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    ABSTRACT: Plants of Ephedra normally have vessels, but are known to become nearly vessel-less in some alpine localities. Previous studies implied that wood formation in Ephedra differs fundamentally from that in dicotyledons in which vessel-bearing and vessel-less taxa are systematically distinct. Using E. pachyclada in the Mustang district of Nepal, growing in an altitudinal range of over 2000 m, variation in wood formation and adaptation to alpine environment was studied in this normally vessel-bearing species. Variation in wood anatomy and wood formation was observed with conventional optical microscopy. The lengths of three kinds of tracheary elements were measured and statistically analysed against habitat altitude and plant size of the individuals studied. In E. pachyclada three kinds of tracheary elements, vessel elements, tracheids and fibre-tracheids, were nearly equal in length within individuals showing no elongation after differentiation from cambial initials. Tracheary element lengths among individuals had a negative correlation with altitude and a positive correlation with plant size. Multivariate analyses showed that altitude has a stronger correlation with tracheary element lengths than plant height or stem diameter. Moreover, several individuals from high elevations completely lacked vessels, and vessel formation fluctuated even in individuals from lower elevations. Wood anatomical trends in E. pachyclada are considered as an adaptation to extremely dry conditions in high mountains. Fluctuation in vessel formation in individuals from low elevations indicated that vessels differentiate only when their lateral expansion is allowed. These results showed that E. pachyclada has a different system of wood formation from dicotyledons and supported the opinion that the wood structure of Gnetales is fundamentally different from that of angiosperms.
    Annals of Botany 09/2007; 100(2):315-24. · 3.30 Impact Factor
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    Toshiyuki Atsumi, Nobuko Kakiuchi, Masayuki Mikage
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    ABSTRACT: We determined the DNA sequences of the internal transcribed spacer 1 and 2 (ITS 1 and 2), the 5.8S rRNA gene and most of the 28S rRNA gene of Poria cocos for the first time, and conducted analysis of 20 samples including cultured mycelias and crude drug materials obtained from various localities and markets. Direct sequencing of the ITS 1 and 2 regions of the samples, except for four wild samples, showed that they had identical DNA sequences for ITS 1 and 2 with nucleotide lengths of 997 bps and 460 bps, respectively. By cloning, the four wild samples were found to have combined sequences of common ITS sequences with 1 or 2-base-pair insertions. Altogether both ITS 1 and 2 sequences were substantially longer than those of other fungal crude drugs such as Ganoderma lucidum and Polyporus umbellatus. Thus, Poria cocos could be distinguished from these crude drugs and fakes by comparing the nucleotide length of PCR products of ITS 1 and 2. Contrary to the basic homogeneity in ITS 1 and 2, three types (Group 1, 2, 3) of the 28S rRNA gene with distinctive differences in length and sequence were found. Furthermore, Group 1 could be divided into three subgroups depending on differences at nucleotide position 690. Products with different types of 28S rRNA gene were found in crude drugs from Yunnan and Anhui Provinces as well as the Korean Peninsula, suggesting that the locality of the crude drugs does not guarantee genetic uniformity. The result of DNA typing of Poria cocos may help discrimination of the quality of the crude drug by genotype.
    Biological & Pharmaceutical Bulletin 09/2007; 30(8):1472-6. · 1.78 Impact Factor
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    ABSTRACT: More than 400 species of medicinal plants grow in the Northern Areas of Pakistan, including Ephedra plants. To investigate the wild Ephedra plant resources in the area, we surveyed the medicinal plants and collected 71 specimens from 18 collecting sites to analyze their genetic variation. The DNA sequences of the internal transcribed spacers 1 and 2 (ITS 1 and 2) of nuclear ribosomal DNA and a noncoding sequence of chloroplast DNA (trn L/F) were analyzed. This DNA data analysis and external morphological features were used to confirm the species of the specimens, and it was found that E. intermedia was the major species in the area and that E. gerardiana and E. przewalskii were present sporadically. Although it inhabits a relatively small area in comparison with the northwestern Chinese provinces, the DNA sequence of E. intermedia in the Northern Areas of Pakistan was significantly more heterogeneous than the same species grown in those neighboring regions. Most of the E. intermedia specimens contained more than 0.7% ephedrine alkaloids, fulfilling the requirement of the Japanese Pharmacopoeia; thus, the Ephedra plants in the area are a genetic and medicinal resource of great importance.
    Journal of Natural Medicines 06/2007; 61(3):357-365. · 1.45 Impact Factor
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    ABSTRACT: The tuberous root of aconite, Aconiti tuber, contains tonic but poisonous aconite alkaloids, such as aconitine, hipaconitine and mesaconitine. The standardization of the cultivation protocols must be established, since the quality of the crude drug, especially the content of aconite alkaloids, is influenced by cultivation conditions. Here, we report the effects of planting depth, disbudding and root pruning on the content of the aconite alkaloids as well as on the weight of the root tuber of aconite. The daughter root tubers of shallow-planted plants showed a significantly higher content of aconitine, hipaconitine and sum of aconite alkaloids per unit weight than those of deeply planted root tubers (control group), while the average weight per root tuber was lower than that of the control group. The total weight of the daughter root tubers per plant of the disbudding group was significantly higher than that of other groups, including the root-pruning group whose average weight per tuber was the highest. In conclusion, disbudding is effective in achieving both higher content of aconite alkaloids and heavier total weight of the root tuber.
    Journal of Natural Medicines 03/2007; 61(2):127-130. · 1.45 Impact Factor
  • Masayuki Mikage, Akane Ochimori
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    ABSTRACT: The Chinese crude drug Qiang-hua was listed as an alias of Du-hua in Shen-nong-ben-cao-jing, an herbal journal written during the Han Dynasty, China. Du-hua and Qiang-hua are recognized as different herbs in China these days; the main botanical origin of Du-hua is Angelica spp. and that of Qiang-hua is Notopterygium spp., of the family Umbelliferae. To make clear the botanical origins of Du-hua and Qiang-hua in ancient China, the authors made a herbological study. The findings were as follows: the name of Qiang-hua was given to the genuine Du-hua, which is produced in Qiang Province, an ancient province located in northwest China; the botanical origin of Qiang-hua is presumed to be Notopterygium incisum Ting ex H.T. Chang and N. forbesii Boissieu based on both the morphology and habitat written in ancient herbal journals. Both species are prescribed as having the plant origin of Qiang-hua in the present Chinese Pharmacopoeia. Therefore, we concluded that Du-hua and Qiang-hua were essentially the same drug, and were originally derived from the Notopterygium species in ancient China.
    Yakushigaku zasshi. The Journal of Japanese history of pharmacy 02/2007; 42(1):17-21.
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    ABSTRACT: Ayurvedaで古くから薬用に供される「Asava」は,生薬を発酵させて製する薬酒である。我が国における一般的な薬酒は生薬を酒に浸漬させてつくられ,これら両者の大きな違いは発酵という過程を経ているか否かという点である。発酵が薬酒の性質にどのように影響を与えるかを明らかにするため,本研究では生姜と大棗を用い,それぞれからAsava製法とチンキ剤製法により薬酒を作製し,揮発性成分比をGC-MSにより測定した。Asava製法については,スリランカにて調査した結果に従った。その結果,Asava製法で作製した薬酒の揮発性成分比は,phenylethanolが30%を占めたのに対し,チンキ剤製法では2%以下と大きく異なっていた。またAsava製法で作製した生姜の薬酒はアルデヒド類(geranial,neral)が1%以下であり,作製中にgeraniol,nerolに還元されたことが示唆された。さらにAsava製法で作製した大棗の薬酒には,zizybeosideのアグリコンであるbenzyl alcoholの含有が認められたことから,アルコール発酵による配糖体の加水分解が示唆された。In Ayurvedic Medicine, some crude drugs and sugar are mixed and fermented, and developed into a herbal liquor called "Asava". During the alcoholic fermentation, the constituents of the crude drug would change chemically. There would thus be a chemical difference between Asava and tincture prepared by soaking the crude drug in alcohol. To investigate the differences between Asava and tincture, we attempted to prepare Asava in our laboratory with ginger rhizomes or jujube fruits and compared the volatile compounds of Asava with those of tincture by GC-MS. The alcohol contents of Ginger Asava and Jujube Asava prepared in the laboratory were 11% and 9.0%, respectively, and were almost the same as Asavas obtained in Sri Lanka. A characteristic difference between the Asava and the tincture was the relative content of phenylethanol (Asava: >30%, tincture: <2%), which is known to be a metabolite of phenylalanine during alcoholic fermentation. In the Ginger Asava, only trace aldehydes such as geranial and neral were found, whereas they were abundant in ginger tincture. Geranial and neral seemed to be reduced to geraniol and nerol. In Jujube Asava, we detected benzyl alcohol that was not detected in jujube tincture. Benzyl alcohol is an aglycon of zizybeoside I or II that was a component of jujube. Therefore, it seemed that glycosides are hydrolyzed by alcoholic fermentation. Hydrolysis of glycosides might help absorption of effective aglycones for the aged.
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    ABSTRACT: Progression of the desertification in northern China has been causing damage to wild Ephedra plants on which we depend for most of supply of the traditional herbal medicine, "Ma huang." The Chinese government encourages the cultivation of Ephedra plants, and Ephedra fields have been reclaimed in the original Ephedra habitats in recent years. We surveyed 7 Ephedra fields that have been recently developed in the Inner Mongolia Autonomous Region and Ningxia Hui Autonomous Region to collect information on Ephedra plant cultivation, especially pertaining to crop species. Specimens taken from those Ephedra fields were genetically and morphologically analyzed, and their ephedrine alkaloid content was examined. DNA analyses of Ephedra specimens, including DNA sequencing of ITS (internal transcribing sequence of nuclear ribosomal DNA) and trn L/F (intron of trnL and intergenic spacer between the trnL and trnF of chloroplast DNA) region and species-specific amplification of trn L/F were conducted to identify Ephedra species. Based on the results of DNA sequencing and morphological determination, the crops grown in 6 fields ware identified as Ephedra sinica, while co-planting of E. sinica and E. intermedia was found in one field where a higher appearance rate of plants with varied morphology from wild Ephedra plants was observed. Furthermore, direct sequencing of the PCR product of the trn L/F region of some specimens from the field and their species-specific PCR showed ambivalent result. Cloning and sequencing of the PCR product of the trn L/F region of those specimens DNA suggested their heteroplasmy, containing both E. sinica- and E. intermedia-type chloroplasts. On the other hand, the profile of the ephedrine alkaloid content was clearly correlated with the result of direct sequencing of the trn L/F region; the specimens showing the E. sinica-type sequence contained more ephedrine than pseudoephedrine, and the specimens of the E. intermedia-type more pseudoephedrine.
    Biological & Pharmaceutical Bulletin 05/2006; 29(4):746-9. · 1.78 Impact Factor
  • Chieko Yoshizawa, Makiko Kitade, Masayuki Mikage
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    ABSTRACT: As we previously reported, ma-huang ([Chinese characters: see text], Ephedrae Herba) has been sometimes used together with mu-zei ([Chinese characters: see text], Equiseti Herba) in medieval China and Japan. We herbologically studied this confusion and found that, in China, the confusion was found in literature in the Song dynasty, and Li Shi-Zhen recorded in Ben-cao-gang-mu that both drugs were morphologically and medicinally the same in the Ming dynasty. Though the main reason why the plant of the genus Equisetum, especially E. ramosissimum Desf., had been substituted for Ephedra plants is thought to be their morphological similarity, the doctors who lived in the area where no Ephedra plants grew might have used Equisetum plants as ma-huang based on Li's description. Owing to this confusion in China, the plants of E. ramosissimum were sometimes imported to Japan as ma-huang, and it caused the tentative use of E. ramosissimum as ma-huang in the middle of the Edo era in Japan.
    Yakushigaku zasshi. The Journal of Japanese history of pharmacy 02/2006; 41(1):9-17.
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    ABSTRACT: The resources of wild Ephedra plants in the Xinjiang Uygur Autonomous Region were surveyed. Ephedra plants mainly grow on the fringes of the Taklimakan Desert and Gureban-tonggute Desert. We found six genotypes of Ephedra przewalskii growing widely in Xinjiang. Three genotypes of Ephedra intermedia were limited to the northern and eastern parts, and Ephedra regeliana scattered in the northern part of Xinjiang. These Ephedra specimens were analyzed for DNA sequences of nuclear ribosomal DNA, internal transcribed spacers 1 and 2, chloroplastic DNA, trnL intron and trnL-trnF intergenic spacer. Intraspecific variation of the nucleotide sequence in E. przewalskii was found in different habitats. Norephedrine, ephedrine, pseudoephedrine, and methylephedrine contents of the specimens were determined. Although Ephedra intermedia of all three genotypes contained ephedrine alkaloids, ephedrine alkaloids were not detected in E. regeliana and E. przewalskii.
    Biological & Pharmaceutical Bulletin 03/2005; 28(2):285-8. · 1.78 Impact Factor
  • Chieko Yoshizawa, Makiko Kitade, Masayuki Mikage
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    ABSTRACT: The botanical origin of a Chinese crude drug Ma-huang in ancient China and the origin of Japanese Ma-huang were herbologically studied. The results showed that the plants of Ephedra sinica Stapf, E. intermedia Schrenk & C. A. Meyer and E. equisetina Bunge were used as Ma-huang in China, and the first species was considered to be of high quality. The characters of Mao-zhou Ma-huang and Tong-zhou Ma-huang printed in Tu-jing-ben-cao, published in the Song Dynasty in China, were identified as E. likiangensis Florin and E. intermedia, respectively, and both species were recognized as excellent Ma-huang in the Ming Dynasty. The word origin of Katsune-kusa, the Japanese name for Ma-huang in the Heian Era, was etymologically considered as meaning the plant having reddish brown roots. In Japan, the plant of Equisetum ramosissimum Desf. var.japonicum Milde, of the family Equisetaceae, was substituted for Ma-huang in the middle of Edo Era, and it was designated that this action was based on the confusion of Ephedra plants and Equisetum plants those days in China.
    Yakushigaku zasshi. The Journal of Japanese history of pharmacy 01/2005; 40(2):107-16.
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    ABSTRACT: Twenty-four Ephedra plants belonging to 8 species grown in the northern and western parts of China were phylogenetically analyzed for their non-coding DNA sequences, internal transcribed spacers (ITSs) of nuclear ribosomal DNA as well as trnL intron and intergenic spacers between trnL and trnF (trnL/ trnF) of the chloroplast. Based on the ITS sequences, the 8 species could be divided into 3 groups: Group 1 (Ephedra intermedia, E. sinica, E. przewalskii), Group 2 (E. equisetina, E. monosperma, E. gerardiana), and Group 3 (E. likiangensis, E. minuta). The species classified into Group 1 grow mainly in the north, Group 3 in the south and Group 2 in the center, suggesting their genetic and geographic relationships. A specific primer set was designed to classify the 3 groups by routine PCR. Combined analysis of ITS and trnL/ trnF differentiated the 8 Ephedra species.
    Planta Medica 12/2004; 70(11):1080-4. · 2.34 Impact Factor
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    ABSTRACT: Based on the Chinese pharmacopeia 2000 ed and Japanese pharmacopeia 14st ed., the original plant differencesof Chinese raditional medicines and Japanese Kampo medicines were compared by making list. The differences and reasons were analyzed.
    Zhongguo Zhong yao za zhi = Zhongguo zhongyao zazhi = China journal of Chinese materia medica 09/2004; 29(8):715-8, 822.
  • Chieko Yoshizawa, Masayuki Mikage, Atsufumi Taru
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    ABSTRACT: The medical commentary "Igaku-tensho-ki," written by Gensaku Manase in the Edo era, Japan, describes the medical treatment of Ho-shun-in, Mrs. Toshiie Maeda, the first seignior of Kaga. The Igaku-tensho-ki is famous for its clinical descriptions of the diseases of many notables such as Emperors and Shoguns by their antonyms, and thus is very important for both medical and historical studies. Several variant versions are known to exist now. As to the description of Ho-shun-in, the details such as the date and place of the treatment, and the recipe of the Kampo medicine are not clear. In this study, to clarify these things, a comparative study of various descriptions in all of the variants was carried out. The results show that the medical treatment was conducted on June 7th of the lunar calendar, in the 11th year of the Keicho Era (1606) in Edo (present-day Tokyo). The disease was thought to be diarrhea caused by the infection of bacteria and round worms. Gensaku , effectively prescribed Sho-ko-en, Kakko-shoki-san with Gorei-san, and Jinryo-byakujutu-san to Ho-shun-in over a 4-day period. All of these prescriptions were basically based on those in "Manbyo-kaishun", archaic medical text written in the Ming Dynasty in China, though they were derived from different origins.
    Yakushigaku zasshi. The Journal of Japanese history of pharmacy 01/2003; 38(1):82-92.
  • Masayuki Mikage, Chieko Yoshizawa
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    ABSTRACT: The Japanese Pharmacopoeia standardizes a Chinese crude drug "Bou-i" as the woody stem of Sinomenium acutum Rherder et Wilson of the family Menispermaceae. The Chinese Pharmacopoeia writes about the same drug under the name of "Fang-ji". The present herbological studies resulted in learning that the original written character for this drug is in China. The ancient Japanese misread it as "--" because of confusion between "--" and "--" in old Chinese herbals. The reason for changing "--" to "--" in China is presumed to be because the pronunciation of these characters changed in the Yuan dynasty, to "--" from "zi" to "si", and to "--" from "ki" to "tsi." That is, the original name "Fang-ji," written in today's Chinese phonetic symbols, can be used without big changes by switching the character from "--" to "--".
    Yakushigaku zasshi. The Journal of Japanese history of pharmacy 02/2002; 37(1):19-27.
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    Kazuhito Kawahara, Masayuki Mikage
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    ABSTRACT: To study the quality of toad venom dried on different metal plates by heating at 105 degrees C, each 20 g sample of fresh toad venom collected in Hei-Long-Jiang Province, China, was dried on (1) brass, (2) copper, (3) glass, (4) acrylic resins, (5) aluminum and (6) stainless-steel, respectively. Twelve bufadienolides, including bufalin and bufotalin, in each sample were then quantitatively analyzed by HPLC. The total levels of bufadienolides in 1000.0 mg of the dried samples were (1) > (2) > (3) > (4) > (5) > (6), varying from 303.44 mg to 420.72 mg. Besides, the color of dried venom became darker in the order of (2), (4), (6), (3), (1) and (5). Though (1) was not in good color, it was superior to the others in chemical quality. These results suggest that it is possible to dry toad venom in short period by heating it at a high temperature on a tray made of brass. This will be a better method for making high quality toad venom than the traditional method. Moreover, the removal of impurities in the fresh venom by the process of filtration through silk succeeded in raising the bufadienolides content significantly.
    Yakugaku zasshi journal of the Pharmaceutical Society of Japan 01/2002; 122(1):117-9. · 0.31 Impact Factor

Publication Stats

91 Citations
38.01 Total Impact Points


  • 2002–2013
    • Kanazawa University
      • • School of Pharmacy
      • • Graduate School of Natural Science and Technology
      Kanazawa, Ishikawa, Japan
  • 2011
    • Peking University
      • School of Pharmaceutical Sciences
      Beijing, Beijing Shi, China
  • 2009
    • University of Toyama
      • Institute of Natural Medicine
      Тояма, Toyama, Japan