1.To Get the Resources of Crude Drugs
Hiroyuki TOMITSUKA ; Katsuko KOMATSU
Kampo Medicine 2010;61(5):754-773
2.Authentication of Curcuma species (Zingiberaceae) based on nuclear 18S rDNA and plastid trnK sequences.
Hui CAO ; Yohei SASAKI ; Hirotoshi FUSHIMI ; Katsuko KOMATSU
Acta Pharmaceutica Sinica 2010;45(7):926-33
Curcuma drugs have been used discriminatingly for invigorating blood circulation, promoting digestion, and as a cholagogic in China. However, there is confusion about the drug's botanical origins and clinical uses because of morphological similarity of Curcuma plants and drugs. Comparative sequencing of the 18S rRNA gene in nuclear ribosomal DNA (rDNA) and trnK gene in chloroplast DNA (cpDNA) was carried out in order to examine interspecies phylogeny and to identify ultimately Curcuma species. A total of a hundred of accessions of eighteen species were analyzed. This resulted in an aligned matrix of 1810 bp for 18S rDNA and 2 800 bp for trnK. 18S rDNA sequence divergence within the ingroup ranged from 0-0.05%, trnK ranged from 0-0.19%. One base transversion-substituted site (from cytosine to thymine) was observed from the upstream of 18S rDNA at nucleotide position 234 in C. kwangsiensis and Japanese population of C. zedoaria which have separated genetic distance to other Curcuma taxa. Two noncoding regions embedded in trnK intron showed higher variability, including nucleotide substitutions, repeat insertion and deletions. Based on consensus of relationship, eighteen major lineages within Curcuma are recognized at the species level. The results suggest that Curcuma is monophyletic with 100% bootstrap support and sister to the genera Hedychium and Zingiber. The trnK sequences showed considerable variations between Curcuma species and thus were revealed as a promising candidate for barcoding of Curcuma species, which provide valuable characters for inferring relationship within species but are insufficient to resolve relationships among closely related taxa.
3.Questionnaire Assessment of Training in the Use of Kampo Medicine in an Outpatient Clinic.
Takashi ITOH ; Toshiaki KITA ; Naotoshi SHIBAHARA ; Hirotoshi FUSHIMI ; Yutaka SHIMADA ; Katsuko KOMATSU ; Katsutoshi TERASAWA
Medical Education 2001;32(1):31-38
We used questionnaires to assess training in the use of Kampo medicine which took place mostly at our outpatient clinic. Most of the 18 trainees (6 medical doctors and 12 medical students) felt that the quantity and the quality of the lectures and the amount of time spent in the outpatient clinic were satisfactory. In addition, all trainees thought that attending the outpatient clinic was the most instructive activity and made the deepest impression, but 14 of the 18 trainees (78%) thought that it was a difficult learning strategy. Fifteen trainees (83%) rated this training curriculum as good or very good. Sixteen trainees (88%) were conscious of a change in their study or work attitudes 3 to 6 months after training; this result suggests that this curriculum has an educational effect.
4.Molecular identification of six medicinal Curcuma plants produced in Sichuan: evidence from plastid trnK gene sequences.
Acta Pharmaceutica Sinica 2003;38(11):871-875
AIMTo establish a rapid and simple molecular identification method for six medicinals: Curcuma: C. longa, C. phaeocaulis, C. sichuanensis, C. chuanyujin, C. chuanhuangjiang, and C. chuanezhu in Sichuan Province.
METHODSA molecular approach (trnK nucleotide sequencing) was used in this study.
RESULTSThe sequenced entire chloroplast trnK gene region spanned 2,699-2,705 bp. The matK gene (an intron embodied in trnK gene) sequence and the intron spacer region of the trnK gene have great diversity within these six medicinal Curcuma species. There were six single bases substitutions between trnK coding region and matK region, the 9-bp deletion and 4-bp or 14-bp insertion repeat at some sites of matK region in each taxon.
CONCLUSIONThese relatively variable sequences were potentially informative in the identification for these six Curcuma species at the DNA level.
Base Sequence ; Chloroplasts ; genetics ; Curcuma ; classification ; genetics ; DNA Mutational Analysis ; methods ; DNA, Plant ; genetics ; Genes, Plant ; Introns ; Phylogeny ; Plants, Medicinal ; classification ; genetics ; RNA, Transfer, Lys ; genetics ; Sequence Analysis, DNA ; methods ; Sequence Deletion
5.A novel approach to quality evaluation of root of Scutellaria baicalensis by DPPH free radical scavenging.
Yu-ping LIU ; Purusotam BASNET ; Katsuko KOMATSU ; Hui CAO
China Journal of Chinese Materia Medica 2002;27(8):575-619
OBJECTIVETo develop a simple, reliable approach for evaluating the quality of Huangqin (Scutellaria baicalensis).
METHODTo determine the DPPH free radical scavenging activity and assay of four bioactive components: baicalin, baicalein, wogonin and wogonin-7-O-glucuronide by HPLC.
RESULTThe correlative relationship between DPPH free radical scavenging activity and baicalin content was obtained.
CONCLUSIONBioassay of DPPH free radical scavenging activity could be used as one of the methods for quality evaluation of Chinese drug Huangqin.
Biphenyl Compounds ; Flavanones ; Flavonoids ; chemistry ; isolation & purification ; pharmacology ; Free Radical Scavengers ; pharmacology ; Free Radicals ; Molecular Structure ; Picrates ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Scutellaria baicalensis ; chemistry
6.Authentication of Curcuma species (Zingiberaceae) based on nuclear 18S rDNA and plastid trnK sequences.
Hui CAO ; Yohei SASAKI ; Hirotoshi FUSHIMI ; Katsuko KOMATSU
Acta Pharmaceutica Sinica 2010;45(7):926-933
Curcuma drugs have been used discriminatingly for invigorating blood circulation, promoting digestion, and as a cholagogic in China. However, there is confusion about the drug's botanical origins and clinical uses because of morphological similarity of Curcuma plants and drugs. Comparative sequencing of the 18S rRNA gene in nuclear ribosomal DNA (rDNA) and trnK gene in chloroplast DNA (cpDNA) was carried out in order to examine interspecies phylogeny and to identify ultimately Curcuma species. A total of a hundred of accessions of eighteen species were analyzed. This resulted in an aligned matrix of 1810 bp for 18S rDNA and 2 800 bp for trnK. 18S rDNA sequence divergence within the ingroup ranged from 0-0.05%, trnK ranged from 0-0.19%. One base transversion-substituted site (from cytosine to thymine) was observed from the upstream of 18S rDNA at nucleotide position 234 in C. kwangsiensis and Japanese population of C. zedoaria which have separated genetic distance to other Curcuma taxa. Two noncoding regions embedded in trnK intron showed higher variability, including nucleotide substitutions, repeat insertion and deletions. Based on consensus of relationship, eighteen major lineages within Curcuma are recognized at the species level. The results suggest that Curcuma is monophyletic with 100% bootstrap support and sister to the genera Hedychium and Zingiber. The trnK sequences showed considerable variations between Curcuma species and thus were revealed as a promising candidate for barcoding of Curcuma species, which provide valuable characters for inferring relationship within species but are insufficient to resolve relationships among closely related taxa.
China
;
Curcuma
;
classification
;
genetics
;
DNA Mutational Analysis
;
DNA, Chloroplast
;
genetics
;
DNA, Plant
;
genetics
;
Introns
;
Japan
;
Molecular Sequence Data
;
Nucleic Acid Amplification Techniques
;
Phylogeny
;
Plants, Medicinal
;
classification
;
genetics
;
Plastids
;
genetics
;
RNA, Ribosomal, 18S
;
genetics
;
Sequence Analysis, DNA
7.Research of ginsenosides in kou zi qi using HPLC-MS-MS.
Kun ZOU ; Zhao-xia LIU ; Shu ZHU ; Shao-qing CAI ; Katsuko KOMATSU
Acta Pharmaceutica Sinica 2004;39(5):385-388
AIMTo analyze the ginsenosides in Kou zi qi (rhizomes of Panax japonicus C. A. Mey. var. major (Burkill) C. Y. Wu et K. M. Feng), and to supply evidences for chemotaxanology of Panax species and clinical uses of Kou zi qi.
METHODSThe ginsenosides were isolated by HPLC, then the positive- and negative-ion API-MS/MS of constituents collected from HPLC were measured.
RESULTSEight ginsenosides were identified as ginsenoside Re, ginsenoside Ro, chikuseksusaponins IV, IVa, notoginsenoside R2, ginsenosides Rb1, Rc and Rd, respectively, based on comparison of retention time with those of standards by HPLC, and analysis on their API-MS/MS data. Ginsenoside Ro and chikuseksusaponin IVa are the major components of Kou zi qi.
CONCLUSIONThis plant had a close relationship to P. stipuleanatus, P. zinginensis and P. japonicus var major; a relatively remote relationship to P. ginseng and P. quinquefolius, in a view of chemotaxanology. Ginsenoside Ro and chikuseksusaponin IVa might be the anti-inflammatory constituents of Kou zi qi.
Chromatography, High Pressure Liquid ; Ginsenosides ; isolation & purification ; Panax ; chemistry ; Phylogeny ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Spectrometry, Mass, Electrospray Ionization
8.Effect of total triterpenoids of Chaenomeles speciosa on PPARγ/SIRT1/NF-κBp65 signaling pathway and intestinal mucosal barrier of ulcerative colitis induced by DSS in mice.
Xing-Jun XIONG ; Xiao-Mei LI ; Yu-Min HE ; Xiao-Qin LI ; Hai-Yan XU ; Min-Lu FENG ; Hai-Bo HE ; Ji-Hong ZHANG ; Shu ZHU ; KOMATSU KATSUKO ; Kun ZOU
China Journal of Chinese Materia Medica 2018;43(21):4295-4304
To observe the effect of total triterpenoids of Chaenomeles speciosa on PPARγ/SIRT1/NF-κBp65 signaling pathway and intestinal mucosal barrier of ulcerative colitis induced by dextran sulfate sodium (DSS) in mice, C57BL/6 mice were randomly divided into normal group, model group, total triterpenoids of C. speciosa (50, 100 mg·kg⁻¹) groups and sulfasalazine (250 mg·kg⁻¹) group. The ulcerative colitis (UC) model was induced by orally administering 2.5% DSS to the experimental mice, and the corresponding drugs were given to each group 3 days before the administration with 2.5% DSS. The normal group and the model group were given the equal volume of 0.5% carboxymethyl cellulose sodium solution by gavage continuously for 10 days, q.d. The general conditions of the mice were observed on a daily basis, and the disease activity index (DAI) score was recorded. On the 10th day after the treatment, mice were put to death, the contents of TNF-α, IL-1β, IL-6, IFN-γ, IL-4 and IL-10 in the blood were detected, colon length was measured, colon mucosa damage index (CMDI) score was calculated, and MPO activity detection and histomorphology analysis were conducted. Real-time PCR was applied to detect the mRNA expressions of E-cadherin, occluding,MUC2 and TFF3; the protein expressions of SIRT1, IKKβ, p-IKKβ, IκBα, p-IκBα and cytosol and nucleus PPARγ, NF-κBp65 in intestinal tissue were detected by western blot. The results indicated that total triterpenoids of C. speciosa (50, 100 mg·kg⁻¹) could significantly improve the general conditions of UC mice, reduce the DAI, CMDI and histopathological scores, increase the colon length, reduce the colonic mucosa ulcers, erosion and inflammatory infiltration, restore the normal intestinal mucosal barrier function, reduce the contents of TNF-α, IL-1β, IL-6, IFN-γ, increase the contents of IL-4 and IL-10 in the blood, inhibit MPO activity in colon tissue, up-regulate the mRNA expressions of E-cadherin, occludin, MUC2 and TFF3 in colon tissue, down-regulate the protein expressions of cytosol PPARγ, tissue p-IKKβ, p-IκBα and nucleus NF-κBp65 in the colon tissue, decrease the p-IKKβ/IKKβ and p-IκBα/IκBα ratios, up-regulate the protein expressions of nucleus PPARγ, tissue SIRT1 and cytosol NF-κBp65 (<0.05 or <0.01, respectively), with a dose-effect relationship between the total triterpenoids of C. speciosa treated groups. These findings suggested that total triterpenoids of C. speciosa had a significantly therapeutic effect on UC mice induced by DSS, its mechanism might be related to the regulation of PPARγ/SIRT1/NF-κBp65 signaling pathway, the inhibition of pro-inflammatory factor formation and the up-regulation of protein expression of protective factors.
Animals
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Colitis, Ulcerative
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chemically induced
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drug therapy
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Colon
;
drug effects
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Dextran Sulfate
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Disease Models, Animal
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Intestinal Mucosa
;
drug effects
;
Mice
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Mice, Inbred C57BL
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PPAR gamma
;
metabolism
;
Random Allocation
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Rosaceae
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chemistry
;
Signal Transduction
;
drug effects
;
Sirtuin 1
;
metabolism
;
Transcription Factor RelA
;
metabolism