The adequacy of the Maoto decoction procedure described in the Shokan-ron text was investigated using the inhibitory activity of blended Ephedra herb on amylase enzyme. This enzyme is closely related to stress, and used clinically as an indicator of stress in humans.α-amylase in isolated mouse plasma was used in the experiment, and the enzymatic activity was measured according to the Caraway method. Two different decoction methods: the general process used today, and that described in the Shokan-ron (Ephedra herb alone is firstly infused, then another 3 crude drugs are added and it is further infused), were compared for yield of the extracts, inhibitory actions of the extracts, their crude polysaccharide fraction (one of the active components), and remaining fractions contained within each extract. The inhibitory activity of the extract was clearly promoted by the latter, ancient decoction method, compared with the former. Hence, the decoction method described in the Shokan-ron is presumed to have the aim of reducing stress to the body caused by Maoto itself, and mitigating the side effects of its component Ephedra herb. This biologically qualitative approach, reflected over the entire decoction process, is different from the simply physicochemically analytical approach which concerns itself merely with particular constituents, such as alkaloids. So, it may be beneficial to scientifically elucidate the significance of this decoction and its ingestion methods, as described by the ancient Shokan-ron and Kinki-yoryaku texts.
Ephedra ; Procedures ; Stress ; Endurance of activity ; Investigations

Since the polyjuice potion ingredient is complex, we need to develop an analysis method with well separation and high stability to perform qualitative analysis. After dividing chemical components of Mahuang Decoction into fat-soluble and water-soluble constituents by gradient extraction, GC-MS was used to analyze the chemical components of the ethyl acetate extraction. The results showed that forty compounds had been identified by NIST MS search 2.0 standard mass spectrometry Library and literatures. Next, UPLC-Q-TOF-MS was applied to idendify the chemical components of the water extraction. The results showed that thirty-nine compounds had been identified by MZmine-2.9.1, Isotope Pattern, fragmentation regularity of mass spectrometry and literatures. This experiment will provide evidences for elucidation of the effective substance in Mahuang decoction and can be used as a simple, shortcut method for analysis and identification for the polyjuice potion.
Chromatography, High Pressure Liquid ; methods ; Ephedra ; chemistry ; Ephedra sinica ; chemistry ; Gas Chromatography-Mass Spectrometry ; methods

BACKGROUND: Skin smoothness in aging skin, in combination of intrinsic aging and photoaging, is of ever-increasing interest which leads to the development of various wrinkle diminuation products including those containing herbal medicines. OBJECTIVES: The purpose of this study was to assess the efficacy of cosmetics containing roots of Ephedra sinica and Betula platyphylla var. japonica on reducing skin furrows. METHOD: Creams containing Ephedra sinica's root and Betula platyphylla var. japonica were applied to forearms of 30 healthy subjects in 2 age groups for 8 weeks and 4 weeks, respectively while silicone skin replicas were taken from medial and lateral sides of the forearm every 4 weeks of the study starting from the week 0. Then the changes of the skin furrows were analyzed with skin visiometer. RESULTS: 1. At week 8, a significant reduction of average roughness and maximum roughness were observed in volunteers younger than age 50 on their Ephedra sinica's root-treated side compared with the placebo-treated side (p<0.05). 2. No statistically significant changes were observed with both Ephedra sinica's root and Betula platyphylla var. japonica in the group of age over 50. 3. Comparison of the changes between treatment groups of both Ephedra sinica's root and Betula platyphylla var. japonica at any assessment time did not show statistically significant differences between the medial and lateral side of forearm, while the treatment group of Ephedra sinica's root showed statistically significant differences between the young and old at week 8 (p<0.05). CONCLUSIONS: The treatment of Ephedra sinica's root seems to be partially effective in improving skin furrows in youth. To our knowledge, this study is the first trial to evaluate the efficacy of both Ephedra sinica's root and Betula platyphylla var. japonica on decreasing wrinkle in the world.
Adolescent ; Aging ; Betula* ; Ephedra sinica* ; Ephedra* ; Forearm ; Humans ; Silicones ; Skin* ; Volunteers

OBJECTIVETo study the karyotype of four Ephedra plants in order to provide the cytologic evidence for the genetic diversity and identification genetic resources of Ephedra.

METHODThe roots of germinating seeds were used to study the karyotype of four Ephedra plants by staining and slide-preparing technique of mitotic chromosomes.

RESULTthe optimal root-sampling time was about 10: 20 - 10:40 am. Using 0.002 mol x L(-1) 8-Hydroxyquinoline to pretreating the intravital root tips, the optimal pretreatment time for E. Sinica, E. intermedina, E. equisetina and E. przewalskii was 4, 5, 4.5 and 3.5 h, respectively. E. przewalskii and E. equisetina were diploid, E. Sinica and E. intermedina were belonged quadruple. The karyotype formulae of the four species were 2n = 2x = 14 = 2M + 8m + 4sm, 2n = 2x = 14 = 10m + 4st, 2n = 4x = 28 = 20m (2SAT) +8st, and 2n = 4x = 28 = 20m (SAT) + 6st + 2sm, respectively.

CONCLUSIONAll the karyotypes of four Ephedra species were 2A type, which was the symmetric karyotype.

Chromosomes, Plant ; genetics ; Ephedra ; cytology ; genetics ; Karyotyping ; methods ; Mitosis

BACKGROUND: In recent years alternative medicine such as herbal medicines has been attractive and useful in dermatologic therapy. But there is little scientific information about its therapeutic effects and potential side effects. OBJECTIVE: We performed this study to assess the whitening effect of the root of Ma Huang. METHODS: Thirty subjects were tanned by UVB irradiation on their abdomen and 4 pigmented patches in square were obtained. In a single-blind fashion, participants applied the root extract of Ma Huang to two pigmented patches and vehicle to another patch on the abdomen twice a day, and applied nothing to the other patch for 8 weeks. Participants were followed up every week and checked on a melanin index with a mexameter. We also checked the last clinical improvement 8 weeks after the application of the root extract of Ma Huang. RESULTS: MED of the middle back was 73+/-24.66cmJ/cm2. Melanin index of the abdomen was 473.2+/-16.27. Eight weeks after application, the mean change percent of the melanin index of the right upper and lower squares applied with the root extract of Ma Huang were +0.02%(p=0.96) and -0.16%(p=0.76). The mean change percent of the left upper square applied with vehicle was -0.08%(p=0.89). The mean change percent of the left lower square untreated was +0.20(p=0.65). One-way layout ANOVA showed no difference among four squares. Clinical evaluation of improvement did not satisfy: no change 36%; slight improvement 53%; moderate improvement 9%; marked improvement 2%. CONCLUSIONS: The root of Ma Huang did not show the whitening effect on the UVB-induced pigmentation checked by the mexameter but showed slight improvement in visual assessment.
Abdomen ; Complementary Therapies ; Ephedra sinica* ; Melanins ; Pigmentation ; Triacetoneamine-N-Oxyl

This study is to establish the characteristic HPLC chromatogram of phenols in Ephedrae Herba, from which to pick out the marker peaks, followed by the analysis of the regularity of their distribution and content in the herbaceous stems of Ephedra sinica, E. intermedia and E. equisetina. The HPLC-DAD method for the characteristic chromatogram as well as quantitative analysis was established. The separation was carried out on a YMC-Pack ODS-A column (4.6 mm x 250 mm, 5 µm), eluted with the mobile phases as 0.01% formic acid aqueous solution (A) and acetonitrile (B) in a linear gradient (0-10 min, 17% B; 10-25 min, 17%-19% B; 25- 33 min, 19%-48% B; 33-35 min, 48%-51% B; 35-44 min, 51% B). The flow rate was kept at 1.0 mL · min⁻¹. The column tem- perature was 40 °C, and the detection wavelength was set at 350 nm (0-16 min) and 330 nm (16-44 min). Forty-six batches of collected samples from three official origins of Ephedrae Herba were detected, whose liquid chromatograms proven to be helpful to the differentiation of different origins. With principal component analysis and the analysis of distribution of peak area, twelve key peaks from the chromatogram were discussed in details on their contributions to the characteristics and differences of three official origins of the herb: peak area of peak 10, 11, 12 were found out to be significantly higher in E. equisetina than in other two origins, whose sum (higher than 146 mAU in E. equisetina) was useful for the discrimination between E. equisetina and the other two origins; peak area of 1 and 4 were respectively higher in E. sinica and E. intermedia than in other official origins, indicating their important effect on the differen- tiation of corresponding origins; peak 8 and 9 were picked out as two characteristic common peaks in three official origins of the herb, whose peak area showed little difference among different origins; further, peak area of other key peaks in the chromatogram also showed some difference among three origins, which make contributions to the differentiation of origins as well. Then, four phenols as 2"-O-α- L-rhamnosyl-isovitexin (1), vitexin (2), pollenitin B (5) and herbacetin-7-O-β-D-glucoside (6) were quantitative analyzed with the above-mentioned method, with good linear relationship and accuracy (recoveries in a range of 97.8%-102.5%). The content of the four phenols were firstly reported in Ephedrae Herba from official origins, which were respectively trace-1.55 (1), trace-0.160 (2), trace-0.284 (5) and trace-0.620 (6) mg · g⁻¹ in all of the tested samples. In addition, the content of these phenols showed differences in three official origins, especially 1, whose content in E. sinica [(0.670 ± 0.88) mg ± g⁻¹] were significantly higher than in other two origins (lower than 0.16 mg ± g⁻¹ besides sample Ei-060630-2-2), and 6, whose average content in E. equisetina [(0.260 ± 0.039 2) mg · g⁻¹] were twice as high as in E. sinica [(0.120 ± 0.270) mg · g⁻¹] and E. intermedia [(0.136 ± 0.485) mg g⁻¹], indicating the important effects of the two constituents on the differentiation among three official origins of the herb. The method established for the characteristic HPLC chromatogram and quantitative analysis of phenols was simple and accurate, and the marker constituents selected may provide new guides for the discrimination of official origins as well as the improvement of quality criteria of EphedraeHerba.
Chromatography, High Pressure Liquid ; methods ; Ephedra ; chemistry ; Phenols ; analysis

OBJECTIVETo optimize the prescription dose of Mahuang decoction in a multi-target manner, in order to provide reference for the quantitative optimization of the prescription dose of the traditional Chinese medicine compound.

METHODThe number of diaphoretic spots in rats, the tracheal antispasmodic rate in guinea pigs and the writhing times by acetic acid in mice were taken as the indexes for evaluating the diaphoretic, antispasmodic and analgesic effects. According to the experimental results of the 16 orthogonal combination prescriptions, a mathematical dose-effect model was built by support vector regression (SVR) and quadratic response surface regression (RSR) respectively. The multi-target optimization was achieved by elitist non-dominated sorting genetic algorithm (NSGA-II) and entropy weight TOPSIS method.

RESULTThe optimal dose of Mahuang decoction after being optimized by SVR modeling contained 17.71 g of Ephedrae Herba, 9.57 g of Cinnamomi Ramulus, 11.75 g of Armeniacae Semen Amarum and 4.39 g of Glycyrrhizae Radix et Rhizoma Praeparata Cum Melle. The optimized result by RSR modeling contained 13.37 g of Ephedrae Herba, 11.61 g of Cinnamomi Ramulus, 11.98 g of Armeniacae Semen Amarum and 5.67 g of Glycyrrhizae Radix et Rhizoma Praeparate Cum Melle. SVR was superior to RSR in both of the forecast capacity and optimization results.

CONCLUSIONSVR-NSGA-II-TOPSIS method could be adopted for the multi-target optimization for the dose of Mahuang decoction and other traditional Chinese medicine compounds. It is proved to be the optimal prescription with the best efficacy, and could provide scientific quantitative basis for determining the dose of traditional Chinese medicine compound prescriptions and developing new traditional Chinese medicines.

Animals ; Cinnamomum ; chemistry ; Drug Compounding ; methods ; Drug Dosage Calculations ; Drug Prescriptions ; Ephedra ; chemistry ; Ephedra sinica ; chemistry ; Glycyrrhiza ; chemistry ; Guinea Pigs ; Mice ; Rats

Ma-huang (Ephedra sinica) is a herb that exhibits sympathomimetic activity. We report a patient with acquired myopathy following ingestion of a Ma-huang-based herbal remedy for 6 months. A 50-year-old woman was admitted for progressive muscular weakness. An electrodiagnostic study and muscle biopsy sampling revealed generalized active myopathy with nonspecific inflammation. It appears that Ma-huang, along with Raynaud's phenomenon and pericardial effusion, may be a causative agent of myopathy. As such, attention should be paid to the possibility that herbal medication with Ma-huang can result in myopathy.
Biopsy ; Eating ; Ephedra ; Female ; Humans ; Inflammation ; Middle Aged ; Muscle Weakness ; Muscles ; Muscular Diseases ; Pericardial Effusion

OBJECTIVETo identify Ephedrae Herba using the ITS2 barcode and to secure its quality and safety in medication.

METHODTotal genomic DNA was isolated from Ephedrae Herba and its closely related species. Nuclear DNA ITS2 sequences were amplified, and purified PCR products were sequenced. Sequence assembly and consensus sequence generation were performed using the CodonCode Aligner. The Kimura 2-Parameter (K2P) distances were calculated using software MEGA 5.0. Identification analyses were performed using BLAST1, Nearest Distance and neighbor-joining (NJ) methods.

RESULTThe intra-specific genetic distances of Ephedrae Herba were ranged from 0 to 0.002. The inter-specific genetic distances between Ephedrae Herba and its closely related species were ranged from 0.004 to 0.034. All the three methods showed that ITS2 could discriminate Ephedrae Herba from its closely related species correctly.

CONCLUSIONThe ITS2 region is suitable to be used for authentication of Ephedrae Herba, and our study further confirmed the effectiveness of ITS2 to identify traditional Chinese medicinal materials.

DNA Barcoding, Taxonomic ; methods ; DNA, Ribosomal Spacer ; genetics ; Ephedra sinica ; classification ; genetics

OBJECTIVETo investigate the change regularity of ephedrine and glyrihhzine acid in Ephedra sinila and Glycyrrhiza uralencis pair medicines and in Maxing Shigan decoction.

METHODThe contents of ephedrine and glycyrrhizic acid were determined by HPLC in samples of E. sinica extracts, G. uralencis extracts, pair medicines extracts of Maxing Shigao decoction sinica and G. uralencis, and extracts of E. sinica.

RESULTThere were no significant difference in ephedrine contents amoung different samples; the contents of glycyrrhizic acid were lower in decoctions of Maxing Shigan decoction than in G. uralencis decoction and pair medicines extracts.

CONCLUSIONMacromolecular complex was NOT formed by ephedrine and Glycyrrhizic acid in decoctions containing pair medicines of E. sinica and G. uralencis.

Chemistry, Pharmaceutical ; Drugs, Chinese Herbal ; chemistry ; Ephedra sinica ; chemistry ; Ephedrine ; chemistry ; Glycyrrhiza ; chemistry ; Glycyrrhizic Acid ; chemistry