Study of antidiabetic and antioxidant activities for 2 species of birch leaves / Монголын эм зүй, эм судлал

Abstract@#The birch leaves were used as a substitute for birch bark, buds and chaga of birch in traditional medicine because the birch leaves are considered to be less toxic. Numerous researches conducted in Russia, Bulgaria, Japan, and China on <i>B.pubescens, B. pendula, B.Rezniczenkoana (Litv) </i> Schischk, <i>B.humilis</i> Schrank, and <i>B.mandshurica</i> Rgl Nakai found that birch barks and leaves contain antioxidants and they have anti-cancer, anti-yeast, antibacterial, anti-inflammatory, liver protective and bile secretion induction properties. The studies conducted on animals with diseases showed that the birch leaves had anti-inflammatory properties on the gastric mucosa during acute stress, as well as anti-biliary and giardiasis. The birch leaf phytopreparations experimentations used on animals showed reduced peripheral tissue insulin resistance and lowered blood sugar. Mongolian traditional medicinal journals noted that the birch barks are used to treat inflammatory acute diseases. Therefore, this study was performed to determine the effects of two species of birch leaves on blood sugar and antioxidant activities in diabetes-induced rats.@*The study materials and methods@#The study was conducted in the Pharmacology Research Laboratory of the Monos Group’s Institute of Pharmacology. 40 WISTAR, non-linear white rats weighing 150-204 g were used in the experiments. Dry extract of birch leaves of the two species (Alloxan monohydrate Tokyo Chemical Industry LTD), IGM-100 3A blood glucose meter (Blood glucose test meter, Infopia LTD, Brussels Belgium) and sugar test (Blood glucose test strip only, province, China) were used for the experiment. Lenzen’s (2008) method was used to induce Alloxan diabetes in the rats and the antioxidant properties were determined by the antioxidant activity kit (Rat Malondialchehyche Elisa KIT, cat. № EKRAT- 0266, Jilin).@*Study Result@#The blood glucose level of the control group with diabetes lowered from 31.5 mmol/l to 17.1 mmol/l in 14 days. As for the <i>B.platyphylla</i> Sukacz group, the blood glucose level reduced to 6.3 mmol/l and the <i>B.hippolytii. </i> Sukacz group’s blood glucose level reduced to 6.9 mmol/l in 14 days.</br> The study results showed that <i>B.hippolytii </i>Sukacz birch leaves and <i>B.platyphilla</i> Sukacz birch leaves’ extracts reduced the maximum level of MDA dilution (4.8 nmol/ml) of B.hippolytii Sukacz and B.platyphilla Sukacz groups by 33.9% and 53.5% respectively. This suggests that the birch leaves had antioxidant effect.@*Conclusion@#<i>B.hippolytii </i>Sukacz birch leaves and <i>B. platyphilla </i> (Sukacz) birch leaves lowered the blood glucose level and had antioxidant properties on diabetes.

Toxicology study of Hippolytii birch (B. Hippolytii. Sukacz) leaves and flat leaved birch (B.Platyphylla. Sukacz) / Монголын эм зүй, эм судлал

Abstract@#Numerous researches conducted in Russia, Bulgaria, Japan, and China on <i>B.pubescens, B. pendula, B.rezniczenkoana (Litv) </i> Schischk, <i>B.humilis</i> Schrank, <i>B.mandshurica</i> Rgl Nakai found that birch barks and leaves contain antioxidants and they have anti-cancer, anti-fungi, antibac- terial and anti-inflammatory properties, protect liver and promote bile secretion. Flat leaved birch (<i>B.platyphylla</i> Sukacz) cortex contains betulin and lupeol of triterpenoids and it’s leaves contain flavonoid and polyphenol compounds. The amounts of compounds found in the cortex are smaller than leaves. Specifically, the amount of flavonoid in leaves is more contained than the that of cortex and leaf buds. In any pharmacology study of new medicines, determination and evaluation of toxicity is the first priority. According to scientific evidences that birch leaves are considered to have less toxins. Not many studies have been conducted on determining toxicity of birch leaves in Mongolia. Therefore, the purpose of this research is to study the species of birches, hippolytii birch (<i>B.hippolytii. </i> Sukacz) and flat leaved birch (<i>B.platyphylla. </i> Sukacz), that were noted to have medical properties in traditional medications and identify their acute toxicity using dry extract and determine mortality dosage (LD₅₀) on animals.@*Research materials and methods@#Evaluation of the acute toxicity of birch leaves was conducted in Pharmacology laboratory of Monos group’s Drug Research Institute between June 19, 2020 and August 10. In this research, 150-204 g of WISTAR breed non-linear 44 white rats were used and 20 g of <i>B.Hippolytii’</i>s dry extract and 20 g of B. <i>Platyphylla</i> ‘s dry extract were injected.</br> The experiments to determine the toxicity of dry extracts of <i>B. hippolytii</i> and <i>B. platyphylla</i> (LD₅₀) were conducted according to Litchfield and Wilcoxon’s method and subcutaneous injects were per formed in the pelvic area of the rats. @*Results of determining acute toxicity level@#The experiments to determine the acute toxicity level of the birch’s dry extracts followed Litchfield and Wilcoxon’s method with 2-stage. LD₅₀ level was determined from the first stage of the research using G.N.Pirshen’s method and the toxicity level was identified using K.K.Sidorov’s toxicity categorization.</br> From the acute toxicity research, no-observed-adverse-effect level (NOAEL), animal daily dosage and human daily dosage (experimental) were determined. LD₅₀ 2950 mg/kg was determined as a result of acute toxicity research of B.hippolytii and B.platyphilla leaves’ dry extract.

Chemical constituents from barks of Betula platyphylla / 中草药

Objective: To study the chemical constituents from the barks of Betula platyphylla. Methods: The chemical constituents were isolated by silica gel column chromatography and HPLC, and its structure were identified by their spectral data and physicochemical properties analysis. Results: A total of 20 compounds were isolated from B. platyphylla, which were identified as 3-oxo-24-methylenecycloartan (1), sitosterone (2), 3β-acetyl oleanolic acid (3), lupeol (4), citrostadienol (5), betulinaldehyde (6), 3β-acetoxyl-11α,12α-epoxy-oleanan-13β,28-olide (7), oleanolic acid (8), erythrodiol (9), 3-hydroxy-1,7-bis(4',4″- dihydroxyphenyl)- heptane (10), β-sitosterol (11), 3β,20-dihydroxylupane (12), 3β-O-caffeoyl-betulin (13), platanic acid (14), 3β,12α-dihydroxyoleanan- 13β,28-olide (15), 30-norlupane-3β,28-diol-20-one (16), betulinic acid (17), betulin (18), 1,7-bis (4',4″-dihydroxyphenyl)-hept- 4-ene-3-one (19), and 5-hydroxy-1,7-bis (4',4″-dihydroxyphenyl)-hept-3-one (20). Conclusion: Compounds 1, 5, 7, 12, 14-16 are isolated from B. platyphylla for the first time, and compounds 1, 5, 7, 14-16 are isolated from betula genus for the first time.

Effect of carbon monoxide on growth and triterpenoid production in Betula platyphylla suspension cells / 中草药

Objective: To analyze the effect of carbon monoxide (CO) on the growth of Betula platyphylla suspension cells and triterpenoid production. Methods: CO donor hematin of 5, 10, 20, and 30 μmol/L were added to eight-day-old B. platyphylla suspension cells, and the change of triterpenoid content and gene expression of FPPS, LUS, CAS1, CAS2, and β-AS related to the triterpenoid synthesis were analyzed by chemical colorimetry, high performance liquid chromatography and real-time PCR, respectively. Results: CO treatment did not have a significant effect on dry weight of B. platyphylla suspension cells except for 2-8 d at 20 μmol/L and 30 μmol/L CO treatment. CO treatment also did not have a significant effect on pH value and malondialdehyde content except for 30 μmol/L CO treatment. CO treatment significantly increased the content of triterpenoid, betulin and oleanolic acid. the content of triterpenoid, betulin and oleanolic acid were the highest after 1 d at 30 μmol/L CO treatment, 4 d at 10 μmol/L CO treatment and 2 d at 10 μmol/L CO treatment, which was 1.3, 1.5, and 4.2 times of the control, respectively. Triterpenoid production in B. platyphylla suspension cells induced by CO was further confirmed by key enzyme genes of FPPS, LUS, CAS1, CAS2, and β-AS related to the triterpenoid synthesis by RT-PCR. Conclusion: CO treatment could effectively enhance the triterpenoid production in B. platyphylla suspension cell culture.

The Cytotoxic Constituents of Betula platyphylla and their Effects on Human Lung A549 Cancer Cells

During the screening for cytotoxic compounds from plants grown in Korea, Betula platyphylla (BP) showed potent activity against the adenocarcinomic human alveolar basal epithelial A549 cell line. To identify the cytotoxic components from BP, the CH₂Cl₂ fraction with the most significant cytotoxic effect was applied to the column chromatographies. Seven compounds were isolated: lupeol (1), betulinic acid (2), (−)-rhododendrol (3), platyphyllenone (4), platyphyllone (5), (−)-centrolobol (6), and oleanolic acid (7). Among them, three diarylheptanoids (4 – 6) exhibited cytotoxicity toward A549 cells. Especially, 50 µM of 4 reduced A549 cell viability to 18.93 ± 0.82% compared to control (100.00 ± 21.48%). Lactate dehydrogenase (LDH) leakage and intracellular reactive oxygen species (ROS) production were also induced by 50 µM 4. This is the first report on the cytotoxic effect of BP-derived diarylheptanoids 4–6 against A549 cells. The compound 4 may be useful for the development of early hit compounds for non-small cell lung carcinoma, but the consideration about selectivity of 4 is required since 4 also showed the cytotoxicity in the human normal lung epithelial BEAS-2B cell line.

LC/MS-based Analysis of Bioactive Compounds from the Bark of Betula platyphylla var. japonica and Their Effects on Regulation of Adipocyte and Osteoblast Differentiation

Betula platyphylla var. japonica (Betulaceae), also known as Asian white birch, is an endemic medicinal tree, the bark of which has been used in Chinese traditional medicine for the treatment of various inflammatory diseases. In our continuing search for bioactive compounds from Korean natural resources, a phytochemical investigation of the bark of B. platyphylla var. japonica led to the isolation of 7-oxo-β-sitosterol (1) and soyacerebroside I (2) from its ethanol extract as main components by liquid chromatography (LC)/mass spectrometry (MS)-based analysis. The structures of isolates were identified by comparison of ¹H and ¹³C nuclear magnetic resonance spectroscopic data and physical data with the previously reported values and LC/MS analyses. To the best of our knowledge, this is the first study to demonstrate that the isolated compounds, 7-oxo-β-sitosterol and soyacerebroside I, were isolated in B. platyphylla var. japonica. We examined the effects of the isolates on the regulation of adipocytes and osteoblast differentiation. These isolates (1 and 2) produced fewer lipid droplets compared to the untreated negative control in Oil Red O staining of the mouse mesenchymal stem cell line without altering the amount of alkaline phosphatase staining. The results demonstrated that both compounds showed marginal inhibitory effects on adipocyte differentiation but did not affect osteoblast differentiation.

Cloning of BpTRX gene and promoter and expression pattern analysis under H2S treatment in Betula platyphylla / 中草药

Objective To study the full-length, promoter sequences and its coding structure and properties of thioredoxin (Trx) gene of Betula platyphylla (BpTRX), and reveal the expression pattern of BpTRX under H2S treatment. Methods The BpTRX gene was cloned by PCR, and the promoter region sequences of BpTRX was obtained by using chromosome walking technique. The BpTRX gene, the promoter and its encoded protein were analyzed by bioinformatics software. The phylogenetic tree of BpTRX was constructed. The expression patterns of BpTRX gene under H2S exogenous stress were analyzed by quantitative real-time PCR. Results The full-length of BpTRX gene is 351 bp, encoding 117 amino acids. The BpTRX gene was closely related to the TRX-H protein of castor bean, soybean, alfalfa and grape. The obtained BpTRX promoter region sequences are 873 bp, which contains the essential elements of transcription and a large number of stress response and hormone response elements. Conclusion The full length and partial promoter sequences of the BpTRX gene were obtained. The response trend of BpTRX gene to H2S treatment was in the form of bimodal, which was first rises, then decreases, then rises and then declines.

Analysis on structure and expression pattern of BpHMGR gene in Betula platyphylla / 中草药

Objective: To study the full-length gene structure characteristics and coding protein properties of 3-hydroxy-3-methyl-glutaryl-CoA reductase (BpHMGR) from Betula platyphylla (birch). This reveals the mechanism of rhythm expression pattern and its signaling molecule in response to exogenous NO. Methods: The BpHMGR gene in birch was cloned by RACE technology. BpHMGR and its encoded protein were analyzed in detail by bioinformatics software. And the expression pattern in the circadian variation of BpHMGR gene was analyzed by real-time PCR. Results: The full-length of BpHMGR gene was 1764 bp, contained the complete ORF, encoding 587 amino acids (Genebank ID: KJ197336). According to BpHMGR amino acid sequence, phylogenetic tree was constructed. And the effect of circadian rhythm and exogenous NO on the transcription level of BpHMGR in the leaves of B. platyphylla was investigated. Conclusion: BpHMGR gene, soybean (XP_003519474.1), grape (XP_002275827.1), and alfalfa (XP_003617066.1) have close genetic relationship. BpHMGR changes with the diurnal cycle showing the expression characteristics of rhythm, and reaches the peak at 9 pm. The up-regulation expression of key enzyme gene HMGR in the exogenous NO-induced birch intracellular triterpenoids metabolic pathway is initially revealed, so as to promote the synthesis of triterpene oleanolic acid product.

Diacylglycerol Compounds from Barks of Betula platyphylla with Inhibitory Activity against Acyltransferase / 中草药·英文版

Objective: To identify the active compounds from the barks of Betula platyphylla for inhibitory on diacylglycerol acyltransferase (DGAT1). Methods: Bioassay-guided fractionation resulted in the isolation of DGAT1 inhibitory activity of lupane triterpenes. Results: Ten compounds were identified as lupenone (1), lupeol (2), betulinic acid (3), betulinaldehyde (4), betulin (5), 3-deoxybetulonic acid (6), glochidonol (7), lup-20/29-ene-1β/3β-diol (8), 3α-hydroxy-lup-20(29)-en-23,28-dioic acid (9), and 3α,11α-dihydroxy-23-oxo-lup-20(29)-en-28-oic acid (10). Compounds 3-6, 9, and 10 inhibited DGAT1 with IC50 values ranging from (11.2 ± 0.3) to (38.6 ± 1.2) μmol/L. Conclusion: Compounds 6, 9, and 10 are first isolated from the barks of B. platyphylla., and compounds 3-6, 9, and 10 from the barks of B. platyphylla are responsible for the inhibition on DGAT1. © 2014 Tianjin Press of Chinese Herbal Medicines.

Bioinformatics and expression pattern analysis of BpNOS gene in Betula platyphylla / 中草药

Objective: Nitric oxide synthase (NOS) is a kind of important NOS in organisms. The gene cloning and expression pattern analysis of NOS in plant laid the foundation for study on the biological functions of nitric oxide in plants. Methods: NOS gene in B. platyphylla was cloned by RACE technology, named BpNOS. Bioinformatics and molecular evolution analysis, abiotic stress, and signal induced of the gene were preliminarily carried out. Results: This full-length gene was 1 806 bp with the complete ORF, encoding 601 amino acids (Genebank ID: KJ197336). BpNOS was an unstable hydrophobic protein. This protein may exist signal peptide with transmembrane ability. The alpha helix, extension chain, random coil distributed throughout the protein. Molecular evolution analysis results showed that the genetic distance of BpNOS gene was closer to the gene genetic distance of grape species NOS, which indicates the relatively close relationship between them; While the genetic distance of BpNOS gene was farther from soybean and alfalfa, which explains that the relationship of them is far. The expression of BpNOS has rhythm, reached the maximum at 15: 00, 12 times as at 0: 00. The expression of BpNOS gene could be inhibited by salt, low temperature, and heavy metal cadmium stress, but in a different response pattern. The expression of BpNOS gene could be promoted by salicylic acid and exogenous NO. Conclusion: Abiotic stress could inhibit the activity of NOS. The salicylic acid and exogenous NO could be synthesized by NOS pathway to regulate endogenous NO. This paper lays a solid foundation for the further research on the metabolic regulation of NO in B. platyphylla.

Effect of putrescine on sucrose metabolism and triterpenoid accumulation in Betula platyphylla suspension cells / 中草药

Objective: To analyze the effect of exogenous putrescine on triterpenoid accumulation and sucrose metabolism. Methods: Putrescines (0.1, 1, and 5 mmol/L) were added to the eight-day-old suspension cell culture, and the content changes of sucrose, fructose, glucose, and triterpenoid, the activity of sucrose phosphate synthase (SPS) and sucrose synthase (SS) were analyzed by colorimetry. Results: Putrescine increased the content of sucrose in cells but decreased the conent in culture medium. Among them, the content of sucrose was most enhanced after putrescine induction for 6 h; The increasing rate was 169.41%. However, the contents of fructose and glucose were only increased in 6 h under the putrescine treatment, the other treatments decreased the contents of fructose and glucose. In addition to 5 mmol/L putrescine treatment, putrescine enhanced the activities of SPS and SS. The content of triterpenoid in suspension cells was increased in 12 h after 5-40 g/L sucrose induction, and the content of triterpenoid was most induced by 20 g/L sucrose, the increasing rate was 52.16%. In addition to increasing 27.37% on triterpenoid content in cells treated by 1 mmol/L putrescine and 5 g/L sucrose, the treatment of 1 mmol/L putrescine and other sucrose concentration decreases the accumulation of triterpenoid. Conclusion: Putrescine could increase the sucrose content and triterpenoid accumulation in suspension cell culture, and we preliminarily inferred that the induction of sucrose by putrescine could participate in the accumulation of triterpenoid in birch cells.

Polyamine-mediated triterpenoid synthesis in suspension cells of Betula platyphylla induced by fungal elicitor / 中草药

Objective: To clarify whether polyamine-mediated triterpenoid synthesis in birch (Betula platyphylla) suspension cells induced by fungal elicitor. Methods: Fungal elicitor (40 μg/mL), putrescine (Put, 1 mmol/L), and D-arginine (D-Arg, 2 mmol/L) were added to the eight-day-old suspension cell culture, the content changes of triterpenoid and free polyamines were analyzed by chemical colorimetry and HPLC. The effect of polyamine on triterpenoid synthesis in B. platyphylla suspension cells induced by fungal elicitor was analyzed by pharmacology and restoration experiment. Results: After the treatment of fungal elicitor or Put, the contents and yields of free polyamines and triterpenoid were increased. Among of them, the triterpenoid content was the highest after 24 h treatment, the increasing rates were 68.54% and 30.34%, respectively. The triterpenoid content was increased by the cotreatment of fungal elicitor and Put, but the increasing degree of yield was lower than that by the treatment of fungal elicitor alone. Compared with the fungal elicitor alone, the cotreatment of fungal elicitor and D-Arg decreased the triterpenoid content by 40.57% in the highest degree of decreasing after 24 h treatment. In restoration experiment, with the treatment time prolonging, the effect of fungal elicitor, Put, or cotreatment of fungal elicitor and D-Arg on triterpenoid synthesis was decreasing to the control level finally. Conclusion: Polyamine could mediate the triterpenoid synthesis in cells of B. platyphylla induced by fungal elicitor.

Effect of polyamine on growth of Betula platyphylla suspension cells and triterpenoid accumulation / 中草药

Objective: To analyze the effect of exogenous polyamine on the growth of Betula platyphylla suspension cells and triterpenoid accumulation. Methods: At the end of the growth stage, 0.1 mmol/L and 1.0 mmol/L putrescine (Put), spermine (Spm), and spermidine (Spd) were added to the suspension cells of B. platyphylla, the triterpenoid content and gene expression of lupeol synthase (LUS) related to the synthesis of triterpenoid were analyzed by chemical colorimetry and RT-PCR, respectively. Results: Except for the treatment with 1.0 mmol/L Spm, the cell viability, dry weight, and triterpenoid production in B. platyphylla suspension cells induced by Put and Spd were increased, and the dry weight and triterpenoid production of B. platyphylla cells were increasing with the extension of polyamine treatment time. Among them, dry weight and triterpenoid production were the highest after 2 d Put induction, which were 38.89% and 116.35% higher than those in the control, respectively. Triterpenoid production in B. platyphylla cells induced by polyamine was further confirmed by RT-PCR of LUS gene. Conclusion: Put could effectively enhance the growth of B. platyphylla suspension cells and triterpenoid accumulation.

Interaction between putrescine and hydrogen peroxide on regulating triterpenoid production in Betula platyphylla / 中草药

Objective: To analyze the interaction between putrescine (Put) and hydrogen peroxide (H2O2) on the regulation of triterpenoid production in Betula platyphylla suspension cell culture. Methods: Put (1 mmol/L), H2O2 (0.1 mmol/L), and fungal elicitors (40 μg/mL) were added into the eight-day-old B. platyphylla suspension cell culture, and the changes of triterpenoid content, polyamine content, and H2O2 fluorescence intensity were analyzed by chemical colorimetry, HPLC, and fluorescence microscopy, respectively. Results: Put (1 mmol/L) increased the H2O2 fluorescence intensity and triterpenoid content in B. platyphylla suspension cell culture. H2O2 (0.1 mmol/L) promoted the triterpenoid production, inhibited the production of spermidine (Spd) and spermine (Spm), and had no effectt on Put content. Under the treatment of 1 mmol/L Put and 0.1 mmol/L H2O2, H2O2 fluorescence intensity and polyamine (PAs) content were between its separated treatment, and triterpenoid content was significantly higher than its separated treatment. Comparing with the fungal elicitor, fungal elicitor and H2O2 scavenger catalase (CAT) induced an increase of 0.62% and 16.05% in Put and Spd content, respectively, while no effect on Spm content. Fungal elicitor and PAs synthesis inhibitor D-arginine (D-arg) decreased the H2O2 fluorescence intensity. Under the treatment of fungal elicitor, the fluorescence intensity of CAT and D-arg, H2O2 and the contents of PAs and triterpenoid were lower than those of fungal elicitor and CAT or D-arg, but triterpenoid content was still higher than that of the control. Conclusion: This can be inferred that the interaction between Put and H2O2 regulates the triterpenoid production in birch suspension cell culture.

Accumulative characteristics of triterpenoid precursor and different triterpenoid branch products in Betula platyphylla / 中草药

Objective: Using three-year-old birch (Betula platyphylla) as the experimental material to study the changes of the accumulation of triterpenoid precursor and triterpenoid components in birch, and to analyze the fluctuation characteristics of triterpenoid component accumulation. Methods: Eight compositions, such as the total triterpene, betulin, flavonoids, etc in twig barks and leaves, were analyzed by HPLC and UV spectrophotometer. Results: The accumulative characteristics of each component in triterpenoid substances was significantly different between the twig barks and leaves. During the growing seasons (May-September), the accumulation of 2, 3-oxidized squalene, betulinic acid, betulin, oleanolic acid, dammarenediol, sterol, and total triterpene reached the peak in July-September. Among these, 2, 3-oxidized squalene had a lower content and its highest content was 0.067 mg/g in July. The content of betulinic acid was 8.34 mg/g, while the content of oleanolic acid reached 37.51 mg/g in September. The highest content of dammarenediol was 14.18 mg/g and the total triterpene content was 168.17 mg/g in July. The higher concentration of total triterpene in the leaves was 126.11 mg/g in August, while the contents of 2, 3-oxidized squalene, betulin, and oleanolic acid were very low and not detected. The accumulated peaks of flavonoids in twig barks and leaves were 53.02 and 58.49 mg/g, respectively in May and August. The fluctuation analysis showed that the accumulation of triterpenoid components did not coincide with that of total triterpene in twig barks and leaves,. The total triterpene, dammarenediol, oleanolic acid, and betulinic acid showed the S-shaped curve in twig barks, and the accumulation of betulin showed the inverted "V" curve, the while trend of flavonoid accumulation was contrary to the total triterpene. The trend of the total triterpene accumulation was similar to that of flavonoids, while the trends of dammarenediol and 2, 3-oxidized squalene accumulation were similar. Conclusion: This research identifies the accumulation and distribution characteristics of triterpenoid and flavonoids in the twig barks and leaves of three-year-old birch, and lays the foundation for triterpenoid resources discovering, sustainable use, and metabolic regulation in twig barks and leaves of birch.

A Clinical Study on the Efficacy of Cosmetics Containing the Root of Ephedra sinica and the Bark of Betula platyphylla var. japonica Related to Skin Furrows / 대한피부과학회지

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.

Antitumor activity and immunoregulatory effect of triterpenes isolated from betulaplatyphylla / 中国免疫学杂志

To study the antitumor activity and immunoregulatory effect of Triterpenes isolated from Betula Platyphylla (TBP)Methods: Inhihition rate (IR) on tumor growth was determined by the mice with transplantable tumors (melanoma B16, Sarcoma 180, Lewislung carcinoma and Ehrlich ascites cancer), splenocyte proliferation induced by ConA was measured by H-TdR incorporation; cytotoxicity ofmacrophage was observed by 3H-TdR release; activity of TNF and NK cell was determined by dye (natural red)release. Results: IR of 0.80 g/kg and 1.20 g/kg TBP on all of above tumor strains were greater than30％ in vivo. TBPdoes not show significant effect of splenocyte Prolifera-tion and NK cell activity, but significantly promoted the activity of TNF producted by macroghage and splenocyte. TBP also increase the cyto-toxicity of macrophage. Conclusion:TBP shows potent antitumor effect in vivo. Promoting nonspecific immunoactivity is one of the antitumormechanism of TBP.