Quality Evaluation of Tremella fuciformis Polysaccharides from Different Sources / 中国实验方剂学杂志

Objective:To establish a method for the determination of polysaccharide and monosaccharide composition of Tremella fuciformis, and to analyze the difference of polysaccharide content in T. fuciformis from different sources and cultivation methods, so as to provide reference for the quality determination.Method:High performance size exclusion chromatography coupled with multi-angle laser light scattering and refractive index detection (HPSEC-MALLS-RID) was employed to determine the content and relative molecular weight distribution of T. fuciformis polysaccharides. The monosaccharide types and proportions of T. fuciformis polysaccharides were analyzed by 1-phenyl-3-methyl-5-pyrazolone (PMP) precolumn derivative high performance liquid chromatography (HPLC).Result:The weight-average relative molecular weight (Mw) and the content of polysaccharides in T. fuciformis cultivated by cut-log from different sources were distributed in 2.618×106-3.503×106 Da and 307.12-609.06 g·kg-1, respectively. These two parameters of polysaccharides in T. fuciformis with substitute cultivation from different sources were 2.723×106-3.886×106 Da and 366.38-647.37 g·kg-1, respectively. The T. fuciformis polysaccharides mainly consisted of mannose, glucuronic acid, glucose, galactose, xylose and fucose, their ratios in samples with cut-log and substitute cultivation were 4.4∶0.7∶1.0∶0.2∶1.4∶1.6 and 4.4∶0.8∶1.0∶0.1∶1.5∶1.5, respectively. The contents of the above six monosaccharides in 39 batches of T. fuciformis from different sources were mannose of 36.71-191.31 g·kg-1, glucose of 10.46-76.10 g·kg-1, galactose of 1.00-6.72 g·kg-1, xylose of 16.73-70.54 g·kg-1, glucuronic acid of 9.74-32.12 g·kg-1, fucose of 17.16-68.20 g·kg-1.Conclusion:The content of polysaccharides in T. fuciformis from different sources has a certain difference, the developed method can be used as a routine method for the quality evaluation of polysaccharides in T. fuciformis.

Physicochemical Properties, Microstructure, and Antioxidant and Anti-inflammation Effects of Four Major Polysaccharides from Tremella / 中国药学杂志

OBJECTIVE: To isolate four polysaccharides from Tremella and study their physicochemical properties, microstructures, antioxidant activities and anti-inflammation effects. METHODS: The polysaccharides were purified using water extraction method, DEAE-Sepharose CL-6B and Sephadex G-100. The physicochemical properties and microstructures were studied using high-performance size-exclusion chromatography (HPSEC), gas chromatography (GC), Fourier transform infrared spectroscopy (FTIR) and transmission electron microscopy (TEM) analysis. Anti-inflammation activities of the polysaccharides were identified using real time quantitative PCR. RESULTS: Four kinds of major polysaccharides (TABW-Ⅱ, TFBW-Ⅱ, TEPW-Ⅱ and TSPW-) were purified from water-extraction crude polysaccharides from Tremella. TABW-Ⅱ, TFBW-Ⅱ, TEPW-Ⅱ and TSPW- were homogeneous with relative molecular mass 26×103, 11×103, 21×103 and 356×103, respectively, and they consisted mainly of glucose and mannose. TEM reveale that their molecular morphologies were linear, and TSPW- had more extended structure with entangled chains. Compared with vitamin C (Vc), TSPW- showed more noticeable scavenging effects of superoxide (EC50=0.127 mg•mL-1) and hydroxyl (EC50=0.088 mg•mL-1) radicals (P<0.05). TSPW- also showed the highest inhibitory effects of TNF-α, IL-6 and COX-2 mRNA expressions in LPS-stimulated RAW 264.7 mouse macrophage cells, and the inhibitory effects were more than 95% at 150 μg•mL-1 TSPW-. CONCLUSION: The results indicate that TSPW- with entangled and extended liner structures have more obvious antioxidant and anti-inflammation effects than other major polysaccharides from Tremella family. Furthermore, this study indicates the potential utilization of Tremella polysaccharides in anti-inflammatory agents.

An Enzymolysis-Assisted Agrobacterium tumefaciens-Mediated Transformation Method for the Yeast-Like Cells of Tremella fuciformis

Agrobacterium tumefaciens-mediated transformation (ATMT), as a simple and versatile method, achieves successful transformation in the yeast-like cells (YLCs) of Tremella fuciformis with lower efficiency. Establishment of a more efficient transformation system of YLCs is important for functional genomics research and biotechnological application. In this study, an enzymolysis-assisted ATMT method was developed. The degradation degree of YLCs depends on the concentration and digestion time of Lywallzyme. Lower concentration (≤0.1%) of Lywallzyme was capable of formation of limited wounds on the surface of YLCs and has less influence on their growth. In addition, there is no significant difference of YLCs growth among groups treated with 0.1% Lywallzyme for different time. The binary vector pGEH under the control of T. fuciformis glyceraldehyde-3-phosphate dehydrogenase gene (gpd) promoter was utilized to transform the enzymolytic wounded YLCs with different concentrations and digestion time. The results of PCR, Southern blot, quantitative real-time PCR (qRT-PCR) and fluorescence microscopy revealed that the T-DNA was integrated into the YLCs genome, suggesting an efficient enzymolysis-assisted ATMT method of YLCs was established. The highest transformation frequency reached 1200 transformants per 106 YLCs by 0.05% (w/v) Lywallzyme digestion for 15 min, and the transformants were genetically stable. Compared with the mechanical wounding methods, enzymolytic wounding is thought to be a tender, safer and more effective method.

Chemical constituents from fruiting bodies of Tremella sanguinea / 中国中药杂志

The chemical constituents from the fruiting bodies of Tremella sanguinea were separated and purified by column chromatography on silica gel,ODS,Sephadex LH-20,and RP-HPLC. The structures of the isolated compounds were identified on the basis of physicochemical properties and spectroscopic data analysis,as well as comparisons with the data reported in the literature. Sixteen compounds were isolated from the 90% ethanol extract of the fruiting bodies of T. sanguinea,which were identified as( 22 E)-5α,8α-epidioxy-24-methyl-cholesta-6,9( 11),22-trien-3β-ol( 1),( 22 E)-5α,8α-epidioxyergosta-6,22-dien-3β-ol( 2),cerevisterol( 3),ergosta-7-ene-3β,5α,6β-triol( 4),( 22 E)-6β-methoxyergosta-7,22-diene-3β,5α-diol( 5),ergosta-7-en-3β-ol( 6),4-hydroxy-methylincisterol( 7),2-pyrrolidone( 8),nicotinamide( 9),1-( 3-indolyl)-3-dihydroxypropan-1-one( 10),yangambin( 11),linoleic acid( 12),( 9 Z,12 Z,15 Z)-2,3-dihydroxypropyl octadeca-trienoate( 13),( 9 Z,12 Z)-2,3-dihydroxypropyl-octadeca-dienoate( 14),crypticin B( 15)and 3-phenyllactic acid( 16). All compounds were isolated from T. sanguinea for the first time. Except for compounds 6,9 and 12,the remained compounds were isolated from the genus Tremella for the first time.

Analysis of Curative Effect of LEEP Combined with Tremella Shu in Treating Cervical Precancerous Lesions with HPV Infection / 中国生化药物杂志

Objective To analyze the clinical efficacy of LEEP combined with Tremella Shu in the treatment of cervical cancerous lesions complicated with high-risk human papillomavirus (HPV) infection. Methods From March 2013 to May 2017, 88 patients with cervical precancerous lesions complicated with HPV infection selected from March 2013 to May 2017 in Beichen Hospital of Tianjin were randomly divided into experimental group and control group,44 cases in each group. The control group were treated with LEEP, the experimental group was treated with LEEP combined with Tremella Shu,The HPV viral load, clinical efficacy and quality of life score were compared between the two groups. Results The cure rate was 95.45％ in the experimental group higher than that in the control group(81.81％),the difference was statistically significant (P<0.05). The quality of life score of the experimental group was better than that of the control group,the difference was statistically significant(P<0.05). The HPV viral load in the experimental group was lower than that in the control group, and the difference was statistically significant (P<0.05). Conclusion LEEP treatment combined with Tremella Shu on cervical precancerous lesions with HPV infection of clinical efficacy is significant, can effectively improve the quality of life of patients, reduce HPV viral load, worthy of clinical application.

An Improved Total RNA Extraction Method for White Jelly Mushroom Tremella fuciformis Rich in Polysaccharides

An improved method for extracting high quality and quantity RNA from a jelly mushroom and a dimorphic fungus—Tremella fuciformis which is especially rich in polysaccharides, is described. RNA was extracted from T. fuciformis mycelium M1332 and its parental monokaryotic yeast-like cells Y13 and Y32. The A260/280 and A260/230 ratios were both approximately 2, and the RNA integrity number was larger than 8.9. The yields of RNA were between 108 and 213 µg/g fresh wt. Downstream molecular applications including reverse transcriptional PCR and quantitative real-time PCR were also performed. This protocol is reliable and may be widely applicable for total RNA extraction from other jelly mushrooms or filamentous fungi rich in polysaccharides.

Optimization of Ultrasonic-assisted Extraction Process of Tremella Polysaccharides Based on Box-Benhnken Response Surface Methodology / 中国药师

Objective:To optimize the ultrasonic-assisted extraction technology of polysaccharide from Tremella by Box-Benhnken response surface methodology .Methods:The single-factor extraction tests and response surface analysis were employed to optimize the technology conditions , including ultrasonic power , ultrasonic time and liquid/solid ratio.Results:The experiment results indicated that the optimal extraction conditions were ultrasonic power of 360 W, ultrasonic time of 23 min and liquid/solid ratio of 45 ∶1 ( ml· g-1 ) . Under the conditions , 6 batches of polysaccharide from Tremella were extracted .The average extraction rate of Tremella polysaccharides was 20.4%±1.8%(n=6).Conclusion: The ultrasonic-assisted extraction process of Tremella polysaccharide using Box-Behnken response surface methodology is simple with good predictability .

Tremella Polysaccharides attenuated sepsis through inhibiting abnormal CD4+CD25 high regulatory T cells in mice / 中国免疫学杂志

Objective:To determine the effects of TPS on peripheral blood Tregs in sepsis mouse induced by burn plus P.aeruginosa infection.Methods: The experimental mice were separated into five groups randomly ,including sham burn group ,burn plus P.aeruginosa infection group ,burn plus P.aeruginosa infection with TPS (50,100,200 mg/kg) treatment group.Peripheral blood Tregs were isolated with Magnetic Microbeads and cultured in vitro from the day after burn (PBD0) to 4 days after burn(PBD4).IL-10, IFN-γ,IL-4 levels in Tregs culture supernatants were determined by sandwich enzyme-linked immunosorbent assays ( ELISA ) . Purification of CD4+CD25high Tregs and CD4+T cells in C57BL/6 mice were administrated by magnetic beads sorting .Tregs and CD4+T cells were cultured in vitro after joining TPS to without TPS cells as a control .The phenotypes of Tregs were analyzed by flow cytometry , and cytokines were measured by ELISA .Results:Vis-a-vis the results of the untreated group ,TPS could markedly decrease IL-4 and IL-10 secretion level and significantly increase the secretion of IFN-γ,and the secretion of IL-10 level and concentration of TPS dose effect.Vis-a-vis the results of the untreated group ,in vitro experiment ,without stimulation of TPS ,CD4+T cell proliferation and IFN-γwere significantly reduced ( P<0.05 ) and IL-4 levels increased significantly;CD4+T cell proliferation and IFN-γwere significantly increased and IL-4 levels were significantly reduced in the group of TPS with antibody-1;there was no significant difference in CD 4+T cell proliferation and the levels of IFN-γand IL-4 in the group of TPS with antibody-2.Conclusion:TPS could inhibit the abnormal ac-tivities of CD4+CD25highTregs in burn with P.aeruginosa infection mice,at least in part via inhibiting IL-10 secretion,and trigger a shift of Th2 to Th1 with activation of CD4+T cells in burn with P.aeruginosa infection mice.

Protection of phosphate esterification polysaccharides from Tremella fuciformis on hematopoietic function in radiation-injured mice / 中草药

Objective: To study the effect of phosphate esterification polysaccharides from Tremella fuciformis (PEPTF) on hematopoietic function in radiation-injured mice. Methods: Rats were randomly divided into six groups: control, model, polysaccharides from T. fuciformis (10 mg/kg) positive control, low-, mid-, and high-PEPTF (10, 30, and 60 mg/kg) groups. Before irradiated with 137Cs γ-ray, rats were ip administered once daily for 3 d. On the day 7 after irradiation, the rats were sacrificed. Colony-forming unit of spleen, number of nucleated cells in bone marrow, number of white blood cells, spleen index, and thymus index were used as observation indexes to investigate the effect of PEPTF on hematopoietic function of mice. Results: Compared with the model group, the number of the nucleated cells in bone marrow, the number of white blood cells, spleen index, and thymus index of mice increased markedly. Conclusion: PEPTF have the protective effect on the hematopoietic function of radiation-injured mice.

Análise preliminar do tratamento de Diabetes mellitus com Polissacarídeo de Tremella fuciformis / Preliminary analysis of the treatment of Diabetes mellitus with Tremella fuciformis polysaccharide

Introdução: O Diabetes mellitus é uma desordem metabólica que ocorre no mundo inteiro, caracterizada por hiperglicemia associada a problemas com secreção ou ação da insulina ou, ainda por alterações no metabolismo intermediário de carboidratos, proteína e lipídeos. O cogumelo Tremella fuciformis (Basideomycota) foi isolado no Brasil e levado para a China onde tem sido produzido e utilizado como alimento por apresentar diversas atividades biológicas como anti-inflamatório, anti-mutagênico, reduz hipertensão, reduz diabetes entre outras propriedades relacionadas com imunidade. A Embrapa de Brasília tornou isolar o cogumelo e enviou a universidade para os testes preliminares. Objetivo: O presente trabalho teve como objetivo induzir a diabetes em ratos e realizar tratamento com polissacarídeo (PS) extraído do cogumelo gelatinoso Tremella fuciformis, desenvolvido em frascos no laboratório. Método: Ratos, no total de 28 animais foram divididos em 4 grupos sendo: 1-Normal) grupo recebendo dieta normal e água; 2-Sacarose) grupo recebendo ração normal e uma solução de sacarose 20% por 65 dias; 3-Sacarose + PS) grupo recebendo ração normal + solução de sacarose (65dias) e por gavagem PS na concentração de 1mmol (nos últimos 20 dias sendo uma vez/semana) ; 4-idem ao grupo 3, entretanto PS com concentração de 2mmol. No período, os ratos (uma vez/semana) foram pesados e submetidos a teste de glicose (fita Accu Check) e, depois sacrificados com dose letal de analgésico e coleta do sangue para testes de glicose, colesterol, triglicérides, creatinina e HDL. Resultados: Os resultados parciais demonstraram que os animais submetidos à dieta com sacarose aumentaram o peso e glicemia. Já os animais submetidos a tratamento com o PS permaneceram com o peso praticamente constante e, redução da glicemia. A creatinina tem sido um parâmetro para diagnosticar problemas renais, mas os animais em todos os tratamentos, não apresentaram diferença estatística. Para colesterol os animais submetidos ao tratamento com o PS apresentaram valores abaixo dos encontrados no grupo com sacarose, entretanto, com triglicérides e HDL apresentaram diminuição, mas houve, aumento em relação aos animais do grupo normal, indicando controle nos lipídeos. Por conclusão os resultados preliminares demonstraram que o PS extraído do cogumelo Tremella fuciformis apresenta grande potencial como produto natural contra diabetes.

Quality control of Tremella fuciformis polysaccharide by HPGPC / 中草药

Objective: To develop a high performance gel permeation chromatography (HPGPC) method for quality control of Tremella fuciformis polysaccharide. Methods: The separations were carried out with two series-wound columns including Shodex Sugar KS-804 (300 mm x 8 mm, 7 μm) and KS-805 (300 mm x 8 mm, 17 μm), water as mobile phase at flow rate 0.8 mL/min, column temperature at 40°C, injection volume 40 μL, and differential index detector. Results: T. fuciformis polysaccharide peak was located within 14.40-15.30 min and corresponding molecular weight was 1.26 × 106-1.39 × 106. The linearity was good during 0.01-2.0 mg/mL (r = 0.999). The polysaccharide concentration was 1.1-1.5 mg/mL and 0.8-1.1 mg/mL with precipitation of T. fuciformis polysaccharide and blue dextran as reference, respectively. LOD and LOQ were 0.8 and 2 μg, respectively. Conclusion: The method is simple, quick, and accurate, and has good repeatability and high sensitivity, which is fitful for quality control.

Anti-apoptosis effect of tremella polysaccharides on cardiomyocytes / 第二军医大学学报

Objective: To observe the anti-apoptosis effect of tremella polysaccharides (TP) on cardiomyocytes by in vitro and in vivo experiments. Methods: TP was extracted by dehydrated alcohol and the TP purity was measured by gas chromatography. The cultured cardiomyocytes of the neonatal rats were randomly assigned to normal group (Group A), apoptosis-induced group (Group B) and TP-pretreatment group (Group T). The morphology and uptake rate of trypan blue were studied after 72 h. The apoptosis index of cardiomyocyte was measured by flow cytometry. In the in vivo study, 100 ICR mice were randomly assigned to 5 groups after paired by body weight as following: Group N (negative control group, peritoneally injected by saline daily); Group C (positive control group, peritoneally injected by D-galactose and saline daily); Group L, M, and H (peritoneally injected by D-galactose and intragastric administered with TP at 100, 200, 400 mg/day, respectively). Mice were sacrificed for histology and biochemistry studies after 8 weeks. Results: Data of cultured cardiomyocytes were as follows: Uptake rate of trypan blue: B>C>A (PC> A (P<0.01). In the animal studies there were no obvious difference in myocardium pathology between groups. The order of the apoptosis indices of myocardium was H

Effect of tremella polysaccharides on the antioxidant ability of experimental aging mice / 中华临床营养杂志

ificantly lower than in both middle and low group(P＜0.05).Conclusion TP may improve the antioxidant ability of experimental aging mice.

Composition and Partial Structure Characterization of Tremella Polysaccharides

Heteropolysaccharides isolated from liquid cultures of nine Tremella species contained 0.3 to 1.2% protein, 2.7 to 5% ash, 0.9 to 3.4% acetyl groups, 76.5 to 84.2% carbohydrates and trace amounts of starch. The polysaccharides in aqueous solution were slightly acidic (pH 5.1 to 5.6). They consisted of the following monomeric sugars: fucose, ribose, xylose, arabinose, mannose, galactose, glucose and glucuronic acid. The backbones of the polysaccharide structures consisted of alpha-(1-->3)-links while the side chains were beta-linked.

Effect of Tremella Polysaccharides on cardiomyocytes in vitro and in vivo / 中华临床营养杂志

@#myocytes.In aging mice induced by D-galactose experiment.TP has anti-apoptosis and anti-oxidation effect in a dose-dependant manner.

The Neuroprotective and Neurotrophic Effects of Tremella fuciformis in PC12h Cells

We examined the neuroprotective and neurotrophic effects of Tremella fuciformis. The neurotrophic effects of the hot water extract of T. fuciformis was evaluated by microscopically monitoring its potency to induce neurite outgrowth in PC12h cells. The hot water extract of T. fuciformis promoted neurite outgrowth in PC12h cells in this study, superior to other natural substances which was reported previously. When cells were treated with the hot water extract of T. fuciformis prior to beta-amyloid peptide treatment (active domain of A peptide 25~35 treated), toxicity was significantly diminished (p<0.01). These results suggest that T. fuciformis might potentially be used as a precautionary agent in neurodegenerative disease, such as Alzheimer's disease, etc.

Antitussive and Antiasthmatic Effect of Compound Liquor from Tremella Aurantialba / 医学研究杂志

Objective To study the antitussive and antiasthmatic effect of compound liquor from Tremella aurantialba. Methods The mice cough model induced by ammonia, guinea pig cough model induced by acitric acid,guinea pig asthma model induced by histamine and acetylcholine, guinea pig asthma model induced by ovalbumin were used in this experiment. Results The compound liquor from Tremella aurantialba could significantly prolong the tussive delitescence,decreas the tussive times of mice and guinea pigs;could significantly prolong the asthmatic delitescence. Conclusions The antitussive and antiasthmatic effects of compound liquor from Tremella aurantialba was obvious.

Preliminary Study on the Effective Anticoagulant Components of the Fermented Products of Mycelium from Tremella aurantialba / 中国中医药信息杂志

Objective To study the effective anticoagulant components of the fermented products of mycelium from Tremella aurantialba 9800. Method To adopt system menstruum deal with fermented products of mycelium from Tremella aurantialba,then mycelium from tremella aurantialba was studied with the clotting time of the thrombogen,the whole blood clotting time in mice and thrombosis experiment in mice. Result Butanol extract of fermented products of mycelium from Tremella aurantialba behave anticoagulant function. Ethyl acetate extract and water extract also behave feeble activity. Conclusion The effective anticoagulant components of the fermented products of mycelium from Tremella aurantialb is butanol extract.

Protective effect of tremella polysaccharides on experimental aging mouse model / 第二军医大学学报

Objective:To observe the protective effect of tremella polysaccharides (TP) on mouse aging model induced by D-galactose. Methods: Forty male ICR mice were divided into 5 groups (Group A: treated with 800 mg/kg TP; Group B: treated by 400 mg/kg TP; Group C: treated by 200 mg/kg TP; Group D: aging control; Group E: normal control). Except for Group E,other groups were injected with D-galactose [120 mg/(kg?d)] for 60 d to establish subacute aging model. Group A,B and C were treated with 800,400 and 200 mg/kg TP respectively via intragastric (i.g.) administration. Sixty days later,the spleen,heart, kidney and brain were collected to determine MDA, LP, SOD, GSH-Px and contents of hydroxyproline by colorimetry. Expression of P21 in the kidney were detected by immunohischemical technique. Proliferation and transformation of spleen lymphocytes induced by ConA were determined by MTT method. Results: Antioxidants SOD and GSH-Px activity of TP treated groups were higher than aging control group (P

The study on the experiment of hepatocarcinoma with homogenous spleenocytes activated by IL-2 and Tremella Polysaccharide / 中国免疫学杂志

Objective:To study the cytotoxicity to tumor cells P815,H22 and B16 in vitro and the treatment effect of mice spleenocytes activated by lymphokine IL-2 and Tremella Polysaccharide,and their effects on main viscerals.Methods:The cytotoxicity of spleenocytes activated by lymphokine IL-2 or IL-2 and Tremella Polysaccharide to P815,H22 and B16 was detected by ?-scintillation counter after 2 hours.Subcutaneous injection of the above-mentioned spleenocytes to the local area of jepatocarcinoma of mice,one injection every two days,5 times totally.To examine tumor weight,size,histological changes and morphological changes in main viscerals.Results:The significantly cytotoxicity to tumor cells of spleenocytes activated by IL-2 and Tremella Polysaccharide was observed.To B16,the cytotoxicity of activated spleenocytes was the strongest.To H22,the cytotoxicity of activated spleenocytes was stronger and to P815,the cytotoxicity was the weakest,the cytotoxicity of TP-LAK cells was significantly stronger than that of LAK cells( P