Preparation of Diosgenin from Dioscorea zingiberensis： A Review / 中国实验方剂学杂志

Direct acid hydrolysis of Dioscorea zingiberensis rhizomes for preparation of diosgenin is wildly used in the traditional industry， which uses a large amount of inorganic acid catalysts， with high wastewater discharge and serious environmental pollution. Therefore， exploring clean and efficient preparation methods and processes has become an inevitable choice to realize the sustainable development of industrial production of diosgenin. Herein， the author reviewed and analyzed the research progress and problems of enzymatic hydrolysis， microbial transformation and modified acid hydrolysis in the preparation of diosgenin from D. zingiberensis rhizomes during the last ten years， and their application prospects are analyzed. Enzymatic hydrolysis has mild reaction conditions， but the yield of diosgenin is low， the economic cost is high， and the purification process of active enzyme is complicated. Microorganism shows specific activity to the substrate and high efficiency for diosgenin production， and microbial transformation is clean and environmentally friendly， but microbial transformation is time-consuming and the metabolic intermediates are complicated. For the modified acid hydrolysis， two-phase acid hydrolysis can reduce the amount of acid catalyst， and sulfonic acid-functionalized ionic liquid displays good recyclable performance by replacing the traditional inorganic acid， however， the wastewater discharge should still be considered. Solid acid catalysts are non-corrosive and easy to be recycled， but the need to use ethanol as the reaction solvent has certain safety hazards， and the catalyst preparation process is cumbersome. In conclusion， exploring clean and efficient conversion methods is an important research trend for preparation of diosgenin from D. zingiberensis rhizomes. For the enzymatic hydrolysis， the key glycoside hydrolases in the bioconversion process should be explored in depth， the conversion pathway of enzymatic saponins and enzyme specificity should be fully elucidated， and efforts should be made to improve the efficiency of enzymatic hydrolysis. For the microbial transformation， we should accelerate its industrial application process based on selecting and breeding efficient transformation strains， and optimizing stable transformation systems and processes， and in-depth investigation of the mechanism of microbial transformation， fully elucidating the specific key hydrolases and its catalytic properties， and striving to improve the efficiency of microbial transformation. For the modified acid hydrolysis， novel acid catalytic system with simple structure， stable performance and good biodegradability should be explored and applied， which can effectively solve the problems of environmental pollution and production safety.

Effects of Total Saponins from Dioscorea Nipponica Makino on Monosodium Urate-Induced M1-Polarized Macrophages through Arachidonic Acid Signaling Pathway: An in vitro Study / 中国结合医学杂志

OBJECTIVE@#To investigate and reveal the underlying mechanism of the effect of total saponins from Dioscoreae nipponica Makino (TSDN) on the arachidonic acid pathway in monosodium urate (MSU) crystal-induced M1-polarized macrophages.@*METHODS@#M1 polarization of RAW264.7 cells were induced by 1 µ g/mL lipopolysaccharide (LPS). The methylthiazolyldiphenyl-tetrazolium bromide method was then used to screen the concentration of TSDN. MSU (500 µ g/mL) was used to induce the gouty arthritis model. Afterwards, 10 µ g/L TSDN and 8 µ mol/L celecoxib, which was used as a positive control, were added to the above LPS and MSU-induced cells for 24 h. The mRNA and protein expressions of cyclooxygenase (COX) 2, 5-lipoxygenase (5-LOX), microsomal prostaglandin E synthase derived eicosanoids (mPGES)-1, leukotriene B (LTB)4, cytochrome P450 (CYP) 4A, and prostaglandin E2 (PGE2) were tested by real-time polymerase chain reaction and Western blotting, respectively. The enzyme-linked immunosorbent assay was used to test the contents of M1 markers, including inducible nitric oxid synthase (NOS) 2, CD80, and CD86.@*RESULTS@#TSDN inhibited the proliferation of M1 macrophages and decreased both the mRNA and protein expressions of COX2, 5-LOX, CYP4A, LTB4, and PGE2 (P<0.01) while increased the mRNA and protein expression of mPGES-1 (P<0.05 or P<0.01). TSDN could also significantly decrease the contents of NOS2, CD80, and CD86 (P<0.01).@*CONCLUSION@#TSDN has an anti-inflammation effect on gouty arthritis in an in vitro model by regulating arachidonic acid signaling pathway.

GC-MS Metabolites and Gene Expression Characteristics in Fibrous Roots of Dioscorea zingiberensis in Response to Low Phosphorus Stress / 中国实验方剂学杂志

ObjectiveTo investigate the metabolites and gene expression characteristics in fibrous roots of Dioscorea zingiberensis in response to low phosphorus stress. MethodThe severe stress group， the moderate stress group， and the normal group were set up to stimulate the low phosphorus stress experiment. The fibrous roots of D. zingiberensis were collected during initial stress. The metabolites and transcriptomic characteristics were analyzed by gas chromatography-mass spectrometry （GC-MS） derivatization and RNA-seq techniques. Through multivariate statistical analysis of metabolites treated by different methods，functional analysis of differentially expressed genes， and data mining， the metabolism markers produced in fibrous roots of D. zingiberensis under low phosphorus stress were screened out， and the metabolic pathway characteristics of different genes were analyzed. ResultA total of 116 GC-MS metabolites were detected from the fibrous roots of D. zingiberensis. The metabolic characteristics of fibrous roots of D. zingiberensis under different low phosphorus treatments were obviously different. Orthogonal partial least squares discriminant analysis（OPLS-DA） model was used to screen six differential metabolites represented by sugars and alcohols from metabolites of fibrous roots treated with different methods，and these components were presumedly metabolism markers of fibrous roots of D. zingiberensis in response to low phosphorus stress. The differential genes screened out from the severe stress group and the normal group were mainly enriched in peroxidase pathway，phosphate and hypophosphate metabolism pathway，while the differential genes screened out from the severe stress group and the moderate stress group were mainly enriched in glutathione metabolism pathway and phosphopentose pathway. A total of 177 differential genes in response to low phosphorus stress were screened out from fibrous roots， involving many pathways such as terpenoid skeleton and inositol biosynthesis，which was consistent with the fact that the metabolic differential components in fibrous roots in response to low phosphorus stress were mainly saccharides and inositol. ConclusionThe metabolites and gene expression in fibrous roots of D. zingiberensis responded to low phosphorus stress，and the differential metabolites were closely related to differentially expressed genes. This study is expected to provide a theoretical basis for the research on the molecular mechanism of D. zingiberensis in response to low phosphorus stress.

Lowland and highland Varieties of Dioscorea esculenta tubers stimulate growth of Lactobacillus sp. over enterotoxigenic E. coli in vitro

@#<p style="text-align: justify;"><strong>Objectives:</strong> Probiotic supplementation often only leads to transient improvement in the gut microbiome. Potential prebiotics, such as the oligosaccharide-rich varieties of Dioscorea esculenta tubers, can potentially bridge the gap between supplementation and persistent colonization. Thus, this study aimed to assess the ability of D. esculenta tubers to promote the growth of probiotic Lactobacillus sp. in vitro selectively.</p><p style="text-align: justify;"><strong>Methods:</strong> The prebiotic activity of the selected varieties of Dioscorea esculenta tubers was evaluated via compe titive growth assay, wherein the ratios of probiotic Lactobacillus sp. over enterotoxigenic Escherichia coli (ETEC) or "prebiotic ratios" were compared following treatment.</p><p style="text-align: justify;"><strong>Results:</strong> Negative control (0.9% NaCl solution) produced a ratio of 0.88, Lowland and Highland varieties produced ratios of 1.26 and 1.29, respectively, and positive control (inulin) produced 1.54. The two varieties had comparable ratios to one another (p > 0.05), and significantly higher ratios than the negative control (p < 0.05). Both varieties have significant prebiotic activity. Compared to inulin, the two varieties' prebiotic activity was 84% as effective.</p><p style="text-align: justify;"><strong>Conclusion:</strong> Overall, the tubers promoted the growth of Lactobacillus sp. over ETEC. The crude tuber samples, given their availability and affordability, can be easily integrated into the local diet to contribute to the improvement of the general population's health.</p>

Experimental Study on Enhancing the Antitussive Efficacy and Reducing the Toxicity of Dioscorea bulbifera L. by the Processing Technology / 中国药学杂志

OBJECTIVE: To investigate the effects of different processing methods on antitussive and hepatotoxicity of Dioscorea bulbifera L.. METHODS The raw and processed products of Dioscorea bulbifera L., fried Dioscorea bulbifera L. (FD), wine-fried Dioscorea bulbifera L. (WD), liquorice liquid-fried Dioscorea bulbifera L. (LD) and angelicae liquid-fried Dioscorea bulbifera L. (AD) were administered to mice at a dose of 1.7 g•kg-1 for 7 d. The effects of processing on the cough relief of Dioscorea bulbifera L. were evaluated by the cough test induced by concentrated ammonia water. The detoxification effect and the preliminary mechanism of processing on Dioscorea bulbifera L. were evaluated by the detection and analysis of the relevant indicators. RESULTS: Compared with the RD group, LD and AD group had longer cough latency (P0.05). Compared with the AD group, there were more coughing times in LD group (P0.05) in mice induced by concentrated ammonia water. The intervention of WD only had significant reversal effect on the low GST level of liver (P0.05). CONCLUSION: Processing with Angelica sinensis and licorice could enhance the antitussive effect of Dioscorea bulbifera L. and reduce its hepatotoxicity. The mechanism of attenuation may be related to the inhibition of lipid peroxidation and the enhancement of antioxidant level in liver. The antitussive effect of two kinds of processing, stir-fried and wine-fried, is only effective, but no synergistic effect, no toxic effect and no attenuation effect are found.

Chemical constituents and antitumor activity from stems and leaves of Dioscorea opposita / 中草药

Objective: To investigate the chemical constituents from the stems and leaves of Dioscorea opposita. Methods: The compounds were isolated and purified by various column chromatographies, and their structures were identified by physiochemical properties and spectroscopic data. The effects of the compounds on the proliferation of human breast cancer MCF-7 cells and human liver cancer HepG2 cells were investigated. Results: Twenty compounds were isolated from the 50% acetone extract of the stems and leaves of D. opposita, which were identified as 1H-indazole (1), 1H-indole-3-carboxaldehyde (2), 1H-indole-3- carboxylic acid (3), 3-(2-oxopropyl)-3-hydroxy-indolin-2-one (4), thymidine (5), uridine (6), 3-hydroxy-3-(2-oxopropyl)-2,4-(1H,3H)- quinolinedione (7), hematinic acid (8), allantoin (9), 2-ethyl-3-methyl-maleimide-N-β-D-glucopyranoside (10), paulownin (11), (+)-8-hydroxypinoresinol (12), (+)-syringaresinol (13), (-)-dihydrodehydrodiconiferyl alcohol (14), (7R,8S)-dihydrodehydro- diconiferyl alcohol-4-O-β-D-glucopyranoside (15), (2E,6S)-6,7-dihydroxy-3,7-dimethyl-2-octenoic acid (16), (2E,4S)-4-hydroxy-2- nonenoic acid (17), (2E,6S)-6-hydroxy-2,6-dimethyl-2,7-dienoic acid (18), amarantholidoside IV (19), (9Z,11E)-13-methoxy- 9,11-octadecadienoic acid methyl ester (20), and their effects on proliferation of MCF-7 cells and HepG2 cells were investigated. compounds 1, 4, 8, 11-15, 18, 20 at a dose of 25μmol/L inhibited the proliferation of MCF-7 cells, and compounds 1, 4, 8, 12-14, 18, 20 at a dose of 25 μmol/L inhibited the proliferation of HepG2 cells. Conclusion: Compounds 1, 3-5, 10-12 and 15-20 are isolated from this plant for the first time, compounds 1, 4,8, 11-15, 18, 20 inhibited the the proliferation of MCF-7 and HepG2 cellssignificantly at certain concentration, showing protent antitumor activity.

Phytoglycoprotein isolated from Dioscorea batatas Decne promotes intestinal epithelial wound healing / 中国天然药物

Dioscorea batatas Decne (DBD) has been used to heal various illnesses of the kidney and intestine as an herbal medicine in Asia. As a source of therapeutic agents, many glycoproteins have been isolated from mushrooms and plants, but the functional role of glycoprotein in intestinal epithelial wound healing has not been understood yet. In the present study, we investigated the wound healing potentials of the 30 kDa glycoprotein (DBD glycoprotein) isolated from DBD in human intestinal epithelial (INT-407) cells. We found that DBD glycoprotein (100 μg·mL) significantly increased the motility of INT-407 cells for 24 h by activating protein kinase C (PKC). DBD glycoprotein stimulated the activation of p38 mitogen-activated protein kinase (MAPK), which is responsible for the phosphorylation of NF-κB inhibitor α (IκBα). DBD glycoprotein increased the level of profilin-1 (PFN1), α-actinin and F-actin expression via activation of transcription factor, nuclear factor-kappa B (NF-κB) during its promotion of cell migration. Experimental mouse colitis was induced by adding dextran sulfate sodium (DSS) to the drinking water at a concentration of 4% (W/V) for 7 days. We figured out that administration of DBD glycoprotein (10 and 20 mg·kg) lowers the levels of disease activity index and histological inflammation in DSS-treated ICR mice. In this regard, we suggest that DBD glycoprotein has ability to promote the F-actin-related migration signaling events via activation of PKC and NF-κB in intestinal epithelial cells and prevent inflammatory bowel disease.

The crucial role of metabolic regulation in differential hepatotoxicity induced by furanoids in Dioscorea bulbifera / 中国天然药物

Diterpenoid lactones (DLs), a group of furan-containing compounds found in Dioscorea bulbifera L. (DB), have been reported to be associated with hepatotoxicity. Different hepatotoxicities of these DLs have been observed in vitro, but reasonable explanations for the differential hepatotoxicity have not been provided. Herein, the present study aimed to confirm the potential factors that contribute to varied hepatotoxicity of four representative DLs (diosbulbins A, B, C, F). In vitro toxic effects were evaluated in various cell models and the interactions between DLs and CYP3A4 at the atomic level were simulated by molecular docking. Results showed that DLs exhibited varied cytotoxicities, and that CYP3A4 played a modulatory role in this process. Moreover, structural variation may cause different affinities between DLs and CYP3A4, which was positively correlated with the observation of cytotoxicity. In addition, analysis of the glutathione (GSH) conjugates indicated that reactive intermediates were formed by metabolic oxidation that occurred on the furan moiety of DLs, whereas, GSH consumption analysis reflected the consistency between the reactive metabolites and the hepatotoxicity. Collectively, our findings illustrated that the metabolic regulation played a crucial role in generating the varied hepatotoxicity of DLs.

Anti-inflammatory and analgesic effects of total saponin of Dioscorea / 中国临床药理学与治疗学

AIM: To study the anti-inflammatory and analgesic effects of total saponin of Dioscorea(TSD). METHODS: The rat acute gouty arthritis (AGA) and the mice air-pouch inflammation models were induced by monosodium urate(MSU), the pain models were adopted including the hot-plate and acetic acid-induced writhing test in mice. Then the content of IL-1β, IL-18 and PGE2 levels were determined by ELISA. RESULTS:TSD significantly inhibited the MSU-induced ankle swelling and air pouch inflammation in mice(P<0.01 or P<0.05). Meanwhile, TSD markedly reduced the levels of IL-1β and IL-18 in synovial fluid exudates and IL-1β and PGE2 in air pouch exudates (P<0.01 or P<0.05).In addition, TSD remarkably suppressed acetic acid-induced writhing response and prolonged pain threshold in hot plate assay(P<0.01 or P<0.05).CONCLUSION: TSD has anti-inflammatory and analgesic effects, which are mainly relevant to the inhibition of the synthesis and release of inflammatory factors IL-1β, IL-18 and PGE2.

Viabiliadade tecnologica da farinha de inhame na formulação de biscoito sem glúten / Technical feedback of inhame flour in the formulation of gluten free biscuit

O inhame apresenta características funcionais e tecnológicas que o tornam útil na elaboração de produtos para grupos específicos, como os portadores de doença celíaca. O objetivo do presente foi estudar a farinha do inhame para a formulação de biscoito funcional isento de glúten. O mesmo foi elaborado a partir de uma receita sem glúten, com a utilização da farinha de inhame. Foi elaborada a ficha técnica e tabela nutricional. A análise microbiológica mostrou que os biscoitos estavam dentro dos padrões legais. A análise sensorial foi testada e aprovada nos quesitos de cor, aparência, textura e odor. A intenção de compra foi de 54%, devido ao sabor residual amargo. O resultado da avaliação global foi de 84%. O biscoito mostrou-se promissor, devendo ser trabalhado o aspecto sabor a fim de melhorar sua aceitabilidade e intenção de compra.

Cloning and Bioinformatics Analysis on CDS of SUS Gene in Dioscorea opposita / 中国实验方剂学杂志

Objective: To clone CDS sequence of Dioscorea opposita SUS gene and analyze the protein structure,in order to provide a theoretical basis for the regulation mechanism of SUS gene and the synthesis mechanism of D.opposita polysaccharides.Method: Total RNA in D.opposita was extracted and reverse-transcribed into first strand of cDNA.Specific primers were designed according to an annotated SUS gene sequence obtained from the laboratory D.opposita genome database,and the coding region of the SUS gene was obtained by gene cloning technique and the protein sequence characteristics were analyzed by protein prediction analysis software.Result: A 2 448 bp gene sequence was cloned with a complete open reading frame (ORF).The gene was named DoSUS1.The formula of protein encoded by DoSUS1 gene in D.opposita was C4209H6534N1115O1205S23,and the molecular weight was 9 788.32,the total number of amino acids was 815,the theory isoelectric point (PI) was 6.10,the extinction coefficient was 110 505,the aliphatic index (AI) was 94.15,the instability index was 32.18,and the grand average of hydropathicity (GRAVY) was-0.225.It was a stable and soluble acidic protein.There were phosphorylation sites in the DoSUS1 amino acid sequence,with no transmembrane region and signal peptide.The secondary structure and the tertiary structure showed that DoSUS1 was an α class protein.Functional domain predictions showed that DoSUS1 had sucrose synthesis domain and glycosyl transfer domain.The homology alignment showed that the amino acid sequence of DoSUS1 was more than 80% similar to the amino acid sequence of the aligned monocots.The phylogenetic tree showed that DoSUS1 was closely related to SUS of monocotyledon evolution.Conclusion: The coding sequence of SUS gene was cloned from D.opposita for the first time,and its protein structure was analyzed to lay a foundation for further studying the roles of SUS in the growth and polysaccharide synthesis of D.opposita.

Comparison of Diosgenin Content in Wild Dioscorea zingiberensis from Different Habitats / 中国药房

OBJECTIVE：To establish a method for the content determination of diosgenin in Dioscorea zingiberensis，and to compare the contents of diosgenin in wild D. zingiberensis from different habitats. METHODS：HPLC method was adopted. The determination was performed on Inertsil ODS-3 column with mobile phase consisted of acetonitrile-water （50 ∶ 50，V/V） at the flow rate of 1.0 mL/min. The column temperature was set at 30 ℃. The detection wavelength was set at 203 nm，and sample size was 10 μL. Established method was used to determine the contents of diosgenin in wild D. zingiberensis from different habitats （Shaanxi Shangluo，Shaanxi Ankang，Henan Neixiang，Yunnan Xuanwei，Sichuan Deyang，Hubei Shiyan，Hunan Zhangjiajie，Hunan Changde）. The differences of diosgenin content were compared. RESULTS：The linear range of diosgenin were 4.16-165.6 μg/mL （r＝     0.999 9）. RSDs of precision，reproducible and stability tests were all lower than 2％ （n＝5 or n＝6）. The average recovery of diosgenin was 101.18％ （RSD＝1.27％，n＝6）. The content of diosgenin in wild D. zingiberensis from different habitats were in descending order，i.e. Sichuan Deyang （1 405.36 μg/g）＞Shaanxi Shangluo （1 201.79 μg/g）＞Hunan Zhangjiajie （1 035.18 μg/g）＞Shaanxi Ankang （632.64 μg/g）＞Hunan Changde （598.64 μg/g）＞Yunnan Xuanwei （425.34 μg/g）＞Henan Neixiang （350.13         μg/g）＞Hubei Shiyan （338.39 μg/g）. CONCLUSIONS：Established method is simple，accurate and suitable for the content determination of diosgenin in D. zingiberensis. The contents of diosgenin in wild D. zingiberensis from different habitats are different significantly，and the content of diosgenin in the samples from Sichuan Deyang is the highest.

Dioscorea japonica Thunb. Ethanolic Extract Attenuated Oxazolone-Induced Atopic Dermatitis-like Skin Lesions in BALB/c Mice

The rhizomes of Dioscorea japonica Thunb. are widely consumed as food and also used to treat diabetes and polyuria in Korea. This study was undertaken to study the anti-atopic dermatitis effects of a 95% ethanolic extract (DJE) of D. japonica in an oxazolone-stimulated murine model of atopic dermatitis (AD). The therapeutic effects of DJE on AD-like skin lesions were assessed on both ears. DJE (1%) or dexamethasone (0.5%; the positive control) were applied to skin lesions for three weeks. Serum levels of IgE and IL-4 were assessed by ELISA (enzyme-linked immunosorbent assay). Histopathological examinations were performed by hematoxylin and eosin (H&E) and toluidine blue staining and revealed DJE significantly reduced dermal thickness and inflammatory cell infiltration when applied to oxazolone-treated ear skin. DJE-treated AD mice also showed lower serum levels of IgE and IL-4 than oxazolone-stimulated controls. Our findings demonstrate DJE might be a useful safe, topical agent for the treatment of atopic diseases.

Anti-inflammatory Effect of Total Saponin Fraction from Dioscorea nipponica Makino on Gouty Arthritis and Its Influence on NALP3 Inflammasome / 中国结合医学杂志

OBJECTIVE@#To investigate the mechanism of Chinese herbal medicine Dioscorea nipponica for the treatment of monosodium urate crystals-induced gouty arthritis (GA) in rats.@*METHODS@#Sixty male Wistar rats were divided into 6 groups: normal, model, indomethacin and three total saponin (900, 300 and 100 mg/kg) groups. The liver, kidney and serum levels of lysosomal enzymes, antioxidant capacities, and inflammatory factors were measured. In addition, the mRNA and protein levels of the NALP3 inflammasome components in the mononuclear cells of rats' peripheral blood were analyzed using real-time polymerase chain reaction and Western blotting methods, respectively.@*RESULTS@#Total saponins groups could reduce the activities of β-galactosidase, β-N acetyl glucosamine enzyme, β-glucuronidase, acid phosphatase, and malonaldehyde as well as the contents of TNF-α, IL-1β and IL-8 (all P<0.05). They could also increase the activities of glutathione peroxidase and total superoxide dismutase (both P<0.05). Further studies showed that total saponins groups of high, middle and low doses could all increase the mRNA and protein levels of caspase-1, adapter apoptosis-associated speck-like protein containing a caspase-recruitment domain (ASC) and NALP3 in the mononuclear cells of peripheral blood (all P<0.05).@*CONCLUSION@#Dioscorea nipponica may treat GA by regulating lysosomal enzymes, antioxidant capacities and the NALP3 inflammasome.

Symptomatic Hypocalcemia Associated with Dioscorea tokoro Toxicity

Dioscorea tokoro has long been used in Korean traditional medicine as a pain killer and anti-inflammatory agent. A 53-year-old male who consumed water that had been boiled with raw tubers of D. tokoro as tea presented with numbness and spasm of both hands and feet. Laboratory results showed hypocalcemia, hypoparathyroidism, and vitamin D insufficiency. During his hospital stay, colitis, acute kidney injury, and toxic encephalopathy developed. The patient received calcium gluconate intravenous infusion and oral calcium carbonate with alfacalcidol. His symptoms improved gradually, but hypocalcemia persisted despite the calcium supplementation. We suggest that ingestion of inappropriately prepared D. tokoro can cause symptomatic hypocalcemia in patients with unbalanced calcium homeostasis.

Clinical analysis of 45 cases of drug-induced liver injury induced by bulbil of dioscorea bulbifera L / 国际中医中药杂志

Objective To explore the characteristics, diagnosis, treatment and prognosis of liver injury caused by the deficiency of dioscorea bulbifera L.. Methods The general data, clinical manifestation and laboratory examination of 45 cases of liver injury diagnosed as Yoshimoto associated liver injury from November 2014 to June 2017 were classified and reviewed with the standards of drug liver injury classification recommended by the Council of international medical organizations. Results The number of male patients was 26, and female 19. The medication time ranged from 1 week to 2 years and the main biochemical performance was abnormal, namely ALT, AST, TBil, DBil, ALP and GGT. Of the 45 cases, the average values of ALT, AST were 608.11 ± 411.30 U/L and 505.38 ± 342.15 U/L. The TBil of 42 case rised with the mean value 170.10 ± 136.86 μmol/L, and the ALP of 22 cases with 182.38 ± 55.15 U/L. The GGT of 43 cases rised with the mean 223.12 ± 131.85 U/L. Clinical classification included 38 cases were liver cell injury, none was cholestasis, 5 mixed types and 2 cases of liver biochemical examination abnormality. One patient died while the other patients recovered. Conclusions Although the pathogenesis of the liver cell induced injury type with dioscorea bulbifera L. remains unclear, the reasonable and appropriate use of medication and regular liver biochemical tests is necessary.

Enhancement Study of Macrophage Phagocytic Activity and the Quality Evaluation of Dioscorea opposita Thunb. Based on the Analysis of High Content Screening / 中国药学杂志

OBJECTIVE: To compare the differences of macrophages phagocytic activities between Dioscorea opposita Thunb. cv. Tiegun(abbreviated as TG) and Dioscorea opposita Thunb. but not Tiegun(abbreviated as NTG). METHODS: CCK-8 assay was used to test the cytotoxicity. On the basis of non-toxic dose, the high content screening(HCS) cell imaging analysis was applied to test the phagocytic ability of RAW264.7 cells engulfing GFP-E. coli in vitro, and the phagocytic rates between different kinds of samples at various concentrations were calculated. Furthermore, the biological potency was measured according to the qualitative response parallel method to better evaluate the difference of TG and NTG, by translating phagocytic rates into biological potency. RESULTS: At the range of 0.695 - 5.56 mg(crude drug) •mL-1, all 11 batches samples have no toxicity. The results of HCS displayed that every sample could promote cell phagocytic activity in varying degrees at a certain concentration. RAW264.7 cells could engulf the GFP-E. coli, and the images of HCS reflected the situation of phagocytosis clearly. In addition, the results of biopotency showed that the biopotency value of different samples was between 39.56 to 100, and the average biopotency value of TG was significantly higher than NTG (P < 0.05). CONCLUSION: In vitro experiments showed that all the samples could promote the phagocytic activity of RAW264.7 cells at different degrees. And the average biopotency value of TG was significantly higher than NTG, which is consistent with the saying that "TG has a better quality".

Chemical Constituents of Ethyl Acetate Fraction from Dioscorea bulbifera / 中国药学杂志

OBJECTIVE: To study the chemical constituents of ethyl acetate fraction from Dioscorea bulbifera. METHODS: This compounds were isolated and purified with silica gel and Sephadex LH-20 and preparative liquid chromatography. Their structures were determined on the basis of physicochemical properties and spectroscopic analysis. RESULTS: Twenty-four compounds were isolated and identified as ephemeranyhoquinone(1), N-methyl-2-pyrolidinone(2), aurantiamide acetate(3), 3,4-dihydroxybenzoic acid ethyl ester(4), 2, 7-dihydroxy-3, 4-dimethoxyphenanthrene(5), 4, 5-dihydroblumenol A(6), C-veratroylglycol(7), teasperol(8), vomifoliol(9), Z-6, 7-ligustilide(10), 2, 3, 7-trihydroxy-4-methoxyphenanthrene(11), p-hydroxyphenylacetic acid methyl ester(12), 3, 5, 7, 4′-tetrahydroxyflavone(13), 3, 5, 3′-trimethoxyquercetin(14), 3, 4-dihydroxybenzoic acid(15), 3, 5, 7, 3′, 4′-pentahydroxyflavone(16), daucosterol(17), 5, 3 ′, 4′-trihydroxy-3, 7-dimethoxyflavone(18), epicatechin(19), 3, 7-dihydroxy-2, 4-dimethoxy-9, 10-dihydrophenanthrene(20), methyl eucomate(21), catechins(22), 5, 7-dihydroxy-3, 4′-dimethoxyflavone(23), and 3, 4-dihydroxybenzoic acid methyl ester(24). CONCLUSION: Except compounds 11, 14, 15, 18-20, 22 other compounds are isolated from the plant for the first time.

Analysis of transcriptome of Dioscorea zingiberensis and excavation of key enzyme genes of saponin biosynthesis / 中草药

Objective To compare and analyze the transcriptome of rhizome and leaves of Dioscorea zingiberensis, and excavate the key enzyme genes related to the saponin biosynthetic pathway in D. zingiberensis. Methods The transcriptome of rhizome and leaves of D. zingiberensis were sequenced by Illumina HiSeq2000 high-throughput sequencing technique. According to sequence annotate results to find the differentially expressed genes. Then the key enzyme genes related to the biosynthesis of diosgenin were identified according to the content of saponins in rhizomes and leaves of D. zingiberensis. The expression levels of some candidate genes were analyzed by real-time fluorescence quantitative PCR (qRT-PCR). Results A total of 81 660 Unigenes were gained and 64.33% of them were annotated in NT, NR, Swiss-Prot, KOG, GO, and KEGG databases. Based on their expression and KEGG annotation, totally 227 catalytic enzyme genes of 29 kinds that may participate in D. zingiberensis saponin biosynthetic pathway were screened. The expression pattern of some catalytic enzymes was correlated with the content of saponin. Also were found five D. zingiberensis endophyte genes. Conclusion This experiment obtained candidate key enzyme genes tentatively that involved in the biosynthesis of saponin. Some candidate enzyme genes may participate in the post-modification process of steroidal saponins in D. zingiberensis. In additon, it was found that D. zingiberensis endotrophic bacteria may be involved in the saponin biosynthesis. The results laid a foundation to further elucidate the molecular mechanism of sapogenin synthesis pathway.

Chemical constituents from Dioscorea opposite / 中草药

Objective To study the chemical constituents of Dioscorea opposita. Methods The constituents were isolated and purified by silica gel, Sephadex LH-20, and ODS column chromatography. Their structures were elucidated by NMR spectra comparison with reference data. Results Fourteen compounds were isolated from 95% ethanol extract of D. opposite. All of them included eight diarylheptanoids (1-8) and six nitrogen compounds (9-14). They were elucidated by spectroscopic data as 5-ethoxy-1,7-diphenylheptan-3-one (1), 5-hydroxy-1,7-bis(4-hydroxyphenyl)-heptan-3-one (2), 1,7-diphenyl-4-hepten-3-one (3), 1,7-bis (4-hydroxyphenyl)-4-hepten-3-one (4), hannokinol (5), 1-(4-hydroxy-3-methoxyphenyl)-7-(4-hydroxyphenyl)-3,5-heptanediol (6), 1,7-bis (4-hydroxy-3-methoxyphenyl)-3,5-heptanediol (7), 1,7-bis(4-hydroxyphenyl)-1,5-epoxy-3-hydroxyheptane (8), trans-N- coumaroyltyramine (9), trans-N-feruloyltyramine (10), cis-N-coumaroyltyramine (11), trans-N-cinnamoyltyramine (12), pyrrolezanthine-6-ethyl ether (13), divaricataester A (14). Conclusion Compounds 1 is a new natural product, named as dioscoheptone A. Compounds 3, 4, 10, 12, 13, and 14 are isolated from the family of dioscoreaceae for the first time. Compounds 2, 6, and 8 are isolated from D. opposita for the first time.