Multigene Phylogeny, Diversity and Antimicrobial Potential of Endophytic Sordariomycetes From Rosa roxburghii.

Rosa roxburghii Tratt. is widely applied in food, cosmetics, and traditional medicine, and has been demonstrated to possess diverse bioactivities. Plant endophytic fungi are important microbial resources with great potential for application in many fields. They not only establish mutualistic symbiosis with host plants but also produce a variety of bioactive compounds. Therefore, in the present study, endophytic fungi were isolated from R. roxburghii, the diversity and antimicrobial activities were evaluated. As a result, 242 strains of endophytic Sordariomycetes were successfully isolated. Multigene phylogenetic analyses showed that these isolates included eight orders, 19 families, 33 genera. The dominant genera were Diaporthe (31.4%), Fusarium (14.4%), Chaetomium (7.9%), Dactylonectria (7.0%), Graphium (4.5%), Colletotrichum (4.1%), and Clonostachys (4.1%). For different tissues of R. roxburghii, alpha diversity analysis revealed that the diversity of fungal communities decreased in the order of root, fruit, stem, flower, leaf, and seed, and Clonostachys and Dactylonectria exhibited obvious tissue specificity. Meanwhile, functional annotation of 33 genera indicated that some fungi have multitrophic lifestyles combining endophytic, pathogenic, and saprophytic behavior. Additionally, antimicrobial activities of endophytic Sordariomycetes against Lasiodiplodia theobromae, Botryosphaeria dothidea, Colletotrichum capsici, Pyricularia oryzae, Rhizoctonia solani, Fusarium oxysporum, Pseudomonas syringae, Pantoea agglomerans, Staphylococcus aureus, Bacillus subtilis, Escherichia coli, and Pseudomonas aeruginosa were screened. Dual culture test assays showed that there were 40 different endophytic species with strong inhibition of at least one or moderate inhibition of two or more against the 12 tested strains. The results from the filter paper diffusion method suggested that extracellular metabolites may be more advantageous than intracellular metabolites in the development of antimicrobial agents. Eleven isolates with good activities were screened. In particular, Hypomontagnella monticulosa HGUP194009 and Nigrospora sphaerica HGUP191020 have shown promise in both broad-spectrum and intensity. Finally, some fungi that commonly cause disease have been observed to have beneficial biological activities as endophytic fungi. In conclusion, this study showed the species composition, alpha diversity, and lifestyle diversity of endophytic Sordariomycetes from R. roxburghii and demonstrated these isolates are potential sources for exploring antimicrobial agents.

Changes in Intestinal Microbiota of Type 2 Diabetes in Mice in Response to Dietary Supplementation With Instant Tea or Matcha.

OBJECTIVE: Gut microbiota plays a key role in metabolism and health in diabetes patients with gastrointestinal microbiota dysbiosis. Thus, regulating the ecological balance of gut microbiota may provide a pathway toward improvement for these patients. Our previous study showed that functional ingredients in tea may inhibit cornstarch digestion in vitro. METHODS: A cornstarch-tea diet was developed, and in this study we investigated the effects of such a diet on blood glucose and gut microbiota in diabetic mice. RESULTS: Diabetes resulted in significant weight loss, hyperphagia and hyperglycemia. 16S rDNA sequencing revealed that in diabetes there is significantly increased Bacteroidaceae, Helicobacteraceae, Ruminococcaceae, Enterobacteriaceae, Rikenellaceae and Saccharibacteria_genera_incertae_sedis, and significantly decreased Lactobacillaceae, Prevotellaceae, Coriobacteriaceae, Verrucomicrobiaceae and Bifidobacteriaceae. The cornstarch‒tea diet resulted in a trend toward reduced blood glucose, with particularly increased levels of Coriobacteriaceae, Lactobacillaceae, Prevotellaceae and Bifidobacteriaceae, and decreased Bacteroidaceae, Ruminococcaceae, Helicobacteraceae and Enterobacteriaceae. CONCLUSIONS: Instant tea and matcha supplementation had beneficial effects on regulation of blood glucose and gut microbiota, reversing the changes in microbiota caused by alloxan injection. The cornstarch‒tea regulation pathway is involved in bacterium group regulation rather than single-species regulation, which suggests that cornstarch combined with tea may be used as a functional food supplement for diabetes patients.

[Effect of transcutaneous neuromuscular electrical stimulation on prevention of intensive care unit-acquired weakness in chronic obstructive pulmonary disease patients with mechanical ventilation].

OBJECTIVE: To evaluate the effect of transcutaneous neuromuscular electrical stimulation on prevention of intensive care unit-acquired weakness (ICU-AW) in chronic obstructive pulmonary disease (COPD) patients with mechanical ventilation. METHODS: A prospective randomized controlled study was conducted. Sixty COPD patients aged 18-85 years old who were accepted mechanical ventilation therapy admitted to general intensive care unit (ICU) of the First Affiliated Hospital of Hunan University of Medicine from October 2017 to October 2018 were enrolled. Patients were divided into control group (n = 30) and intervention group (n = 30) by random number table method. All patients were accepted routine treatment, and on this basis, the intervention group was applied transcutaneous neuromuscular electrical stimulation on the extremities (twice a day, 30 minutes each time) after 24 hours of admission until ICU discharge. The Medical Research Council muscle strength score (MRC-Score), grip strength, incidence of ICU-AW on the 7th day after admission and on the day of ICU discharge; modified Barthel index score on the day of ICU discharge; and duration of mechanical ventilation, the length of ICU stay, and the length of hospital stay were compared between the two groups. RESULTS: Twenty-nine and 27 patients in the control group and the intervention group respectively finally completed the study in dividually. There was no significant difference in gender, age, Barthel index score before 2 weeks of ICU admission, body mass index or acute physiology and chronic health evaluation (APACHE) in ICU between the two groups. There was no significant difference in the MRC-Score, grip strength or incidence of ICU-AW on the 7th day after ICU admission between the two groups. Compared to the control group, the MRC-Score, grip strength and Barthel index score in the intervention group were significantly increased [MRC-Score: 55.97±8.43 vs. 46.32±7.36, grip strength (kg): 33.46±11.62 vs. 27.42±9.64, Barthel index score: 46.04±5.46 vs. 42.13±3.32, all P < 0.05], the incidence rate of ICU-AW was significantly decreased [7.4% (2/27) vs. 31.0% (9/29), P < 0.05], and duration of mechanical ventilation, the length of ICU stay, the length of hospital stay were significantly shortened [duration of mechanical ventilation (days): 5.12±2.01 vs. 7.24±4.35, the length of ICU stay (days): 8.34±2.36 vs. 10.45±2.62, the length of hospital stay (days): 13.21±2.21 vs. 15.38±3.67, all P < 0.05]. CONCLUSIONS: Transcutaneous neuromuscular electrical stimulation can effectively improve the muscle strength of COPD patients with mechanical ventilation and reduce the incidence of ICU-AW.

Chemical profiling of Banxia-Baizhu-Tianma decoction by ultra-fast liquid chromatography with tandem mass spectrometry.

Banxia-Baizhu-Tianma decoction (BBTD) is a compound formulae of traditional Chinese medicine (TCM), which has been clinically used for treatments of neural vertigo, hypertension and epilepsy with a long history. In this study, with an ultra-fast liquid chromatography coupled with quadrupole time of flight mass spectrometry (UFLC-Q-TOF-MS) method, a total of 88 components in BBTD were identified by the accurate masses and fragmentation pathways including 19 flavonoids, 8 lactones, 12 triterpenoids, 10 phenolics, 14 amino acids, 13 nucleobases and nucleosides, 7 organic acids, and 5 other compounds. In addition, under the same chromatographic conditions, we developed an ultra-fast liquid chromatography coupled with quadrupole linear ion trap mass spectrometry (UFLC-Q-TRAP-MS) method to simultaneously quantify 20 bioactive components in multiple-reaction monitoring (MRM) mode. The assay method was validated in terms of linearity, precision, repeatability, recovery and was successfully applied for determination of 12 batches of BBTD. We hope that this study work would help to reveal the chemical profiling and provide a valuable and reliable approach for quality evaluation and even efficacy material basis study of BBTD.

Optimal Extraction Study of Gastrodin-Type Components from Gastrodia Elata Tubers by Response Surface Design with Integrated Phytochemical and Bioactivity Evaluation.

Gastrodia elata tuber (GET) is a popular traditional Chinese medicines (TCMs). In this study, response surface methodology (RSM) with a Box⁻Behnken design (BBD) was performed to optimize the extraction parameters of gastrodin-type components (gastrodin, gastrodigenin, parishin A, parishin B, parishin C and parishin E). Different from the conventional studies that merely focused on the contents of phytochemical, we gave consideration to both quantitative analysis of the above six components by HPLC and representative bioactivities of GET, including antioxidation and protection of human umbilical vein endothelial cells (HUVEC). Four independent variables (ethanol concentration, liquid-material ratio, soaking time and extraction time) were investigated with the integrated evaluation index of phytochemical contents. With the validation experiments, the optimal extraction parameters were as follows: ethanol concentration of 41%, liquid⁻solid ratio of 28.58 mL/g, soaking time of 23.91 h and extraction time of 46.60 min. Under the optimum conditions, the actual standardized comprehensive score was 1.8134 ± 0.0110, which was in accordance with the predicted score of 1.8100. This firstly established method was proved to be feasible and reliable to optimize the extraction parameters of the bioactive components from GET. Furthermore, it provides some reference for the quality control and extraction optimization of TCMs.

Comparative analysis of sixteen active compounds and antioxidant and anti-influenza properties of Gardenia jasminoides fruits at different times and application to the determination of the appropriate harvest period with hierarchical cluster analysis.

ETHNOPHARMACOLOGICAL RELEVANCE: Gardenia jasminoides fruit (GJF) is used as a well-known traditional folk medicine, a food and a natural colorant in Asia with a long history. The herbal medicine has usually been harvested in the autumn from September to November. However, this time span is too long and might result in the quality instability of GJF. AIM OF STUDY: We aimed to conduct the comprehensive quality evaluation of GJF including the quantitative analysis of the bioactive components and the main bioactivities, and further to determine the most appropriate harvest time of this phytomedicine. MATERIALS AND METHODS: In this study, an UFLC-Q-TRAP-MS/MS method was established to quantify 7 iridoid glycosides (geniposide, geniposidic acid, secoxyloganin, gardenoside, genipin 1-gentiobioside, scandoside methyl ester, and shanzhiside), 7 phenylpropanoid acids (chlorogenic acid, cryptochlorogenic acid, neochlorogenic acid, isochlorogenic acid A, isochlorogenic acid B, isochlorogenic acid C, and caffeic acid) and 2 carotenoids (crocin-1 and crocin-2) in GJF. With this method, nine samples of GJF harvested at different times were analyzed and compared. These samples were also investigated and compared in terms of their antioxidant activity (DPPH free radical scavenging, ABTS free radical scavenging, ferric-reducing antioxidation) and anti-influenza activity (neuraminidase inhibition), which are closely related to the GJF efficacies. Then, hierarchical cluster analysis (HCA) was separately performed for the quantitative analysis and bioactivity evaluation in vitro. RESULTS: The HCA results demonstrated that three GJF samples (S5, S6, and S7) were clustered into one group for both quantitative analysis and bioactivity evaluation in vitro; these three samples were found to have the highest standardized scores in both the former (12.775, 12.106, 10.817) and the latter (3.406, 3.374, 3.440). Based on the comprehensive results, the optimum harvest period was confirmed to extend from mid-October to early-November. CONCLUSIONS: This study firstly validated the use of UFLC-Q-TRAP-MS/MS method for the determination of 16 bioactive components in GJF. It was also the first time that a quantitative analysis and a bioactivity assay in vitro were integrated for the determination of the most appropriate harvest period of GJF. We hope this paper may provide some reference to studies of appropriate harvest periods and even the quality control of TCMs.

Effect of tea products on the in vitro enzymatic digestibility of starch.

The importance of postprandial hyperglycemia in the treatment of diabetes has been recognized recently. Tea products, such as tea polyphenols (TP), epigallocatechin gallate (EGCG), matcha, and instant tea, were chosen as constituents of tea-flour food, aimed at regulating the release of glucose from starchy foods in the postprandial period. Six starches were chosen for internal composition analysis and hydrolysis studies in vitro. Corn starch, wheat starch, and lily root flour appeared to have higher resistant starch content, slower digestion profiles, and lower kinetic constants, implying sustained release of glucose in the gastrointestinal tract. The effect of tea products on starch digestion was determined in order to get a desired formulation of dietary product for patients with hyperglycemia. Compared with macha and instant tea, TP and EGCG exerted greater inhibition of amylase and amyloglucosidase, especially for corn starch with 0.5% TP or 0.5% EGCG.

Influence of brewing conditions on taste components in Fuding white tea infusions.

BACKGROUND: White tea has received increasing attention of late as a result of its sweet taste and health benefits. During the brewing of white tea, many factors may affect the nutritional and sensory quality of the resulting infusions. The present study aimed to investigate the effect of various infusion conditions on the taste components of Fuding white tea, including infusion time, ratio of tea and water, number of brewing steps, and temperature. RESULTS: Brewing conditions had a strong effect on the taste compound profile and sensory characteristics. The catechin, caffeine, theanine and free amino acid contents generally increased with increasing infusion time and temperature. Conditions comprising an infusion time of 7 min, a brewing temperature of 100 °C, a tea and water ratio of 1:30 or 1:40, and a second brewing step, respectively, were shown to obtain the highest contents of most compounds. Regarding tea sensory evaluation, conditions comprising an infusion time of 3 min, a brewing temperature of 100 °C, a tea and water ratio of 1:50, and a first brewing step, resulted in the highest sensory score for comprehensive behavior of color, aroma and taste. CONCLUSION: The results of the present study reveal differences in the contents of various taste compounds, including catechins, caffeine, theanine and free amino acids, with respect to different brewing conditions, and sensory scores also varied with brewing conditions. © 2016 Society of Chemical Industry.

Structure and bioactivity of a polysaccharide extracted from protocorm-like bodies of Dendrobium huoshanense.

The crude polysaccharides of Dendrobium huoshanense were fractionated by anion-exchange chromatography and gel permeation chromatography, giving one homogeneous fraction DHP-4A with molecular weight of 2.32 × 10(5)Da. UV spectrum indicated that there was no existence of proteins and nucleic acids in DHP-4A. Monosaccharide analysis revealed that DHP-4A was made up of glucose, arabinose, mannose and rhamnose with a molar ratio of 13.8:3.0:6.1:2.1. The backbone of DHP-4A consisted of (1 → 6)-linked glucose, (1 → 6)-linked mannose and (1 → 3,6)-linked mannose. The ßL-Rhap-(1 → 2)-ß-L-Rhap-(1 → 4)-ß-D-Manp-(1 → and α-L-Araf-(1 → 3)- α-L-Araf -(1 → 3)-α-L-Araf-(1 → were regarded as the branches attached to the C-3 position of (1 → 6)-linked mannose in the backbone. The sugar residue sequence was further determined by NMR spectra including (1)H NMR, (13)C NMR, HSQC and HMBC. Pharmacological tests showed that DHP-4A can significantly stimulate RAW 264.7 macrophage cells to secrete NO, TNF-α, IL-6 and IL-10 via activation of p38, ERK, JNK and translocation of nuclear NF-κB, indicating this polysaccharide possesses good immunoregulatory activity.

[Study on antioxidant activity of constituents from mulberry leaf].

OBJECTIVE: To study the chemical constituents with antioxidant activity from mulberry leaf. METHODS: Siliga gel column chromatography was used to isolate, and their structures were identified by spectras. METHODS: Isolated constituents were tested in vitro for antioxidant activity against 1, 1-diphenyl -2-picrylhydrazyl (DPPH) and 2, 2' -azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) (ABTS+) radicals. RESULTS: Four known flavonoids were isolated from mulberry leaf and their structures were identified as kaempferol-3-O-beta-D-glucopyranoside (I), quercetin-3-O-beta-D-glucopyranoside (II), quercitrin (III), morin-3-O-beta-D-glucopyranoside (IV). All these compounds showed DPPH and ABTS+ radicals scavenging activity. CONCLUSION: Compound IV has the strongest ABTS+ radicals scavenging effects.