Effects of gut microbiota on omega-3-mediated ovary and metabolic benefits in polycystic ovary syndrome mice.

BACKGROUND: Polycystic ovary syndrome (PCOS) is a common reproductive endocrine disorder that frequently exhibits low-grade inflammation, pro-oxidant activity, and gut dysbiosis. PCOS has become one of the leading causes of female infertility worldwide. Recently, omega-3 polyunsaturated fatty acids (PUFAs) have been proven to benefit metabolic disorders in PCOS patients. However, its roles in the regulation of metabolic and endocrinal balances in PCOS pathophysiology are not clear. In the present study, we aimed to explore how omega-3 PUFAs alleviate ovarian dysfunction and insulin resistance in mice with dehydroepiandrosterone (DHEA)-induced PCOS by modulating the gut microbiota. METHODS: We induced PCOS in female mice by injecting them with DHEA and then treated them with omega-3 PUFAs. 16S ribosomal DNA (rDNA) amplicon sequencing, fecal microbiota transplantation (FMT) and antibiotic treatment were used to evaluate the role of microbiota in the regulation of ovarian functions and insulin resistance (IR) by omega-3 PUFAs. To further investigate the mechanism of gut microbiota on omega-3-mediated ovarian and metabolic protective effects, inflammatory and oxidative stress markers in ovaries and thermogenic markers in subcutaneous and brown adipose tissues were investigated. RESULTS: We found that oral supplementation with omega-3 PUFAs ameliorates the PCOS phenotype. 16S rDNA analysis revealed that omega-3 PUFA treatment increased the abundance of beneficial bacteria in the gut, thereby alleviating DHEA-induced gut dysbiosis. Antibiotic treatment and FMT experiments further demonstrated that the mechanisms underlying omega-3 benefits likely involve direct effects on the ovary to inhibit inflammatory cytokines such as IL-1ß, TNF-α and IL-18. In addition, the gut microbiota played a key role in the improvement of adipose tissue morphology and function by decreasing multilocular cells and thermogenic markers such as Ucp1, Pgc1a, Cited and Cox8b within the subcutaneous adipose tissues. CONCLUSION: These findings indicate that omega-3 PUFAs ameliorate androgen-induced gut microbiota dysbiosis. The gut microbiota plays a key role in the regulation of omega-3-mediated IR protective effects in polycystic ovary syndrome mice. Moreover, omega-3 PUFA-regulated improvements in the ovarian dysfunction associated with PCOS likely involve direct effects on the ovary to inhibit inflammation. Our findings suggest that omega-3 supplementation may be a promising therapeutic approach for the treatment of PCOS by modulating gut microbiota and alleviating ovarian dysfunction and insulin resistance.

Analysis of proanthocyanidins and flavonols in the seedpods of Chinese Antique Lotus: A rich source of antioxidants.

Antique Lotus (Nelumbo) is a perennial aquatic plant with unique historical significance and cultural value, whereas its potential economic value hasn't been fully explored. The present study showed that lotus seedpods had significantly higher antioxidant capacity than other parts by FRAP, ABTS, and ORAC assays and analyzed the proanthocyanidins and flavonols in the seedpods of Antique Lotus. Polyphenols contributed to great antioxidant activity and 51 polyphenols were identified by UPLC-TQ-MS analysis. In which, 27 compounds were identified from lotus seedpods for the first time, including 20 trimers, 5 dimers and 2 tetramers of proanthocyanidin. Total proanthocyanidins explained 70%-90% of the different antioxidant activities and the content of proanthocyanidin trimers showed the strongest correlations with the antioxidant activities. This study provided a fundamental reference for the research of polyphenols in lotus and found that Antique Lotus seedpod extracts have the promising prospects of additives used in feed and food processing.

Phytochemical and underlying mechanism of Mikania micrantha Kunth on antibiotic resistance genes, and pathogenic microbes during chicken manure composting.

Chicken manure is a source of antibiotic resistance genes (ARGs) and pathogenic microbes. Mikania micrantha Kunth (MM) is an invasive plant containing phytochemicals as antimicrobial agents. To explore its impacts on ARGs and pathogen-host interactions (PHIs), MM was added to composting mixtures. The findings indicated that compared with control (CK), MM significantly improved the phytochemical abundances, particularly stilbenoids and diarylheptanoids (4.87%), and ubiquinones (2.66%) in the treatment (T) compost. Besides, significant ARGs reduction was noted, where rpoB2, RbpA, FosB1, vatC, and vatB were removed from T compost. PHIs significantly declined in T compost, where the growth of Xanthomonas citri, Streptococcus pneumoniae, Fusarium graminearum, Vibrio cholerae, and Xanthomonas campestris were inhibited. Multiple variable analyses demonstrated that temperature and pH revealed a significant role in ARGs and PHIs decline. Accordingly, this study considerably recommends MM as a promising compost additive in terms of its antimicrobial potential toward pathogenic microbes and ARGs.

Transcriptome analysis reveals the mechanisms of hepatic injury caused by long-term environmental exposure to atrazine in juvenile common carp (Cyprinus carpio L.).

Atrazine (ATZ) is the second most commonly used herbicide worldwide, resulting in the pollution of water bodies and affecting the economic benefits of aquaculture. ATZ is known to cause liver damage in the common carp, Cyprinus carpio L., one of the most widely cultivated fish in China, but the underlying mechanisms are poorly understood. In this study, juvenile common carp Cyprinus carpio L. were exposed to three different environmental levels (0.4, 0.8, and 1.2 µg/L) of ATZ for 12 weeks and changes in the liver transcriptomes between the high-dose group and the control group were analyzed. The data showed that different levels of ATZ exposure caused hepatotoxicity in juvenile carp, shown by biochemical parameters and histopathological changes. Comparative transcriptomics showed that high-dose ATZ exposure led to alterations in the expression of various lipid metabolism-related gene changes, including genes associated with metabolic pathways, fatty acid metabolism, and fatty acid elongation. Furthermore, a connection network analysis of the top 100 differentially expressed genes (DEGs) showed a variety of associations between high-dose ATZ-induced liver damage and the principal DEGs, indicating the complexity of hepatotoxicity induced by ATZ. In conclusion, the molecular mechanisms underlying ATZ-triggered hepatotoxicity in juvenile carp are primarily related to impaired lipid metabolism.

Microbial response mechanism of plants and zero valent iron in ecological floating bed: Synchronous nitrogen, phosphorus removal and greenhouse gas emission reduction.

Iron-based ecological floating beds (EFBs) are often used to treat the secondary effluent from wastewater treatment plant to enhance the denitrification process. However, the impact and necessity of plants on iron-based EFBs have not been systematically studied. In this research, two iron-based EFBs with and without plants (EFB-P and EFB) were performed to investigate the response of plants on nutrient removal, GHG emissions, microbial communities and functional genes. Results showed the total nitrogen and total phosphorus removal in EFB-P was 45-79% and 48-72%, respectively, while that in EFB was 31-67% and 44-57%. Meanwhile, plants could decrease CH4 emission flux (0-3.89 mg m-2 d-1) and improve CO2 absorption (4704-22321 mg m-2 d-1). Plants could increase the abundance of Nitrosospira to 1.6% which was a kind of nitrifying bacteria dominant in plant rhizosphere. Among all denitrification related genera, Simplicispira (13.08%) and Novosphingobium (6.25%) accounted for the highest proportion of plant rhizosphere and iron scrap, respectively. Anammox bacteria such as Candidatus_Brocadia was more enriched on iron scraps with the highest proportion was 1.21% in EFB-P, and 2.20% in EFB. Principal co-ordinates analysis showed that plants were the critical factor determining microbial community composition. TN removal pathways were mixotrophic denitrification and anammox in EFB-P while TP removal pathways were plant uptake and phosphorus-iron coprecipitation. In general, plants play an important directly or indirectly role in iron-based EFBs systems, which could not only improve nutrients removal, but also minimize the global warming potential and alleviate the greenhouse effect to a certain extent.

Simultaneous denitrification and iron-phosphorus precipitation driven by plant biomass coupled with iron scraps in subsurface flow constructed wetlands.

This study investigated the interaction between plant biomass and iron scraps and their influence on nitrogen (including nitrate and ammonia) and phosphorus removal in the subsurface flow constructed wetland. The results showed that with the addition of 0.5 g L-1 of plant biomass and 5.0 g L-1 of iron scraps, the nitrate, total nitrogen and total phosphorus removal were simultaneously improved. During 35 days of continuous operation, the plant biomass played main effect on the enhanced denitrification, accounting for about 57%, while iron scraps enhanced the other 43% of nitrogen removal and most phosphorus removal through precipitation inside the wetlands. Iron scraps could benefit the degradation of cellulose into low molecular carbohydrates by 10%, and the biomass could promote the oxidation of iron and increase the total phosphorus removal by 15%. Plant biomass coupled with iron scraps also improved simultaneously the richness, diversity and evenness of microbial community and promoted the abundance of Nitrospira (17.37%) and Thiobacillus (8.46%) in wetlands. In practice, putting iron scraps as matrix and placing plant biomass in the influent region would be a better choice. This research would provide a new method for effective utilization of plant biomass and iron scraps and further treatment of low-polluted wastewater in the wetlands.

Effects of Calf Blood-Deproteinized Extract Ophthalmic Gel Combined with Sodium Hyaluronate Eye Drops on Conjunctival Hyperemia Score and Tear Film Stability in Patients with Dry Eye.

Objective: The aim of this paper is to research the impact of calf blood-deproteinized extract ophthalmic gel combined with sodium hyaluronate eye drops on conjunctival hyperemia score and tear film stability in patients having dry eye. Methods: 144 patients having dry eye disease who were hospitalized from July 2018 to July 2021 were chosen as the research targets, which were composed of the control group and research group on the basis of the sequences of hospitalization, and each has 72 cases. The sodium hyaluronate eye drops were gained in the control one. In accordance with the control one, the study ones were given the combined treatment with eye drops of sodium hyaluronate. Treatment of calf blood-deproteinized extract ophthalmic gel, the scores of conjunctival hyperemia, tear film stability, visual function, curative effect, and their adverse reactions were observed and made comparisons among the two groups. Results: After the treatments of 2 weeks and later 1 month, the numerical values of the conjunctival hyperemia were fewer in the categories which had been mentioned above, and in contrast with the control one (P < 0.05), the numerical values of the conjunctival hyperemia in the study one were distinctly fewer. The data of SIt, BUT, and the heights of the central tear river of the lower eyelid were higher than those before cure. In comparison with the control one, the FL of the study one was distinctly lower, and the heights of SIt, BUT, and the central tear river of the lower eyelid were obviously better than the control one (P < 0.05). After therapy, the visual contrast sensitivity of the patients in two categories at 6.4 c/d and 12c/d increased in comparison with those before treatment, and the visual contrast sensitivity of the study group at 6.4 c/d and 12c/d was apparently higher than the data of control one (P < 0.05); the effective rate of treatment in the study one was 97.22%, which had a higher situation than the control one, 88.89% (P < 0.05); the probability of occurrence of the negative events in the study one after treatment was 4.17%, which was distinctly below the control one, 15.28% (P < 0.05). Conclusion: In order for the treatments of patients having dry eye disease, the use of calf blood-deproteinized extract ophthalmic gel having the combinations with eye drops of sodium hyaluronate can enhance the extent of visual impairment and conjunctival hyperemia with effect and improve the stability of tear film, with significant safety and high efficiency.

Culture-dependent and culture-independent methods reveal microbe-clay mineral interactions by dissimilatory iron-reducing bacteria in an integral oilfield.

Fe(III) may be reasonably considered as one of the most important electron acceptors in petroleum reservoir ecosystems. The microbial mineralization of clay minerals, especially montmorillonite, is also of great significance to the exploration of petroleum and gas reservoirs. The bioreduction mechanisms of iron-poor minerals in petroleum reservoirs have been poorly investigated. This study investigated the bioreduction of montmorillonite by dissimilatory iron-reducing bacteria (DIRB) in petroleum reservoirs based on culture-independent and culture-dependent methods. Microbial diversity analysis revealed that Halolactibacillus, Bacillus, Alkaliphilus, Shewanella, Clostridium, and Pseudomonas were the key genera involved in the bioreduction of Fe(III). Through the traditional culture-dependent method, most of the key genera were isolated from the samples collected from petroleum reservoirs. Traditional culture-dependent methods can be used to reveal the metabolic characteristics of microorganisms (such as iron-reduction efficiency) to further elucidate the roles of different species (B. subtilis and B. alkalitelluris) in the environment. Moreover, many species with high iron-reduction efficiencies and relatively low abundances in the samples, such as Tessaracoccus and Flaviflexus, were isolated from petroleum reservoirs for the first time. The combination of culture-dependent and culture-independent methods can be used to further the understanding of the microbial communities and the metabolic characteristics of DIRB in petroleum reservoirs. Structural alterations that occurred during the interactions of microorganisms and montmorillonite were revealed through scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), and X-ray powder diffraction (XRD). The physical and chemical analysis results demonstrated that microorganisms from petroleum reservoirs can dissolve iron-poor montmorillonite and promote the release of interlayer water. The secondary minerals illite and clinoptilolite were observed in bioreduced smectite. The formation of secondary minerals was closely related to the dissolution degrees of minerals based on iron reduction.

New insights on the analysis of phytosterols in pollen and anther wall of tree peony (Paeonia ostii 'Fengdan') revealed by GC-MS/MS.

Phytosterols exhibit a wide range of pharmacological activities and have no side-effects, which have attracted more and more attention. In this study, two pre-treatment methods (alkaline hydrolysis and acid-alkali hydrolysis) were performed in parallel for the first time to investigate the effects on phytosterol analysis. The aim is to develop an analytical method for phytosterols in pollen and anther wall of tree peony (Paeonia ostii 'Fengdan') by gas chromatography-mass spectrometry. The result indicated that the contents of phytosterols in pollen (3390.02 mg/100 g DW) and anther wall (929.70 mg/100 g DW) of P. ostii 'Fengdan' were relatively high. Meanwhile, eleven phytosterols were identified, among which five phytosterols were first identified in tree peony pollen. Moreover, the effects of wall breaking and different developmental stages on phytosterol contents of pollen and anther wall of P. ostii 'Fengdan' were also discussed. The result showed that wall breaking was beneficial to sterol detection of pollen, but not necessary for anther wall. For the best harvest time, the stage of S2 had the highest content of phytosterols. In conclusion, this study successfully establishes an analytical method for phytosterols by gas chromatography-mass spectrometry, and lays a foundation for the development and utilization of pollen and anther wall of P. ostii 'Fengdan' in functional foods and pharmaceutical industry.

The chromosome-scale genome assembly, annotation and evolution of Rhododendron henanense subsp. lingbaoense.

Rhododendron henanense subsp. lingbaoense (hereafter referred to as R. henanense) is an endemic species naturally distributed in the Henan province, China, with high horticultural, ornamental and medicinal value. Herein, we report a de novo genome assembly for R. henanense using a combination of PacBio long read and Illumina short read sequencing technologies. In total, we assembled 634.07 Mb with a contig N50 of 2.5 Mb, representing ~96.93% of the estimated genome size. By applying Hi-C data, 13 pseudochromosomes of R. henanense genome were assembled, covering ~98.21% of the genome assembly. The genome was composed of ~65.76% repetitive sequences and 31,098 protein-coding genes, 88.77% of which could be functionally annotated. Rhododendron henanense displayed a high level of synteny with other Rhododendron species from the Hymenanthes subgenus. Our data also suggests that R. henanense genes related to stress responses have undergone expansion, which may underly the unique abiotic and biotic stress resistance of the species. This alpine Rhododendron chromosome-scale genome assembly provides fundamental molecular resources for germplasm conservation, breeding efforts, evolutionary studies, and elucidating the unique biological characteristics of R. henanense.

Pilot-Scale Airlift Bioreactor with Function-Enhanced Microbes for the Reduction of Refinery Excess Sludge.

A pilot-scale airlift bioreactor (ALBR) system was built and operated continuously for refinery excess sludge (RES) reduction. Combined ALBR and function-enhanced microbes (composed of photosynthetic bacteria and yeast) were integrated into the system. The pilot-scale ALBR was operated for 62 days, and the start-up time was 7 d. Continuous operation showed that the sludge reduction efficiency was more than 56.22%, and the water quality of the effluent was satisfactory. This study focused on investigating the effects of hydraulic retention time (HRT) on the stability of the system and the effect of sludge reduction. Under different HRT conditions of 40, 26.7, 20, and 16 h, the sludge reduction rates reached 56.22%, 73.24%, 74.09%, and 69.64%, respectively. The removal rates of chemical oxygen demand (COD) and total nitrogen (TN) decreased with decreasing HRT, whereas the removal rate of NH4+-N increased. The removal rate of total phosphorus (TP) was approximately 30%. Results indicate that the ALBR and function-enhanced microbe system can reduce sludge and treat sewage simultaneously, and the effluent is up to the national emission standard. Addition of function-enhanced microbes can promote the degradation of petroleum hydrocarbon substances in the sludge, especially alkanes with low carbon numbers. This study suggests that the optimal HRT for the system is 16 h. The total operation cost of the ALBR combined with the function-enhanced microbe system can be reduced by 50% compared with the cost of direct treatment of the RES system.

Preparative separation of five polyphenols from the fruits of Sorbus pohuashanensis Hedl. by high-speed counter-current chromatography.

The fruits of Sorbus pohuashanensis Hedl. (S. pohuashanensis) are rich in polyphenols with many beneficial effects such as anti-inflammatory, anti-tussive, anti-asthmatic and anti-cancer. In this study, five polyphenols, including three phenolic acids and two flavonoids, were successfully prepared from the fruits of S. pohuashanensis by high-speed counter-current chromatography (HSCCC) using different solvent systems for the first time. Ethyl acetate-n-butanol-water (3.5:1.5:5, v/v) was screened as the two-phase system to separate neochlorogenic acid (1), chlorogenic acid (2), quercetin 3-O-(6″-α-L-rhamnopyranosyl-4'″-α-L-rhamnopyranosyl)-ß-D-glucopyranoside (3) and rutin (5). N-hexane-ethyl acetate-methanol-water (1:3:1:3.5, v/v) was first utilized to isolate 3,5-O-dicaffeoylquinic acid (4). The purities of all these compounds were above 95%. In addition, their chemical structures were identified by mass spectrometer (MS), nuclear magnetic resonance (NMR) or the standards. These results indicated that HSCCC was an effective method to separate polyphenols compounds from the fruits of S. pohuashanensis.

Biological iron nitrogen cycle in ecological floating bed: Nitrogen removal improvement and nitrous oxide emission reduction.

Ecological floating beds (EFBs) have become a superior method for treating secondary effluent from wastewater treatment plant. However, insufficient electron donor limited its denitrification efficiency. Iron scraps from lathe cutting waste consist of more than 95% iron could be used as electron donors to enhance denitrification. In this study, EFBs with and without iron scraps supplementation (EFB-Fe and EFB, respectively) were conducted to explore the impacts of iron scraps addition on nitrogen removal, nitrous oxide (N2O) emissions and microbial communities. Results showed the total nitrogen (TN) removal in EFB-Fe improved to 79% while that in EFB was 56%. N2O emission was 0-6.20 mg m-2 d-1 (EFB-Fe) and 1.74-15.2 mg m-2 d-1 (EFB). Iron scraps could not only improve nitrogen removal efficiency, but also reduce N2O emissions. In addition, high-throughput sequencing analysis revealed that adding iron scraps could improve the sum of denitrification related genera, among which Novosphingobium accounted for the highest proportion (6.75% of PFe1, 4.24% of PFe2, 3.18% of PFe3). Iron-oxidizing bacteria and iron-respiring bacteria associated with and nitrate reducing bacteria mainly concentrated on the surface of iron scraps. Principal co-ordinates analysis (PCoA) indicated that iron scraps were the key factor affecting microbial community composition. The mechanism of iron scraps enhanced nitrogen removal was realized by enhanced biological denitrification process. Iron release dynamic from iron scraps was detected in bench-scale experiment and the electron transfer mechanism was that Fe0 transferred electrons directly to NO3--N, and biological iron nitrogen cycle occurred in EFB-Fe without secondary pollution.

Comparison of Traditional Chinese Exercises and Nontraditional Chinese Exercise Modalities on Cognitive and Executive Function in Community Middle-Aged and Older Adults: A Cross-Sectional Study.

BACKGROUND: Current evidence indicates that regular exercise can have a positive impact on cognitive function in older adults, but whether different exercise modalities may induce differential protective effects in different cognitive domains is uncertain. OBJECTIVE: To compare the effect of traditional Chinese exercise (TCE) modalities and non-traditional Chinese exercise (non-TCE) modalities on cognitive and executive function in community middle-aged and older adults through a cross-sectional study. METHODS: A total of 350 community middle-aged and older adults aged over 55 years participated in this study. Information on demographic characteristics, lifestyle and behavioural habits, and regular exercise was collected by a self-designed questionnaire. Global cognitive ability and executive function were assessed using the Montreal Cognitive Assessment (MoCA) scale, the clock drawing test (CDT), the animal naming test (ANT), and the trail making test (TMT). Eligible subjects were categorized into the no regular exercise (no-RE), non-TCE, or TCE groups according to their self-reported exercise information. Comparisons of global cognitive and executive function among the three groups were conducted using ANOVA or the general linear model with adjustment for potential confounding factors. RESULTS: The results showed that for the non-TCE or TCE groups, the MoCA and CDT scores were significantly higher, and the TMT-A test time was significantly shorter than those in the no-RE group (all P < 0.05), but no significant difference was observed for the TMT-B and ANT tests. After adjustment for potential confounding factors, the MoCA, the CDT, and TMT-A scores in the TCE group were significant compared to those in the no-RE group. In addition, subgroup analysis showed that in the TCE group, the MoCA scores were significantly higher than those in the non-TCE group. Furthermore, in the TCE group, the CDT scores for those with an exercise duration of <5 years were higher and the TMT-A test time for those with an exercise duration of ≥5 years was shorter than those in the non-TCE group. CONCLUSIONS: Both TCE and non-TCE have potential protective effects on global cognitive and executive function in community middle-aged and older adults. Compared to the non-TCE modality, the TCE modality may have a more positive association with these protective effects. Furthermore, prospective studies are needed to confirm these findings.

Therapeutic manipulation of gut microbiota by polysaccharides of Wolfiporia cocos reveals the contribution of the gut fungi-induced PGE2 to alcoholic hepatic steatosis.

Alcohol abuse and alcoholic liver diseases (ALD) have been worldwide spread. Chronic alcoholism-induced overgrowth of intestinal bacteria and fungi together with the enteric dysbiosis are important pathogenic mechanisms in ALD. We demonstrated that the water-insoluble polysaccharides (WIP) from Wolfporia cocos effectively ameliorated the hepatic inflammatory injury and fat accumulation through modulating gut microbiota in mice with alcoholic hepatic steatosis (AHS). Oral administration of WIP significantly enhanced the ratio of Firmictues to Proteobacteria, increased the abundance of Lachnospiraceae including Ruminoclostridum and unidentified_clostridials, and inhibited the ethanol-induced fungal overgrowth. Treatment with WIP activated the PPAR-γ signaling and reduced the inflammation in the colonic epithelia cell, facilitating a hypoxic state that suppresses the overgrowth of fungi and Proteobacteria in the gut. In addition, we found an overwhelming increase of the commensal fungus Meyerozyma guilliermondii in the feces of mice with AHS by culturing and ITS sequencing. Inoculation of M. guilliermondii into fungi-free mice aggravated the features of AHS. M. guilliermondii was found to generate PGE2 by biotransformation of arachidonic acid. Furthermore, the gut fungi (M. guilliermondii)-induced PGE2 production in the liver was confirmed as one of the mechanisms in the chronic AHS. The current study supports the manipulation of the gut microbiota (bacteria and fungi) as an effective and alternative strategy for alleviating ALD.

Directional culture of petroleum hydrocarbon degrading bacteria for enhancing crude oil recovery.

An oxygen-constrained system of crude oil reservoir environment was constructed to stimulate the growth of indigenous microbes, such as petroleum hydrocarbon-degrading bacteria. Addition of nitrogen and phosphorus sources was investigated for the growth of petroleum hydrocarbon-degrading bacteria. The results show that nitrates and phosphates stimulated the growth of the bacteria and promoted the biodegradation of crude oil as the sole carbon source in this process. The minimum surface tension was 29.63 mN/m when the amounts of the nitrogen (NaNO3: [Formula: see text]  = 2:1) and phosphorus (KH2PO4: NaH2PO4 = 5:2) sources added were 0.8 wt% and 1.4 wt%, respectively. Furthermore, the dominant petroleum hydrocarbon-degrading bacteria were shifted from Arcobacter in production water to Pseudomonas after the first subculture and then to Bacillus after the sixth subculture. The heteroatom groups in the crude oil were biodegraded simultaneously with normal alkanes and alkyl cyclohexanes. Addition of the nutrients resulted in microbial growth, microbial community shift, and enhanced microbial degradation.

Stimulation of indigenous microbes by optimizing the water cut in low permeability reservoirs for green and enhanced oil recovery.

Low permeability oil reservoirs are a widespread petroleum reservoir type all over the world. Therefore, methods to recover these reservoirs efficiently are of importance to guarantee energy supply. Here we report our novel stimulation of indigenous microbes by optimizing the water cut in low permeability reservoirs for green and enhanced oil recovery. We aimed to investigate the characteristics of indigenous bacterial communities with changes in water cut in reservoirs by high-throughput sequencing technology, and reveal the mechanism and characteristics of the crude oil biotreatment under different crude oil-water ratio conditions and the optimum activation time of indigenous functional microbial groups in reservoirs. The indigenous microbial metabolism products were characterized by gas chromatography mass spectrometry. Results showed that Acinetobacter (47.1%) and Pseudomones (19.8%) were the main functional genus of crude oil degradation at the optimal activation time, and can reduce the viscosity of crude oil from 8.33 to 5.75 mPa·s. The dominant bacteria genus for oil recovery after activation of the production fluids was similar to those in the reservoirs with water cut of 60-80%. Furthermore seven mechanism pathways of enhancing oil recovery by the synergistic of functional microbial groups and their metabolites under different water cut conditions in low permeability reservoirs have been established.

Accurate discrimination of Gastrodia elata from different geographical origins using high-performance liquid chromatography fingerprint combined with boosting partial least-squares discriminant analysis.

Gastrodia elata from different geographical origins varies in quality and pharmacological activity. This study focused on the classification and identification of Gastrodia elata from six producing areas using high-performance liquid chromatography fingerprint combined with boosting partial least-squares discriminant analysis. Before recognition analysis, a principal component analysis was applied to ascertain the discrimination possibility with high-performance liquid chromatography fingerprints. And then, boosting partial least-squares discriminant analysis and conventional partial least-squares discriminant analysis were applied in this study. Experimental results indicated that the adaptive iteratively reweighted penalized least-squares algorithm could eliminate the baseline drift of high-performance liquid chromatography chromatograms effectively. And compared with partial least-squares discriminant analysis, the total recognition rates using high-performance liquid chromatography fingerprint combined with boosting partial least-squares discriminant analysis for the calibration sets and prediction sets were improved from 94 to 100% and 86 to 97%, respectively. In conclusion, high-performance liquid chromatography combined with boosting partial least-squares discriminant analysis, which has such advantages as effective, specific, accurate, non-polluting, has an edge for discrimination of traditional Chinese medicine from different geographical origins. And the proposed methodology is a useful tool to classify and identify Gastrodia elata from different geographical origins.

The application of combined 1H NMR-based metabolomics and transcriptomics techniques to explore phenolic acid biosynthesis in Salvia miltiorrhiza Bunge.

Salvia miltiorrhiza Bunge is a traditional Chinese medicine, and its water-soluble phenolic acid active compounds have very important medicinal value; however, the synthesis pathways of the main active ingredients remain unknown. Here, we employed nuclear magnetic resonance (NMR)-based metabolomics and transcriptomics techniques to study the biosynthesis mechanism of salvianolic acids. High-performance liquid chromatography (HPLC) combined with NMR showed an improvement over traditional techniques, and 54 metabolites were detected. The results of the multivariate statistical analysis showed that salvianolic acid B (SAB), rosmarinic acid (RA), caffeic acid, succinate, and citrate were among the multiple compounds that were increased in the methyl jasmonate (MeJA)-elicited group; the levels of sucrose, fructose, glutamine, and tyrosine were decreased. Combined with the differentially expressed genes (DEGs) found by transcriptome sequencing, we speculate that the synthesis of RA after MeJA treatment mostly occurred through caffeic acid and bypassed 4-hydroxyphenyllactic acid. This provides useful information for the study of salvianolic acids synthesis.

An insoluble polysaccharide from the sclerotium of Poria cocos improves hyperglycemia, hyperlipidemia and hepatic steatosis in ob/ob mice via modulation of gut microbiota.

Metabolic syndrome characterized by obesity, hyperglycemia and liver steatosis is becoming prevalent all over the world. Herein, a water insoluble polysaccharide (WIP) was isolated and identified from the sclerotium of Poria cocos, a widely used Traditional Chinese Medicine. WIP was confirmed to be a (1-3)-ß-D-glucan with an average Mw of 4.486 × 106 Da by NMR and SEC-RI-MALLS analyses. Furthermore, oral treatment with WIP from P. cocos significantly improved glucose and lipid metabolism and alleviated hepatic steatosis in ob/ob mice. 16S DNA sequencing analysis of cecum content from WIP-treated mice indicated the increase of butyrate-producing bacteria Lachnospiracea, Clostridium. It was also observed that WIP treatment elevated the level of butyrate in gut, improved the gut mucosal integrity and activated the intestinal PPAR-γ pathway. Fecal transplantation experiments definitely confirmed the causative role of gut microbiota in mediating the benefits of WIP. It is the first report that the water insoluble polysaccharide from the sclerotium of P. cocos modulates gut microbiota to improve hyperglycemia and hyperlipidemia. Thereby, WIP from P. cocos, as a prebiotic, has the potential for the prevention or cure of metabolic diseases and may elucidate new mechanism for the efficacies of this traditional herbal medicine on the regulation of lipid and glucose metabolism.