Efficacy and mechanism of action of Yanxiao Di'naer formula for non-alcoholic steatohepatitis treatment based on metabolomics and RNA sequencing.

ETHNOPHARMACOLOGICAL RELEVANCE: Nonalcoholic fatty liver disease (NAFLD) is the most prevalent chronic liver disease worldwide. Nonalcoholic steatohepatitis (NASH) is a crucial component of this disease spectrum. The Yanxiao Di'naer formula (YXDNE) is an Uyghur medical extract that has been used in folk medicine to treat hepatitis for a long time. However, the role and mechanism of action of YXDNE in NASH treatment remains unclear OBJECTIVE: The objective of this study was to assess the effectiveness of YXDNE in treating NASH induced by injections of carbon tetrachloride combined with a high-fat high-cholesterol diet (HFHCD), and to clarify the underlying mechanisms METHODS: The compounds in the YXDNE extract were analysed for classification and proportions using ultra-performance liquid chromatography-mass spectrometry. The efficacy of YXDNE in treating abnormal lipid metabolism was evaluated in L02 cells in vitro. In addition, a C57BL/6 mouse model of NASH was established to evaluate the therapeutic efficacy of YXDNE in vivo. Metabolomics and RNA sequencing were used to analyse the therapeutic effects of YXDNE on the liver. The corresponding signalling pathways were found to target AMPKα1, PPARα, and NF-κB. The efficacy of YXDNE was validated using inhibitors or silencing RNA (siRNA) against AMPKα1 and PPARα RESULTS: This study confirmed that YXDNE treatment ameliorated NASH in a murine model of this disease. Metabolomics analysis suggested that YXDNE efficacy was associated with fatty acid catabolism and AMPK signalling pathways. RNA sequencing results showed that YXDNE efficacy in treating NASH was highly correlated with the AMPK, PPARα and NF-κB pathways. Both in vitro and in vivo experimental data demonstrated that YXDNE affected the expression of p-AMPKα1, PPARα, p-NF-κB, IκB, and p-IκB. The efficacy of YXDNE in treating NASH in vitro was cancelled when AMPK was inhibited with Compound C or PPARα was modulated via siRNA CONCLUSIONS: YXDNE may have a therapeutic effect on abnormal lipid metabolism in L02 cells and in a murine model of NASH by affecting the AMPKα1/PPARα/NF-κB signalling pathway. Therefore, YXDNE has the potential for clinical application in the prevention and treatment of NASH.

Three spirostanol saponins and a flavane-O-glucoside from the fresh rhizomes of Tupistra chinensis.

Four new compounds, including three new spirostanol saponins [tupistroside G-I (1-3)] and a new flavane-O-glucoside [tupichiside A (4)], together with ten known compounds, were isolated from the fresh rhizomes of Tupistra chinensis. The structures of the new compounds were elucidated by spectroscopic analysis and chemical evidence. All compounds were tested in vitro for their cytotoxic activities against the Human LoVo and BGC-823 cell lines, and six of them were found to possess potent cytotoxicity. Compounds 2, 8 and 9 showed significant cytotoxicity against the tested tumor cell lines with IC50 values ranging from 0.2 to 0.9µM.

Cytotoxic activity of lignans from Justicia procumbens.

Three new lignans, Pronaphthalide A (1), Procumbiene (2), and Procumbenoside J (3), along with a novel natural product Juspurpudin (4), and twelve other known lignans were isolated from Justicia procumbens. The structures of the new compounds were elucidated by extensive spectroscopic analyses and the data of 3 provided insight into the conformational equilibria existing in it. All compounds were evaluated for their in vitro cytotoxic activity against Human LoVo and BGC-823 cell lines except for compound 2, and eight of them were found to possess potent cytotoxicity. The structure-activity relationship (SAR) analysis revealed that (i) the parent structure of 2-carbonyl arylnaphthalide lactone attached with 6 and 7-OMe was the essential element; (ii) the polarity of substituents on C-4 might significantly affect the activity; (iii) a proper cyclic lipophilic group at the C-3″ and C-5″ of apiofuranose on C-4 might enhance the activity, which could optimize the application of 3 similar to VP-16.

Two new coumarins from the seeds of Solanum indicum.

Two new coumarins, (E)-2-(4-hydroxy-3-methoxybenzylidene)-5-methoxy-2H-[1,4]dioxino[2,3-h]chromene-3,9-dione (indicumin E, 1) and 7-hydroxy-6,8-dimethoxy-3-(4'-hydroxy-3'-methoxyphenyl)-coumarin (2), together with two known coumarins isofraxidin (3) and fraxetin (4), were isolated from the Solanum indicum seeds. Their structures were established on the basis of 1D and 2D spectroscopic data. Compound 1 was the rarest coumarinolignoid known to date.

Three new germacrane-type sesquiterpenes with NGF-potentiating activity from Valeriana officinalis var. latiofolia.

Three new germacrane-type sesquiterpenoids, volvalerenal F (1), volvalerenal G (2) and volvalerenic acid D (3), along with five known compounds 4-8, were isolated from the CHCl3 soluble partition of the ethanol extract of Valeriana officinalis var. latiofolia. The structures of the new compounds were determined on the basis of spectroscopic evidence, including their 1D- and 2D-NMR spectra, as well as mass spectrometry. The eight germacrane-type sesquiterpenoids showed nerve growth factor (NGF) potentiating activity, which mediates the neurite outgrowth in PC 12D cells. This study intends to reveal the chemical basis of the use of V. officinalis var. latiofolia as a dietary supplement.

Four new coumarinolignoids from seeds of Solanum indicum.

Activity-guided fractionation of seeds of Solanum indicum for anti-HBV activity led to the isolation of two novel coumarinolignoid alkaloids (indicumines A-B, 1-2) and two new coumarinolignoids (indicumines C-D, 3-4), together with four known coumarins (5-8). Their structures were established on the basis of spectroscopic data. The two novel coumarinolignoid alkaloids shows anti-HBV activities through specifically inhibiting the secretion of HBsAg in HepG2.2.15.

Two novel secoiridoid glucosides from Tripterospermum chinense.

Two novel secoiridoid glucosides, tripterospermumcins C (1) and D (2), were isolated from the aerial parts of Tripterospermum chinense, along with four known compounds, tripterospermumcin B (3), sweroside (4), loganic acid (5), and 8-epi-kingiside (6). Their structures were determined by analysis of 1D and 2D NMR data, as well as by comparison with model compounds. Compound 1 was a rare iridoid tetramer with four glucosides.

Sequence mining and transcript profiling to explore differentially expressed genes associated with lipid biosynthesis during soybean seed development.

BACKGROUND: Soybean (Glycine max L.) is one of the most important oil crops in the world. It is desirable to increase oil yields from soybean, and so this has been a major goal of oilseed engineering. However, it is still uncertain how many genes and which genes are involved in lipid biosynthesis. RESULTS: Here, we evaluated changes in gene expression over the course of seed development using Illumina (formerly Solexa) RNA-sequencing. Tissues at 15 days after flowering (DAF) served as the control, and a total of 11592, 16594, and 16255 differentially expressed unigenes were identified at 35, 55, and 65 DAF, respectively. Gene Ontology analyses detected 113 co-expressed unigenes associated with lipid biosynthesis. Of these, 15 showed significant changes in expression levels (log2fold values ≥ 1) during seed development. Pathway analysis revealed 24 co-expressed transcripts involved in lipid biosynthesis and fatty acid biosynthesis pathways. We selected 12 differentially expressed genes and analyzed their expressions using qRT-PCR. The results were consistent with those obtained from Solexa sequencing. CONCLUSION: These results provide a comprehensive molecular biology background for research on soybean seed development, particularly with respect to the process of oil accumulation. All of the genes identified in our research have significance for breeding soybeans with increased oil contents.

[Performance of new solid carbon source materials for denitrification].

Organic carbon is needed as the electron donor in the process of reduction of nitrate transformation to nitrogen gas, which is essential for biological denitrification. Based on previous research, agriculture wastes including corncob, rice hull, rice straw and sawdust were selected as potential carbon source for denitrification. Using the static organic material of carbon source leaching kinetics test and orthogonal experiments of external factors on carbon emission process, carbon release and its mechanism of a variety of carbon materials were studied. Study showed that release process of various types of carbon source materials follows the second dynamics formula, the release curve displayed a better double-reciprocal relationship. It revealed that release amount of rice straw was the highest and sawdust was the lowest. Results showed that corncob could better be used as carbon source for denitrification. Orthogonal test indicated that the increasing of solid-liquid ratio and water temperature would lead to an enhanced release capacity of carbon, however, the change of pH had no significant effect on release capacity of carbon; according to significant degree of water temperature, pH, solid-liquid ratio impacted on the carbon release, it was sorted by solid-liquid ratio > temperature > pH.

[Nitrate removal from wastewater using rice straw as carbon source and biofilm carrier].

In this research,agriculture waste-rice straw was investigated as the sole carbon source as well as biofilm carrier to remove nitrate from wastewater in up-flow laboratory reactors. The experimental results indicated that the startup of the system was quick and a high nitrate removal efficiency was observed. The accumulation of nitrite was not observed in this process. Moreover, high nitrate concentration in the influent had a significant effect on nitrate removal efficiency. The reactor was able to accommodate a wide range of pH (6.5-8.5) and DO (1.0-3.5 mg/L). A time-dependent decrease in nitrate removal efficiency was observed after 84 days of operation. The results showed that rice straw could be used as an economical and effective carbon source for denitrification.

Biological denitrification using corncobs as a carbon source and biofilm carrier.

In this research, agricultural waste--in particular, comcobs--was investigated for use as the sole carbon source and biofilm carrier to remove nitrate from wastewater in up-flow laboratory reactors. An artificial wastewater with a temperature range of 27 to 33 degrees C was used. Fast startup of the reactor and a high nitrate removal efficiency were observed. The highest denitrification rate of 0.203kg/(m3 x d) was achieved when flow rate and nitrate concentration were 153 L/d and 25.3 mgN/L, respectively. The accumulation of nitrite was not observed in this process. Moreover, flow rate and nitrate concentration of the influent were observed to have a significant effect on nitrate removal efficiency. A sharp decline of nitrate removal efficiency was observed when the flow rate was greater than 50 L/d. The reactor had the ability to accommodate a wide range of pH levels (6.5 to 8.5) and dissolved oxygen (1.5 mg/L to 4 mg/L). A time-dependent decrease in nitrate removal efficiency was observed after 67 days of operation. The addition of fresh corncobs brought about a rapid increase of nitrate removal efficiency. Results showed that corncobs could be used as an economical and effective carbon source for denitrification.

[Indirect determination of Vc with flame atomic absorption spectrometry].

In the present paper, a new method for indirect determination of Vc by atomic absorption spectrometry is proposed, based on the reducing properties of Ag+. The effects of temperature, reaction time and use level of Ag+ on the experiment were studied. Room-temperature and reaction time of 35 minutes were chosen. The oxidant amount is 2.0 mL solution of Ag+ (1.0 microg mL(-1)). Meanwhile the AAS working conditions for Ag determination were optimized. The proposed method allows the determination of Vc in a wide range with a relative standard deviation of 2.04%, and the detection limit is less than 1 microg x mL(-1). The method cannot be disturbed by the colour of sample. The interference of coexistent substance is also weak. Other methods for determining Vc could be remedied by this method. Two kinds of standard curves were plotted, standard working curve of sliver and standard working curve of Vc. The former is easier, while the latter is more accurate and could be applied flexibly according to the physical circumstances. This method is easy to control and has been applied to the determination of ascorbic acid in pharmaceutical preparations and orange juice. The recovery ratio of this method is 99.30%-106.06%. The results obtained in the analysis agreed well with the iodimetry in pharmacopeia.