ABSTRACT
Changes in the quality and quantity of litter and root inputs due to climate change and human activities can influence below-ground biogeochemical processes in forest ecosystems. However, it is unclear whether and how much aboveground litter and root inputs affect soil microbial metabolism and nutrient limitation mechanisms. In this study, according to a 4-years field manipulation experiment, litter and root manipulations (control (CK), double litter input (DL), no litter (NL), no root (NR), and no inputs (NI)) were set up to analyze the extracellular enzyme activities and stoichiometric ratios characteristics of 0-10 cm and 10-20 cm soils, explore the metabolic limitations of microorganisms, and clarify the main driving factors restricting nutrient limitation. The results showed that the enzyme activities associated with the C cycling (ß-1,4-glucosidase (BG), cellulose disaccharide hydrolase (CBH)) and N cycling (ß-1,4-N-acetylglucosaminidase (NAG), leucine aminopeptidase (LAP)) in DL treatment were significantly higher than those in NR treatment. Moreover, enzyme activities related to P cycling are significantly higher in comparison to other treatments. The acid phosphatase (AP), which is related to the P cycle, showed the highest activity under NR treatment. In addition, there was no significant difference in soil microbial metabolic limitation by the different carbon inputs, which did not change the original nutrient limitation pattern. The main drivers of microbial nutrient metabolic limitation included soil physicochemical properties, soil total nutrients, and available nutrients, among which soil SWC and pH presented the greatest influence on microbial C limitation and soil total nutrients showed the greatest influence on microbial N limitation. Changes in soil carbon input altered soil extracellular enzyme activities and their stoichiometric ratios by affecting soil physicochemical properties, total nutrients. This study provides data for the understanding of material cycling in forest ecosystems under environmental change.
Subject(s)
Ecosystem , Soil , Humans , Soil/chemistry , Carbon/metabolism , Soil Microbiology , Forests , Nutrients , Nitrogen/metabolism , Phosphorus/metabolismABSTRACT
BACKGROUND: Artemisia annua, a well-known traditional Chinese medicine, is the main source for production of artemisinin, an anti-malaria drug. A. annua is distributed globally, with great diversity of morphological characteristics and artemisinin contents. Diverse traits among A. annua populations impeded the stable production of artemisinin, which needs an efficient tool to identify strains and assess population genetic homogeneity. PURPOSE: In this study, ribosomal DNA (rDNA), were characterized for A. annua for strains identification and population genetic homogeneity assessment. METHODS: The ribosomal RNA (rRNA) genes were identified using cmscan and assembled using rDNA unit of LQ-9 as a reference. rDNA among Asteraceae species were compared performing with 45S rDNA. The rDNA copy number was calculated based on sequencing depth. The polymorphisms of rDNA sequences were identified with bam-readcount, and confirmed by Sanger sequencing and restriction enzyme experiment. The ITS2 amplicon sequencing was used to verify the stability of ITS2 haplotype analysis. RESULTS: Different from other Asteraceae species, 45S and 5S linked-type rDNA was only found in Artemisia genus. Rich polymorphisms of copy number and sequence of rDNA were identified in A. annua population. The haplotype composition of internal transcribed spacer 2 (ITS2) region which had moderate sequence polymorphism and relative short size was significantly different among A. annua strains. A population discrimination method was developed based on ITS2 haplotype analysis with high-throughput sequencing. CONCLUSION: This study provides comprehensive characteristics of rDNA and suggests that ITS2 haplotype analysis is ideal tool for A. annua strain identification and population genetic homogeneity assessment.
Subject(s)
Artemisia annua , Artemisinins , Asteraceae , Artemisia annua/genetics , DNA, Ribosomal/genetics , Medicine, Chinese TraditionalABSTRACT
The study of soil organic carbon components in continuous cropping cotton fields in oases is helpful to reveal the change characteristics of the soil organic carbon stability mechanism in arid areas under the effects of man-land relationships. In this study, the contents of soil organic carbon, easily oxidized organic carbon, dissolved organic carbon, and microbial biomass carbon in cotton fields with different continuous cropping years (2 a, 5 a, 12 a, 20 a, and 35 a) were collected and analyzed by using space instead of the time series method. Through redundancy analysis, the relationship between soil organic carbon components and other soil physical and chemical factors was discussed. The results showed that:â continuous cropping for different years had a significant impact on the content of soil organic carbon components in the study area. The contents of soil organic carbon, easily oxidized organic carbon, dissolved organic carbon, and microbial biomass carbon in continuous cropping cotton fields for 12 a, 20 a, and 35 a were higher than those in continuous cropping cotton fields and wasteland for 2 a and 5 a. ω(soil organic carbon) reached the peak value (7.06 g·kg-1) in the cotton field in 20 a, which was 76.91% higher than that in the wasteland. The content of soil organic carbon decreased with the deepening of the soil layer. â¡ Based on the redundancy analysis of soil organic carbon content and soil environmental factors, the results showed that the content of soil organic carbon was positively correlated with total nitrogen, available phosphorus, and water content and negatively correlated with pH value and bulk density. The importance of soil environmental factors on the interpretation of soil organic carbon content was as follows:total N>available P>pH value>bulk density>water content>available K>total salt.
Subject(s)
Carbon , Soil , Agriculture , Carbon/analysis , Humans , Nitrogen/analysis , Phosphorus/analysis , Soil/chemistry , Water/analysisABSTRACT
Soya saponin (SS) helps to improve antioxidant and immune function of body, and intestinal bacteria might play an important role here. In the present study, the co-occurring network of the ileal flora was analyzed with 50 mg/kg SS supplemented to the diet, and Romboutsia was found to have evolved into a dominant flora. In addition, the co-occurring network of the flora was changed with the combined antibiotic treated, and the unidentified-cyanobacteria developed into the dominant flora, whereas the relative abundance of Romboutsia was dropped. Dietary SS failed to elevate the relative abundance of Romboutsia with antibiotics treated, at the same time, it was not helpful for the antioxidant and immune function of laying hens. While dietary SS had a little help on the egg-laying performance. Intestinal bacteria did play a key role in the biological functions of SS on laying hens. In conclusion, SS failed to improve the antioxidation and immune function of laying hens with antibiotics treated.
Subject(s)
Antioxidants , Saponins , Animal Feed/analysis , Animal Nutritional Physiological Phenomena , Animals , Anti-Bacterial Agents/pharmacology , Chickens , Diet/veterinary , Dietary Supplements , Female , Immunity , Saponins/pharmacology , Glycine maxABSTRACT
BACKGROUND: Amomum villosum Lour. is the plant that produces the famous traditional Chinese medicine Amomi Fructus. Frequent habitat destruction seriously threatens A. villosum germplasm resources. Genetic diversity is very important to the optimization of germplasm resources and population protection, but the range of inherited traits within A. villosum is unclear. In this study, we analyzed the genetic diversity and genetic structures of A. villosum populations in Guangdong and constructed a local reference DNA barcode library as a resource for conservation efforts. METHODS: DNA barcoding and Inter-Simple Sequence Repeat (ISSR) markers were used to investigate the population genetics of A. villosum. Five universal DNA barcodes were amplified and used in the construction of a DNA barcode reference library. Parameters including percentage of polymorphic sites (PPB), number of alleles (Na), effective number of alleles (Ne), Nei's gene diversity index (H), and Shannon's polymorphism information index (I) were calculated for the assessment of genetic diversity. Genetic structure was revealed by measuring Nei's gene differentiation coefficient (Gst), total population genetic diversity (Ht), intra-group genetic diversity (Hs), and gene flow (Nm). Analysis of molecular variance (AMOVA), Mantel tests, unweighted pair-group method with arithmetic mean (UPGMA) dendrogram, and principal co-ordinates (PCoA) analysis were used to elucidate the genetic differentiation and relationship among populations. RESULTS: A total of 531 sequences were obtained from the five DNA barcodes with no variable sites from any of the barcode sequences. A total of 66 ISSR bands were generated from A. villosum populations using the selected six ISSR primers; 56 bands, 84.85% for all the seven A. villosum populations were polymorphic. The A. villosum populations showed high genetic diversity (H = 0.3281, I = 0.4895), whereas the gene flow was weak (Nm = 0.6143). Gst (0.4487) and AMOVA analysis indicated that there is obvious genetic differentiation amongA. villosum populations and more genetic variations existed within each population. The genetic relationship of each population was relatively close as the genetic distances were between 0.0844 and 0.3347.
ABSTRACT
This study aimed to explore the mechanism underlying arginine-promoted myogenesis of myoblasts. C2C12 cells were cultured with a medium containing 0.1, 0.4, 0.8, or 1.2 mmol/L arginine, respectively. Cell proliferation, viability, differentiation indexes, cytoplasmic Ca2+ concentration, and relative mRNA expression levels of myogenic regulatory factors (MRF) and key Ca2+ channels were measured in the absence or presence of 2 chemical inhibitors, dantrolene (DAN, 10 µmol/L) and nisoldipine (NIS, 10 µmol/L), respectively. Results demonstrated that arginine promoted myogenic differentiation and myotube formation. Compared with the control (0.4 mmol/L arginine), 1.2 mmol/L arginine upregulated the relative mRNA expression levels of myogenin (MyoG) and Myomaker at d 2 during myogenic induction (P < 0.05). Cytoplasmic Ca2+ concentrations were significantly elevated by arginine supplementation at d 2 and 4 (P < 0.05). Relative mRNA expression levels of Ca2+ channels including the type 1 ryanodine receptor (RyR1) and voltage-gated Ca2+ channel (Cav1.1) were upregulated by 1.2 mmol/L arginine during 2-d myogenic induction (P < 0.01). However, arginine-promoted myogenic potential of myoblasts was remarkably compromised by DAN and NIS, respectively (P < 0.05). These findings evidenced that the supplementation of arginine promoted myogenic differentiation and myotube formation through increasing cytoplasmic Ca2+ concentration from both extracellular and sarcoplasmic reticulum Ca2+.
ABSTRACT
The root of Reynoutria multiflora (Thunb.) Moldenke (syn: Polygonum multiflorum Thunb.) is a distinguished herb that has been popularly used in traditional Chinese medicine. The raw Reynoutria multiflora (RRM) should be processed by steaming before use, and the processing time is not specified in the processing specification. Our previous studies showed that the efficacy and toxicity of processed Reynoutria multiflora (PRM) at different processing times were inconsistent. A comprehensive identification method was established in this study to find a quality marker of raw Reynoutria multiflora (RRM) and processed Reynoutria multiflora (PRM) with different processing times. Metabolomics based on ultra-high-performance liquid chromatography tandem quadrupole/electrostatic field orbitrap high-resolution mass spectrometry (UHPLC-Q-Exactive plus orbitrap MS/MS) was used in this study. Using the CD.2 software processed database, multivariate statistical analysis methods coupled with cluster analysis and heatmap were implemented to distinguish between RRMs and PRMs with different processing times. The results showed that RRM and PRMs processed for 4, 8, 12, and 18 h cluster into group 1, and PRM processed for 24 and 32 h into group 2, indicating that it can effectively distinguish between the two groups and twenty potential markers, made the highest contributions to the observed chemical differences between two groups. Among them, tetrahydroxystilbene-O-hexoside-O-galloyl and sucrose can be used to identify PRM processed for 24 h. Therefore, the properties of RRM changed after 24 h of processing, and the quality markers were screened to distinguish RRM and PPM. It can also be used as an important control technology for the processing of RM, which has wide application prospects.
ABSTRACT
Exposures to toxic trace elements and deficiencies of essential elements during pregnancy are associated to various birth complications. Assessment of the trace elements in pregnant women living in specific areas is important for biomonitoring. A total of 196 healthy pregnant women absent of pregnancy complications living in Wuhan of China and 210 healthy non-pregnant women were enrolled. The whole blood were collected. The toxic element chromium (Cr), arsenic (As), cadmium (Cd), mercury (Hg), thallium (Tl), and lead (Pb) and essential elements magnesium (Mg), calcium (Ca), manganese (Mn), iron (Fe), copper (Cu), and zinc (Zn) were determined by using a inductively coupled plasma mass spectrometry (ICP-MS)-based method. All the metal(loid)s, except for Cd, Hg, and Tl, showed different levels in whole blood of the pregnant women compared with the non-pregnant women (p < 0.05), among which Mg, Fe, As, and Pb were lower while Ca, Cr, Mn, Cu, and Zn were higher. Moreover, whole blood levels of Mg, Mn, Fe, Cu, and Zn showed significant variations among different gestational ages, while As and Cd showed significant variations among different maternal ages. In addition, Fe-Mg, Fe-Zn, Cu-Ca, and Hg-As were found to be correlated positively in whole blood of the pregnant women, while Fe-Ca, Zn-Ca, and Fe-Cu were correlated negatively. The systematic information of toxic and essential elements in whole blood of pregnant women living in Wuhan of China can provide important guidance for the supplementation of essential elements during pregnancy and for biomonitoring of environmental overexposure.
Subject(s)
Pregnant Women , Trace Elements , Cadmium , China , Female , Humans , Pregnancy , Trace Elements/analysis , ZincABSTRACT
CRP (Citri Reticulatae Pericarpium), a famous traditional Chinese medicine, has also been extensively used in foods and condiments in dietary practice for centuries. According to the Chinese Pharmacopeia (2015 edition) it contains two subtypes, Guangchenpi (GCP) and Chenpi (CP). GCP exclusively originates from the pericarp of Citrus reticulata 'Chachi' cultivar and it's generally believed that GCP has superior qualities compared with the other main cultivars (CP). In the present study, an integrated approach combining LC-QTOF MS-based untargeted metabolomics analysis and DNA barcoding molecular identification was conducted to study the genetic diversity and chemical differences between GCP and CP. A validated UPLC-QTOF MS metabolomics method was established to identify markers by using PCA and OPLS-DA models. 34 identified metabolites could be used as chemical markers to distinguish effectively between the two subtypes. Among them polymethoxyflavones (PMF) such as hexamethoxyflavone (nobiletin and natsudaidain), pentamethoxyflavone (tangeretin and sinensetin), and tetramethoxyflavone are the most influential markers. Support vector machines were employed to classify all the samples and these markers showed good prediction accuracy (100%). The results of DNA barcoding showed that the secondary structure of the ITS2 sequences were significantly different among GCP and other three cultivars. The study indicated the integrated method could be a powerful and reliable analytical tool for differentiating GCP from CP.
ABSTRACT
Herb genomics and comparative genomics provide a global platform to explore the genetics and biology of herbs at the genome level. Panax ginseng C.A. Meyer is an important medicinal plant for a variety of bioactive chemical compounds of which the biosynthesis may involve transport of a wide range of substrates mediated by oligopeptide transporters (OPT). However, information about the OPT family in the plant kingdom is still limited. Only 17 and 18 OPT genes have been characterized for Oryza sativa and Arabidopsis thaliana, respectively. Additionally, few comprehensive studies incorporating the phylogeny, gene structure, paralogs evolution, expression profiling, and co-expression network between transcription factors and OPT genes have been reported for ginseng and other species. In the present study, we performed those analyses comprehensively with both online tools and standalone tools. As a result, we identified a total of 268 non-redundant OPT genes from 12 flowering plants of which 37 were from ginseng. These OPT genes were clustered into two distinct clades in which clade-specific motif compositions were considerably conservative. The distribution of OPT paralogs was indicative of segmental duplication and subsequent structural variation. Expression patterns based on two sources of RNA-Sequence datasets suggested that some OPT genes were expressed in both an organ-specific and tissue-specific manner and might be involved in the functional development of plants. Further co-expression analysis of OPT genes and transcription factors indicated 141 positive and 11 negative links, which shows potent regulators for OPT genes. Overall, the data obtained from our study contribute to a better understanding of the complexity of the OPT gene family in ginseng and other flowering plants. This genetic resource will help improve the interpretation on mechanisms of metabolism transportation and signal transduction during plant development for Panax ginseng.
Subject(s)
Ginsenosides/chemistry , Ginsenosides/metabolism , Magnoliopsida/metabolism , Panax/chemistry , Transcription Factors/metabolism , Gene Expression Regulation, Plant/genetics , Gene Expression Regulation, Plant/physiology , Genome, Plant/genetics , Magnoliopsida/genetics , Phylogeny , Transcription Factors/geneticsABSTRACT
Southern Chinese Medicine (SCM) is an important sect of Traditional Chinese Medicine (TCM) with its own special cultural style. Species identification is essential for TCM quality control because authentic herbs are possibly substituted with adulterants that would threaten the health of the public or even cause death. Here, we provided the first local reference DNA barcode library based on the second internal transcribed spacer (ITS2) for the molecular identification of SCM. A total of 1512 specimens of southern herbs representing 359 species were collected under the instructions and identification of taxonomic experts. Genomic DNA was extracted, and the PCR reaction proceeded according to standard procedures. After Sanger sequencing, sequence assembling and annotation, a reliable ITS2 barcode library with 1276 sequences from 309 species of Southern herbs was constructed. The PCR efficiency of the whole samples was 84.39%. Characteristics of the ITS2 barcode were analyzed, including sequence lengths and GC contents in different taxa. Neighbor-joining trees based on Kimura 2-Parameter (K2P) genetic distances showed a 67.56% successful rate of species identification with ITS2 barcode. In addition, 96.57% of species could be successfully identified at the genus level by the BLAST method. Eleven plant species were discovered to be cryptic. In addition, we found that there is an incorrect sequence existing in the public database, making a reliable local DNA barcode reference more meaningful. ITS2 barcodes exhibit advantages in TCM identification. This DNA barcode reference library could be used in Southern Chinese Medicine quality control, thus contributing to protecting public health.
Subject(s)
DNA Barcoding, Taxonomic/methods , DNA, Plant/genetics , Gene Library , Plants, Medicinal/genetics , China , Drugs, Chinese Herbal/metabolism , Genetic Variation , Medicine, Chinese Traditional , Quality ControlABSTRACT
The root of Polygonum multiflorum (PM) is an important Chinese herbal medicine for treatment of various diseases. Extensive pharmacological studies have been conducted and demonstrated that it shows a wide range of bioactivities. Meanwhile, a considerable number of hepatotoxicity cases owing to oral administration of PM have been reported and have attracted great attention. However, the limited knowledge regarding the metabolism of PM restricts the deeper studies on its pharmacological/toxicological mechanism and therapeutic material basis. The present study aimed to develop a high-performance liquid chromatography coupled with a linear ion trap-Orbitrap hybrid mass spectrometry method for separation and identification of metabolites in rat urine and plasma after oral administration of PM. Based on the proposed strategy, metabolism profiles of PM in rats were proposed for the first time and 43 metabolites were characterized or tentatively identified. Phase II metabolism, such as glucuronidation and sulfation, are the principal pathways of the main components. These findings will be beneficial to further understanding of the pharmacological mechanism and pharmacodynamic material basis of PM.
Subject(s)
Chromatography, High Pressure Liquid/methods , Drugs, Chinese Herbal/analysis , Drugs, Chinese Herbal/metabolism , Polygonum/chemistry , Spectrometry, Mass, Electrospray Ionization/methods , Animals , Drugs, Chinese Herbal/chemistry , Drugs, Chinese Herbal/pharmacokinetics , Emodin/analogs & derivatives , Male , Plant Roots/chemistry , Rats , Rats, Sprague-DawleyABSTRACT
We analyzed the differentiation among the environmental factors and soil organic/inorganic carbon contents of irrigated desert soil, brown desert soil, saline soil and aeolian sandy soil by classical statistics methods, and studied the correlation between soil carbon contents and the environmental factor by redundancy analysis (RDA) in a typical oasis of Yutian in the southern edge of the Tarim Basin. The results showed that the average contents of soil organic carbon and soil inorganic carbon were 2.51 g · kg⻹ and 25.63 g · kg⻹ respectively. The soil organic carbon content of the irrigated desert soil was significantly higher than those of brown desert soil, saline soil and aeolian sandy soil, while the inorganic carbon content of aeolian sandy soil was significantly higher than those of other soil types. The soil moisture and nutrient content were the highest in the irrigated desert soil and the lowest in the aeolian sandy sail. All soil types had high degree of salinization except the irrigated desert soil. The RDA results showed that the impacts of environmental factors on soil carbon contents ranked in order of importance were total nitrogen > available phosphorus > soil moisture > ground water depth > available potassium > pH > total salt. The soil carbon contents correlated extremely significantly with total nitrogen, available phosphorus, soil moisture and ground water depth (P < 0.01), and it correlated significantly with available potassium and pH (P < 0.05). There was no significant correlation between soil carbon contents and other environmental factors (P > 0.05).
Subject(s)
Carbon/analysis , Desert Climate , Soil/chemistry , Groundwater/analysis , Nitrogen/analysis , Phosphorus/analysis , Soil/classificationABSTRACT
OBJECTIVES: To compare the efficacy and safety of Tripterygium wilfordii Hook F (TwHF) with methotrexate (MTX) in the treatment of active rheumatoid arthritis (RA). METHODS: Design: a multicentre, open-label, randomised controlled trial. All patients were assessed by trained investigators who were unaware of the therapeutic regimen. INTERVENTION: 207 patients with active RA were randomly allocated (1:1:1) to treatment with MTX 12.5â mg once a week, or TwHF 20â mg three times a day, or the two in combination. At week 12, if reduction of the 28-joint count Disease Activity Score (DAS28) was <30% in the monotherapy groups, the patient was switched to MTX+TwHF. The primary efficacy point was the proportion of patients achieving an American College of Rheumatology (ACR) 50 response at week 24. RESULTS: 174/207 (84.1%) patients completed 24â weeks of the trial. In an intention-to-treat analysis, the proportion of patients reaching the ACR50 response criteria was 46.4% (32/69), 55.1% (38/69) and 76.8% (53/69), respectively, in the MTX, TwHF and MTX+TwHF groups (TwHF vs MTX monotherapy, p=0.014; MTX+TwHF vs MTX monotherapy, p<0.001). Similar statistically significant patterns at week 24 were found for ACR20, ACR70, clinical Disease Activity Index good responses, EULAR good response, remission rate and low disease activity rate. Significant improvement in the Health Assessment Questionnaire and 36-item Short-Form Health Survey questionnaire scores from baseline to week 24 was seen in each treatment arm (p<0.05), though no significant difference was found among the treatment arms (p>0.05). The result of per-protocol analysis agreed with that seen in the intention-to-treat analysis. Seven, three and five women in the TwHF, MTX and combination groups, respectively, developed irregular menstruation (TwHF vs MTX monotherapy, p=0.216). CONCLUSIONS: TwHF monotherapy was not inferior to, and MTX+TwHF was better than, MTX monotherapy in controlling disease activity in patients with active RA. TRIAL REGISTRATION NUMBER: NCT01613079.
Subject(s)
Antirheumatic Agents/therapeutic use , Arthritis, Rheumatoid/drug therapy , Methotrexate/therapeutic use , Phytotherapy , Plant Extracts/therapeutic use , Tripterygium , Adult , Female , Humans , Male , Middle Aged , Single-Blind Method , Treatment OutcomeABSTRACT
A new acidic polysaccharide (PLP) was isolated and characterized from Plantago asiatic L. seeds by hot alkali extraction and chromatographic purification using DEAE cellulose and Sephacryl S-400 columns. PLP has a molecular weight of 1.15 × 10(6) Da, and a monosaccharide composition of xylose (Xyl), arabinose (Ara), glucuronic acid (GlcA), and galactose (Gal) in a molar ratio of 18.8:7.2:6.1:1. The results of methylation analysis, FT-IR, and 1D and 2D NMR indicated that PLP was a highly branched heteroxylan of ß-1,4-linked Xylp backbone with three α-GlcAp-(1â3)-Araf attached to the O-3 position and one α-T-linked-GlcAp and one α-Araf-(1â5)-Araf attached to the O-2 position every eight monosaccharide residues. PLP exhibited scavenging abilities against hydroxyl, peroxyl anion, and DPPH radicals in vitro and showed significant binding capacities against cholic and chenodeoxycholic acids, suggesting its possible cholesterol-lowering activity. The results demonstrated the potential use of PLP in functional foods and nutraceuticals.
Subject(s)
Bile Acids and Salts/chemistry , Free Radical Scavengers/chemistry , Plant Extracts/chemistry , Plantago/chemistry , Polysaccharides/chemistry , Carbohydrate Sequence , Free Radical Scavengers/isolation & purification , Molecular Sequence Data , Plant Extracts/isolation & purification , Polysaccharides/isolation & purification , Seeds/chemistry , Spectroscopy, Fourier Transform InfraredABSTRACT
A novel polysaccharide (GPP-S), with a molecular mass of 1.2 × 10(6) Da, was isolated from the tetraploid Gynostemma pentaphyllum Makino by alkali extraction followed by purifications using DEAE and Sephacryl S-400 column chromatographies. The monosaccharide composition of GPP-S was determined as rhamnose, arabinose, glucose, and galactose with a molar ratio of 1.00:3.72:19.49:7.82. The structural analysis suggested that the backbone of GPP-S is (1â4)-linked-glucose and (1â6)-linked-galactose with a (1â4,6)-linked-glucose branch every six monosaccharide residues. The terminals were 1-)-α-arabinose, glucuronic acid, and other monosaccharides. GPP-S exhibited scavenging capacities against hydroxyl, peroxyl, and DPPH(â¢) radicals in vitro. GPP-S also had inhibitory activities on IL-1ß, IL-6, and COX-2 gene expressions in RAW 264.7 mouse macrophage cells. These results suggested that GPP-S could be developed as a bioactive ingredient for functional foods and dietary supplements.
Subject(s)
Anti-Inflammatory Agents/chemistry , Antioxidants/chemistry , Gynostemma/chemistry , Plant Extracts/chemistry , Polysaccharides/chemistry , Animals , Anti-Inflammatory Agents/pharmacology , Antioxidants/pharmacology , Gynostemma/genetics , Interleukin-1beta/immunology , Interleukin-6/immunology , Macrophages/drug effects , Macrophages/immunology , Mice , Molecular Structure , Plant Extracts/pharmacology , Polysaccharides/pharmacology , TetraploidyABSTRACT
Brucine, one of the main active ingredients in semen Strychni, has been included in many oral prescriptions of traditional Chinese medicine. In this study, we investigated the in vitro metabolism of brucine by human liver microsomes (HLMs) and the metabolic interactions of brucine with the substrates of cytochrome P450 (CYP450). Brucine was incubated with HLMs or CYP3A4 and then analysed by Liquid chromatography/mass spectrometry. The Km and Vmax values for HLMs were 30.53±3.14µM and 0.08±0.0029nmol/mg protein/min, respectively, while the corresponding values for CYP3A4 were 20.12±3.05µM and 6.40±0.21nmol/nmol P450/min. CYP3A4 may be the major enzyme responsible for brucine metabolism in HLMs, other human isoforms of CYP showed minimal or no effect on brucine metabolism. The inhibitory action of brucine was observed in CYP3A4 for the 1'-hydroxylation of midazolam, with inhibitory concentration 50 (IC50) of 8.4-fold higher than specific inhibitors in HLMs. Furthermore, brucine significantly inhibited the CYP3A4-catalyzed midazolam 1'-hydroxylation (Ki=2.14µM) at a concentration lower than 10µM, but no obvious inhibitory effects were observed on other CYP substrates (IC50>50µM). These results suggest that brucine has the potential to interact with a wide range of xenobiotics and endogenous chemicals especially CYP3A4 substrates.
Subject(s)
Cytochrome P-450 CYP3A/metabolism , Drugs, Chinese Herbal/metabolism , Microsomes, Liver/metabolism , Strychnine/analogs & derivatives , Biological Assay , Cells, Cultured , Drugs, Chinese Herbal/chemistry , Enzyme Activation/drug effects , Enzyme Inhibitors/pharmacokinetics , Humans , Hydroxylation/drug effects , Inhibitory Concentration 50 , Kinetics , Midazolam/metabolism , Strychnine/chemistry , Strychnine/pharmacokineticsABSTRACT
<p><b>OBJECTIVE</b>To evaluate the therapeutic effects and adverse effects of transarterial oily chemoembolization combined with interstitial laser thermotherapy (TOCE+ILT) in the treatment of hepatocellular carcinoma.</p><p><b>METHODS</b>Totally 120 patients with hepatocellular carcinoma were randomized into two groups and received interventions with TOCE+ILT or TOCE combined with percutaneous ethanol injection (TOCE+PEI). The treatment was repeated when necessary until the tumor was completely ablated, after which the therapeutic effects were evaluated and the patients were the followed up for observing long-term clinical outcome.</p><p><b>RESULTS</b>Of the 120 patients enrolled in this observation, 105 were followed up for two years (54 in TOCE+ILT group and 51 in TOCE+PEI group). The complete tumor necrosis rate of TOCE+ILT group was significantly higher than that of the TOCE+PEI group (84.8% vs 73.9%,Chi(2)=4.405, P=0.036), and TOCE+ILT was associated with a significantly higher negative conversion rate of AFP positivity (77.8% vs 56.1%, Chi(2)=4.592, P=0.032). The 1-year survival rate were similar between two groups, but the 2-year survival rate was significantly higher in patients with TOCE+ILT (79.6% vs 60.8%, Chi(2)=4.477, P=0.034). The hepatic function was comparable between the two groups before treatment, and 1 week after treatment, the ALT level in patients undergoing TOCE+ILT was significantly lower than that in patients with TOCE+PEI (95.90-/+56.06 U/L vs 116.31-/+45.27 U/L, t=2.04, P=0.043). Post-embolization syndrome was observed in the patients in two groups, but no severe adverse events were found.</p><p><b>CONCLUSION</b>TOCE+ILT has good therapeutic effects and mild side effects in the treatment of hepatocellular carcinoma.</p>