Danggui Sini decoction alleviates oxaliplatin-induced peripheral neuropathy by regulating gut microbiota and potentially relieving neuroinflammation related metabolic disorder.

BACKGROUND: Danggui Sini decoction (DSD), a traditional Chinese medicine formula, has the function of nourishing blood, warming meridians, and unblocking collaterals. Our clinical and animal studies had shown that DSD can effectively protect against oxaliplatin (OXA)-induced peripheral neuropathy (OIPN), but the detailed mechanisms remain uncertain. Multiple studies have confirmed that gut microbiota plays a crucial role in the development of OIPN. In this study, the potential mechanism of protective effect of DSD against OIPN by regulating gut microbiota was investigated. METHODS: The neuroprotective effects of DSD against OIPN were examined on a rat model of OIPN by determining mechanical allodynia, biological features of dorsal root ganglia (DRG) as well as proinflammatory indicators. Gut microbiota dysbiosis was characterized using 16S rDNA gene sequencing and metabolism disorders were evaluated using untargeted and targeted metabolomics. Moreover the gut microbiota mediated mechanisms were validated by antibiotic intervention and fecal microbiota transplantation. RESULTS: DSD treatment significantly alleviated OIPN symptoms by relieving mechanical allodynia, preserving DRG integrity and reducing proinflammatory indicators lipopolysaccharide (LPS), IL-6 and TNF-α. Besides, DSD restored OXA induced intestinal barrier disruption, gut microbiota dysbiosis as well as systemic metabolic disorders. Correlation analysis revealed that DSD increased bacterial genera such as Faecalibaculum, Allobaculum, Dubosiella and Rhodospirillales_unclassified were closely associated with neuroinflammation related metabolites, including positively with short-chain fatty acids (SCFAs) and sphingomyelin (d18:1/16:0), and negatively with pi-methylimidazoleacetic acid, L-glutamine and homovanillic acid. Meanwhile, antibiotic intervention apparently relieved OIPN symptoms. Furthermore, fecal microbiota transplantation further confirmed the mediated effects of gut microbiota. CONCLUSION: DSD alleviates OIPN by regulating gut microbiota and potentially relieving neuroinflammation related metabolic disorder.

Metals/bisulfite system involved generation of 24-sulfonic-25-ene ginsenoside Rg1, a potential quality control marker for sulfur-fumigated ginseng.

Ginseng is a most popular health-promoting food with ginsenosides as its main bioactive ingredients. Illegal sulfur-fumigation causes ginsenosides convert to toxic sulfur-containing derivatives, and reduced the efficacy/safety of ginseng. 24-sulfo-25-ene ginsenoside Rg1 (25-ene SRg1), one of the sulfur-containing derivatives, is a potential quality control marker of fumigated ginseng, but with low accessibility owing to its unknown generation mechanism. In this study, metals/bisulfite system involved generation mechanism was investigated and verified. The generation of 25-ene SRg1 in sulfur-fumigated ginseng is that SO2, formed during sulfur-fumigation, reacted with water and ionized into HSO3-. On the one hand, under the metals/bisulfite system, HSO3- generates HSO5- and free radicals which converted ginsenoside Rg1 to 24,25-epoxide Rg1; on the other hand, as a nucleophilic group, HSO3- reacted with 24,25-epoxide Rg1 and further dehydrated to 25-ene SRg1. This study provided a technical support for the promotion of 25-ene SRg1 as the characteristic quality control marker of sulfur-fumigated ginseng.

Rationality of the ethanol precipitation process in modern preparation production of Zishui-Qinggan decoction evaluated by integrating UPLC-QTOF-MS/MS-based chemical profiling/serum pharmacochemistry and network pharmacology.

INTRODUCTION: Zishui-Qinggan decoction (ZQD) is a classical traditional Chinese medicine formula (TCMF) for alleviating menopausal symptoms (MPS) induced by endocrine therapy in breast cancer patients. In the production of TCMF modern preparations, ethanol precipitation (EP) is a commonly but not fully verified refining process. OBJECTIVES: Chemical profiling/serum pharmacochemistry and network pharmacology approaches were integrated for exploring the rationality of the EP process in the production of ZQD modern preparations. MATERIAL AND METHODS: Ultra-performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (UPLC-QTOF-MS/MS) was applied to identify the chemical profiles and absorbed components of ZQD. Network pharmacology was used to identify targets and pathways related to MPS-relieving efficacy. RESULTS: The chemicals of ZQDs without/with EP process (referred to as ZQD-W and ZQD-W-P, respectively) were qualitatively similar with 89 and 87 components identified, respectively, but their relative contents were different; 51 components were detectable in the serum of rats orally administered with ZQD-W, whereas only 19 were detected in that administered with ZQD-W-P. Key targets, such as AKT1, and pathways, such as the PI3K-Akt signalling pathway, affected by ZQD-W and ZQD-W-P were similar, while the neuroactive ligand-receptor interaction pathway among others and the MAPK signalling pathway among others were specific pathways affected by ZQD-W and ZQD-W-P, respectively. The specifically absorbed components of ZQD-W could combine its specific key targets. CONCLUSION: The EP process quantitatively altered the chemical profiles of ZQD, subsequently affected the absorbed components of ZQD, and then affected the key targets and pathways of ZQD for relieving MPS. The EP process might result in variation of the MPS-relieving efficacy of ZQD, which deserves further in vivo verification.

Lyophilization enhances the stability of Panax notoginseng total saponins-loaded transfersomes without adverse effects on ex vivo/in vivo skin permeation.

Transfersomes (TFSs) have been extensively investigated to enhance transdermal drug delivery. As a colloidal dispersion system, TFSs are prone to problems such as particle aggregation and sedimentation, oxidation and decomposition of phospholipids. To enhance the stability of panax notoginseng saponins (PNS)-loaded transfersomes (PNS-TFSs) without adverse influences on their skin permeation, we prepared lyophilized PNS-loaded transfersomes (PNS-FD-TFSs), clarified their physicochemical characteristics and investigated their in vitro drug release, ex vivo skin permeation/deposition and in vivo pharmacokinetics. In this study, a simple, fast and controllable process was developed for preparing lyophilized PNS-TFSs. In the optimized PNS-FD-TFS formulation, sucrose and trehalose were added to the PNS-TFS dispersion with a mass ratio of trehalose, sucrose, and phospholipid of 3:2:1, and the mixture was frozen at -80 °C for 12 h followed by lyophilization at -45 °C and 5 Pa for 24 h. The optimized formulation of PNS-FD-TFSs was screened based on the appearance and reconstitution time of the lyophilized products, vesicle size, and PDI of the freshly reconstituted dispersions. It maintained stable physicochemical properties for at least 6 months at 4 °C. The vesicle size of PNS-FD-TFSs was below 100 nm and homogenous with a polydispersity index of 0.2 after reconstitution. The average encapsulation efficiencies of the five index saponins notoginsenoside R1 (NGR1), ginsenoside Rg1 (GRg1), ginsenoside Re (GRe), ginsenoside Rb1 (GRb1) and ginsenoside Rd (GRd) in PNS-FD-TFSs were 68.41 ± 5.77%, 68.95 ± 6.08%, 65.46 ± 10.95%, 91.50 ± 5.62% and 95.78 ± 1.70%, respectively. The reconstituted dispersions of PNS-FD-TFSs were similar to PNS-TFSs in in vitro release, ex vivo skin permeation, and deposition. The pharmacokinetic studies showed that, compared with the PNS liposomes (PNS-LPS), the PNS-FD-TFS-loaded drug could permeate through the skin and enter the blood rapidly. It can be concluded that the lyophilization process can effectively improve the stability of PNS-TFSs without compromising their transdermal absorption properties.

Unraveling the evolutionary dynamics of the TPS gene family in land plants.

Terpenes and terpenoids are key natural compounds for plant defense, development, and composition of plant oil. The synthesis and accumulation of a myriad of volatile terpenoid compounds in these plants may dramatically alter the quality and flavor of the oils, which provide great commercial utilization value for oil-producing plants. Terpene synthases (TPSs) are important enzymes responsible for terpenic diversity. Investigating the differentiation of the TPS gene family could provide valuable theoretical support for the genetic improvement of oil-producing plants. While the origin and function of TPS genes have been extensively studied, the exact origin of the initial gene fusion event - it occurred in plants or microbes - remains uncertain. Furthermore, a comprehensive exploration of the TPS gene differentiation is still pending. Here, phylogenetic analysis revealed that the fusion of the TPS gene likely occurred in the ancestor of land plants, following the acquisition of individual C- and N- terminal domains. Potential mutual transfer of TPS genes was observed among microbes and plants. Gene synteny analysis disclosed a differential divergence pattern between TPS-c and TPS-e/f subfamilies involved in primary metabolism and those (TPS-a/b/d/g/h subfamilies) crucial for secondary metabolites. Biosynthetic gene clusters (BGCs) analysis suggested a correlation between lineage divergence and potential natural selection in structuring terpene diversities. This study provides fresh perspectives on the origin and evolution of the TPS gene family.

Real-time online monitoring technology for sweeping frequency ultrasound (SFU) assisted extraction of amur grape (Vitis amurensis) seed oil.

Sweeping frequency ultrasound (SFU) was used to assist extraction of amur grape (Vitis amurensis) seed (AGS) oil. Extraction conditions and physicochemical properties were optimized and analyzed under different extraction methods. Meanwhile, frequency and time domains were online monitored during SFU assisted extraction of AGS oil. PVDF piezoelectric sensor was used in time domain, and the hydrophone in frequency domain, so as to obtain the time-voltage waveform, signal power, spectrum distribution and other visual models. Physical models of the spatial peak acoustic intensity, charge quantity and work done by electric field force under different ultrasonic conditions were derived. The mathematical model between the work done by electric field force and the spatial peak acoustic intensity under the working state of PVDF piezoelectric sensor was constructed. Results show that the content of AGS oil by SFU assisted extraction was higher than that by organic extraction. Furthermore, the optimal single-frequency was 40 kHz and dual-frequency was 28/33 kHz, and SFU extraction time of 30 min was suitable with higher oil yield of 16.70 % and 16.94 %, respectively. In addition, the selection and combination of SFU also affected the oil oxidation degree. The peak voltage, spatial peak acoustic intensity, signal power and work of electric field force at 28/33 kHz were all higher than those at 40 kHz.

Study of okra pectin prepared by sweeping frequency ultrasound/freeze-thaw pretreatment on corrosion inhibition of ANSI 304 stainless steel in acidic environment.

Green and efficient metal corrosion inhibitors are very essential, and natural okra pectin (OP) can fulfill this need with rational use of resources. OP was prepared by water-alcohol extraction method after freeze-thaw pretreatment (FTP)/sweeping frequency ultrasound pretreatment (SFUP), and used for corrosion inhibition of ANSI 304 stainless steel (304 SS) in 1 M hydrochloric acid (HCl). The molecular weight, hydrodynamic diameter and monosaccharide composition of OP were analyzed to determine the factors on the corrosion inhibition of 304 SS. During SFUP of okra, the time-domain variation of ultrasound field was monitored by piezoelectric film sensor, its frequency-domain variation was monitored by a hydrophone, and analyzed respectively by oscilloscope and spectrum analyzer. Static weight-loss method, electrochemical and microscopic analyses were used to evaluate the corrosion inhibition efficiency of OP at temperatures (25, 30, 40, 50 °C) and concentrations (0, 0.2, 0.5, 1, 2 g·L-1) to optimize corrosion inhibition performance. It was found that OP by FTP and SFUP had higher corrosion inhibition efficiency on metals in acidic environment. According to static weight-loss method, the corrosion inhibition efficiency of OP with concentration of 2 g·L-1 (25 °C) was improved to 90.27 % in the FTP group and 93.53 % in the SFUP group, which 5.14 % and 8.93 % higher than Control (without pretreatment). Meanwhile, the corrosion inhibition efficiency decreased gradually as the temperature increased. OP corrosion inhibition performance fit Langmuir adsorption isothermal model as a mixed adsorption based on physical adsorption. It was a mixed inhibitor to protect 304 SS from corrosion.

Mechanisms of Rostellularia procumbens (L.) Nees on treating chronic glomerulonephritis explored by network pharmacology, RNA-seq, and in vitro experiments.

BACKGROUND: The purpose of this study was to demonstrate the in vitro anti-nephritis activity of Rostellularia procumbens (L.) Nees (R. procumbens) extract and to make a preliminary investigation of its anti-nephritis mechanism. METHODS: A prediction network was built that describes the relationship between R. procumbens and CGN. Then, the potential targets for R. procumbens against CGN were imported into the DAVID database for Gene Ontology (GO) biological annotation analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. A lipopolysaccharide (LPS)-stimulated rat mesangial cell HBZY-1 model in vitro was used to examine the anti-inflammatory activity of R. procumbens extract. RNA-seq was utilized to investigate differentially expressed genes (DEGs) and enriched signaling pathways between groups. Finally, qPCR was used for the validation analysis of the experimental results. RESULTS: The results of network pharmacology showed that R. procumbens exerts its therapeutic effect on CGN through the AGE-RAGE signaling pathway in diabetic complications, PI3K-Akt, IL-17 signaling pathway, and so on. R. procumbens n-butanol extract (J-NE) can effectively relieve inflammation in HBZY-1. The results of KEGG pathway enrichment suggest that J-NE attenuated CGN was associated with the IL-17 signaling pathway, and the results of RNA-seq were consistent with network pharmacology. Targets enriched in the IL-17 signaling pathway, including Chemokine (C-C motif) ligand 7 (CCL7), Lipocalin 2 (LCN2), Chemokine (C-C motif) ligand 2 (CCL2), and Chemokine (C-X-C motif) ligand 1 (CXCL1), have been identified as crucial targets attenuating CGN by J-NE. CONCLUSION: R. procumbens is a promising pharmacological candidate for the treatment of CGN in the present era.

Dietary docosahexaenoic acid plays an opposed role in ferroptotic and non-ferroptotic acute kidney injury.

Ferroptosis due to polyunsaturated fatty acid (PUFA) peroxidation has been implicated in the pathogenesis of acute kidney injury (AKI), suggesting the risk of dietary intake of PUFA for people susceptible to AKI. Clinically, however, in addition to ferroptosis, other mechanisms also contribute to different types of AKI such as inflammation associated necroptosis and pyroptosis. Therefore, the role of PUFA, especially ω3 PUFA which is a common food supplement, in various AKIs deserves further evaluation. In this study, rhabdomyolysis- and folic acid-induced AKI (Rha-AKI and FA-AKI) were established in mice fed with different fatty acids Histology of kidney, blood urea nitrogen and creatinine, lipid peroxidation, and inflammatory factors were examined. Results showed that these two types of AKIs had diametrically different pathogenesis indicated by that ferrostatin-1 (Fer-1), a lipid antioxidant, can attenuate FA-AKI rather than Rha-AKI. Further, dietary DHA (provided by fish oil) reduced tubular injury and renal lesion by inhibiting peroxidation and inflammation in mice with Rha-AKI while increasing cell death, tissue damage, peroxidation and inflammation in mice with FA-AKI. In human renal tubular epithelial cell line HK-2, MTT assay and DHE staining showed that both myoglobin and ferroptosis inducers can cause cell death and oxidative stress. Ferroptosis inducer-induced cell death was promoted by DHA, while such result was not observed in myoglobin-induced cell death when adding DHA. This study illustrates that the mechanisms of AKI might be either ferroptosis dependent or -independent and the deterioration effect of dietary DHA depends on whether ferroptosis is involved.

UPLC-QTOF-MS/MS assisted UPLC-TQ-MS/MS strategy to comparatively investigate the rat pharmacokinetics of N-acetyldopamine oligomers derived from Cicadae Periostracum.

Cicadae Periostracum (CP), the slough molted from the nymph of Cryptotympana pustulata, is a widely used medicinal material in traditional Chinese medicine (TCM). N-acetyldopamine oligomers (NAOs), the homologues of acetyldopamine, including N-acetyldopamine dimers/trimers/tetramers/pentamers (NADs/NATrs/NATes/NAPs), side-chain isomer of dimers/trimers (SCIDs/SCITrs), are major bioactive ingredients of CP. However, owing to commercially unavailable reference substances of all NAOs, simultaneous quantification of these NAOs in biological samples is difficult, and thus their pharmacokinetics are still unknown. In this study, a comprehensive strategy for simultaneous quantification/semi-quantification of NAOs in plasma with single N-acetyldopamine dimer A (NAD-A) as reference substance was established and comparatively investigated their pharmacokinetics after oral administration of pure NAD-A and two types of CP extracts, i.e., post-molting-washed slough (CP-WAT) and pre-molting-washed slough (CP-WBT). A UPLC-QTOF-MS/MS assisted UPLC-TQ-MS/MS strategy was developed to quantify NAOs in rat plasma. NAOs in CP extract were qualitatively characterized by UPLC-QTOF-MS/MS, then the quasi-molecular ions and characteristic fragment ions of the identified NAOs by UPLC-QTOF-MS/MS were transferred to UPLC-TQ-MS/MS as parent-daughter ion pairs for MRM mode quantification of the NAOs, and the method was validated with single NAD-A for quantifying NAD-A and semi-quantifying other NAOs in plasma. The established method was applied to compare the pharmacokinetics of NAOs after oral administration of NAD-A and the extracts of CP-WBT/CP-WAT respectively. Six quasi-molecular ions and characteristic fragment ion m/z 192.1 were characterized by UPLC-QTOF-MS/MS and transferred to be the parent-daughter ion pairs for UPLC-TQ-MS/MS analysis of six kinds of NAOs. For the pharmacokinetics, NAD-A showed double peaks absorption character when administered with single compound, but with higher relative bioavailability when administered with CP extracts with the similar dosage. Compared with CP-WAT, NAOs in CP-WBT reached the maximum plasma concentration in much shorter time.

Comparing steamed and wine-stewed Rehmanniae Radix in terms of Yin-nourishing effects via metabolomics and microbiome analysis.

ETHNOPHARMACOLOGICAL RELEVANCE: Rehmanniae Radix Praeparata (RRP), the processed root of Rehmannia glutinosa, has been widely used to treat Yin deficiency syndrome in traditional Chinese medicine. RRP is available in two forms: processed by steaming with water (SRR) or processed by stewing with yellow rice wine (WRR). Previous work has documented chemical differences in the secondary metabolomes and glycomes of SRR and WRR. AIM OF THE STUDY: This study aimed to compare SRR and WRR in terms of Yin-nourishing effects via metabolomics and microbiome analysis. MATERIALS AND METHODS: ICR mice were orally administered with thyroxine for 14 d to induce Yin deficiency. Changes in biochemical indices and histopathology were detected. Serum metabolomics analysis and microbial 16S rRNA sequencing were performed to compare the therapeutic effects and mechanisms between SRR and WRR in treating thyroxine-induced Yin deficiency. RESULTS: Both SRR and WRR decreased serum T3, T4 and MDA levels, and increased SOD activity. SRR more effectively decreased serum Cr, and ameliorated kidney injury, while WRR showed better regulation on ratio of cAMP/cGMP and serum TSH, and relieved thyroid injury. Both SRR and WRR regulated tyrosine, glycerophospholipid, and linoleic acid metabolism and the citric acid cycle. Additionally, SRR regulated fatty acid metabolism, while WRR influenced alanine, aspartate and glutamate metabolism, and bile acid biosynthesis. SRR significantly enriched the genera Staphylococcus and Bifidobacterium in the gut microbiome, while WRR significantly enriched the genera Akkermansia, Bacteroides and Parabacteroides, and decreased the abundance of Lactobacillus. CONCLUSIONS: SRR displayed better protective effects on kidney, while WRR showed stronger effects on thyroid in thyroxine-induced Yin deficient mice. These differences might be due to different regulating effects of SRR and WRR on the metabolome and gut microbiota.

Holistic quality evaluation of Callicarpae Formosanae Folium by multi-chromatography-based qualitative and quantitative analysis of polysaccharides and small molecules.

Callicarpae Formosanae Folium (CFF), derived from the leaves of Callicarpa formosana Rolfe, is a common Chinese medicinal herb used for the treatment of hematemesis. Phytochemical studies found that phenylpropanoids, flavonoids, terpenoids and polysaccharides were the main ingredients of CFF. However, there is limited scientific information concerning holistic quality method and quality consistency evaluation of CFF. In this study, a strategy integrating HPGPC-ELSD, HPLC-PDA, UV-VIS and UPLC-QTOF-MS/MS was firstly developed to simultaneously qualify and quantify polysaccharides, as well as representative small molecules in CFF. HPGPC-ELSD was applied to characterize the molecular weight distribution of polysaccharides, HPLC-PDA was developed to qualitatively and quantitatively determine monosaccharides. UV-VIS was used to determine the total polysaccharides content, and UPLC-QTOF-MS/MS was established to characterize the small molecules. The quality consistency of commercial CFF (CM-CFF) was also evaluated. It was shown that the relative molecular weights, the compositional monosaccharides and small molecules composition in CM-CFF and self-collected CFF (SC-CFF) samples were similar. A total of 32 small molecules including 6 phenylpropanoids, 7 flavonoids and 19 terpenoids were characterized in CFF. However, the variation was observed in the content of polysaccharides, luteolin, ursolic acid, as well as total contents of terponoids in CM-CFF samples, which implied that the holistic quality of CM-CFF was inconsistent. The results suggested that the proposed evaluation strategy could be applied as a potential approach for the quality control of CFF. And the quality of CM-CFF should be improved by Good Agriculture Practice (GAP) base and standard processing method.

Impact of sulfur-fumigation on carbohydrate components of Atractylodis Macrocephalae Rhizoma.

Atractylodis Macrocephalae Rhizoma (AMR) is one of commonly used medicinal and edible herbs in China. It is often sulfur-fumigated during post-harvest processing. Carbohydrates are important active components of AMR. However, it is unknown whether sulfur-fumigation would induce changes on carbohydrates. Here, carbohydrates including polysaccharides, oligosaccharides and free monosaccharides were comprehensively analyzed to characterize the quality changes of sulfur-fumigated AMR. Determination of both homemade sulfur-fumigated AMR samples and commercial samples from market revealed that sulfur-fumigation did not affect molecular weight distribution of polysaccharides, but altered polysaccharides content and its ratios of constituent monosaccharides, especially glucose (Glc) and fructose (Fru), as well as the contents of oligosaccharides DP2-10 and free monosaccharide Fru. Moreover, the variations enhanced with the increasing of residual SO2 content. The potential transformation mechanisms could be due to the hydrolysis of polysaccharides. The research outcomes could provide a chemical basis for the safety and efficacy evaluations of sulfur-fumigated AMR.

Integrating serum pharmacochemistry and network pharmacology to identify chemical markers for quality control of Apocyni Veneti Folium.

INTRODUCTION: Apocyni Veneti Folium (AVF) is a commonly used traditional Chinese medicinal herb for the treatment of hypertension. Chemical markers are crucial for the quality control of herbal medicines; however, the therapeutic components of AVF remain to be well elucidated. OBJECTIVES: This study was intended to integrate serum pharmacochemistry and network pharmacology to identify chemical markers of AVF and establish an efficacy-related quality control method of AVF. MATERIAL AND METHODS: Ultra-performance liquid chromatography quadrupole time-of-flight tandem mass spectrometry (UPLC-QTOF-MS/MS) was applied to identify the absorbed AVF constituents in rat serum. Network pharmacology was further used to identify anti-hypertension-related chemical markers. Subsequently, a quantitative method was established using UPLC with diode array detection (DAD) and applied for quality evaluation of commercial AVF samples. RESULTS: Thirteen prototype constituents were unequivocally or tentatively characterized in serum samples, among which quercetin, kaempferol, hyperoside, isoquercitrin, chlorogenic acid, cryptochlorogenic acid, and neochlorogenic acid were identified as dominant chemicals related to anti-hypertensive efficacy. The quantitative data showed that the total contents of seven marker components even showed 2-fold variation among 14 batches of commercial AVF samples with RSD values ranging from 12.15% to 75.61%. Hierarchical cluster analysis and heatmap analysis showed that 14 batches of commercial AVF samples could be divided into three main groups. CONCLUSION: The chemical markers obtained from this study could be applicable for efficacy-related quality control of AVF.

Quality consistency evaluation of commercial Prunellae Spica by integrating determination of secondary metabolites and saccharides.

INTRODUCTION: Prunellae Spica (PS) is a commonly used medicinal herb in China. Secondary metabolites and saccharides are major bioactive components of PS. However, holistic quality consistency of commercial PS is ambiguous due to lack of comprehensive evaluation methods and reliable quality control markers. OBJECTIVES: Integrating multiple chromatographic and chemometric methods to comprehensively evaluate the holistic quality of PS. MATERIAL AND METHODS: Ultrahigh-performance liquid chromatography coupled with triple-quadrupole tandem mass spectrometry (UPLC-TQ-MS/MS) was applied to quantify 12 secondary metabolites of PS. High-performance liquid chromatography coupled with photodiode array/evaporative light scattering detection (HPLC-PDA/ELSD) and high-performance gel permeation chromatography (HPGPC) methods were used to characterise the saccharides. Multivariate statistical analysis was adopted to evaluate the quality consistency of commercial PS and explore the potential quality control markers. RESULTS: The contents of secondary metabolites and saccharides were significantly different among commercial PS. All samples could be classified into three groups with ferulic acid, protocatechualdehyde, gallic acid, ursolic acid/oleanolic acid, sucrose, p-coumaric acid, chlorogenic acid as the major contributing components responsible for the difference. The content of rosmarinic acid was correlated with that of betulinic acid, hyperposide, chlorogenic acid, rutin, caffeic acid, p-coumaric acid and glucose, whereas polysaccharides, ferulic acid, protocatechualdehyde and ursolic acid/oleanolic acid, quercetin, sucrose and majority monosaccharides were not. CONCLUSION: The holistic quality of commercial PS was inconsistent. Together with rosmarinic acid, ferulic acid, protocatechualdehyde, ursolic acid/oleanolic acid, polysaccharides and sucrose might be recommended as potential quality control markers for the holistic quality control of PS.

Supplementing Diets with Agriophyllum squarrosum Reduced Blood Lipids, Enhanced Immunity and Anti-Inflammatory Capacities, and Mediated Lipid Metabolism in Tan Lambs.

Agriophyllum squarrosum (sand rice), a widespread desert plant, possesses anti-hyperglycemic and anti-inflammatory properties, and has been used in traditional Chinese medicine for many years. However, its effects on ruminants are unknown. To fill this gap, we examined the effects of A. squarrosum on the immune and anti-inflammatory responses of lambs. A total of 23, 6-month-old Tan ewe-lambs (27.6 ± 0.47 kg) were divided into four groups and offered a basic diet (C­control), or a diet that contained 10%, 20%, or 30% A. squarrosum, on a dry matter basis, for 128 days. Serum concentrations of total cholesterol were lower (p = 0.004) in the 30% supplemented lambs than controls, while concentrations of high-density lipoprotein cholesterol were lower (p = 0.006) in the 10% and 20%, but not in 30% supplemented lambs than controls. Serum-cortisol concentrations were lower (p = 0.012) in the 30% supplemented lambs and free fatty acid concentrations were higher in the 10% and 20% supplemented lambs than in control lambs (p < 0.001). Supplementation with A. squarrosum decreased (p < 0.05) the area of adipocytes in subcutaneous adipose tissue, but there was no difference between the 20% and 30% diets. Conversely, the area in visceral adipose tissue (VAT) increased (p < 0.05), especially for the 10% and 20% supplemented diets. Supplementation with A. squarrosum also enriched immune and anti-inflammatory related and lipid and glucose-metabolic pathways and associated differentially expressed gene expressions in adipose tissue. A total of 10 differential triacylglycerol, 34 differential phosphatidylcholines and seven differential phosphatidylethanolamines decreased in the diet with 30% supplementation, when compared to the other diets. Finally, adipocyte-differentiation genes, and immune and inflammatory response-related gene expression levels decreased in lamb adipocytes cultured with an aqueous A. squarrosum extract. In conclusion, supplementing lamb diets with A. squarrosum reduced blood lipids, enhanced immunity and anti-inflammatory capacities, and mediated lipid metabolism in adipose tissue and adipocytes of Tan lambs. A level of approximately 10% is recommended, but further research is required to determine the precise optimal level.

Integrating Multi-Type Component Determination and Anti-Oxidant/-Inflammatory Assay to Evaluate the Impact of Pre-Molting Washing on the Quality and Bioactivity of Cicadae Periostracum.

Cicadae Periostracum (CP) is a traditional Chinese medicinal herb derived from the slough that is molted from the nymph of the insect Cryptotympana pustulata Fabricius. Washing with water to remove residual silt is a primary processing method of CP that is recommended by the Chinese Pharmacopoeia, but how washing methods affect the quality and bioactivity of CP is unknown. In this study, the quality and bioactivity of non-washed CP (CP-NW), post-molting-washed CP (CP-WAT), and pre-molting-washed CP (CP-WBT) were comparatively investigated. The quality of these CP samples was evaluated in terms of the UPLC-QTOF-MS/MS-based chemical profiling and semi-quantification of 39 N-acetyldopamine oligomers (belonging to six chemical types), the HPLC-UV-based quantification of 17 amino acids, the ICP-MS-based quantification of four heavy metals, and the contents of ash; the bioactivities of the samples were compared regarding their anti-oxidant and anti-inflammatory activities. It was found that, compared with CP-NW, both CP-WBT and CP-WAT had significantly lower contents of ash and heavy metals. Moreover, compared with CP-WAT, CP-WBT contained lower levels of total ash, acid-insoluble ash, and heavy metals and higher contents of N-acetyldopamine oligomers and amino acids. It also had enhanced anti-oxidant and anti-inflammatory activities. A Spearman's correlation analysis found that the contents of N-acetyldopamine oligomers and free amino acids were positively correlated with the anti-oxidant/-inflammatory activities of CP. All these results suggest that pre-molting washing can not only remove the residual silt but can also avoid the loss of the bioactive components and assure higher bioactivities. It is concluded that pre-molting washing could enhance the quality and bioactivity of CP and should be a superior alternative method for the primary processing of qualified CP.

Distribution Characteristics and Source Analysis of Nitrogen and Phosphorus in Different Rivers in Two Water Period: A Case Study of Pi River and Shiting River in the Upper Reaches of Tuo River in China.

In this paper, the distribution characteristics of total nitrogen (TN), total phosphorus (TP) and fractions of nitrogen and phosphorus in water and surface sediments of the Pi and Shiting rivers in the dry and wet seasons were studied by molybdenum blue/ascorbic acid spectrophotometry and Standard Measurements and Testing (SMT). Correlation analysis, cluster analysis and principal component analysis were used to identified nitrogen and phosphorus pollution sources. The results showed that: (1) nitrogen and phosphorus in water and surface sediments in the study area were at different levels. (2) In the Pi river, the decomposition of animal and plant residues, the leachate from the accumulation of aquaculture wastewater and urban domestic sewage were the main sources of nitrogen and phosphorus pollution, while in the Shiting river, the unreasonable application of pesticides and fertilizers, the degradation of animal and plant residues, agricultural wastewater from agricultural drainage channels, industrial production wastewater and the weathering of agricultural wastes had a great impact on the nitrogen and phosphorus pollution. The results in this study provide reliable experimental data and a reference to local relevant departments for the implementation of effective control measures for the reduction of the nitrogen and phosphorus pollution load in the river basin.

Co-existing polysaccharides affect the systemic exposure of major bioactive ingredients in Chang-Kang-Fang, a multi-herb prescription for treatment of irritable bowel syndrome.

ETHNOPHARMACOLOGICAL RELEVANCE: Chang-Kang-Fang (CKF) is a traditional Chinese herbal formula used for treatment of irritable bowel syndrome (IBS) in China. Decoction is the administration form of CKF in clinical practice. Previously, CKF has been confirmed with activities of releasing pain and reversing disorders of intestinal propulsion. And alkaloids, monoglycosides, chromones were found as the main bioactive components potentially contributing to the efficacy of CKF. Polysaccharide was also a major constituent in CKF. But if and how polysaccharides influence the systemic exposure of bioactive components in CKF is unknown. AIM OF THE STUDY: In this study, we aimed to demonstrate the contribution of the co-existed polysaccharides on the systemic exposure of the major bioactive components from CKF in normal and IBS model rats. MATERIALS AND METHODS: An UPLC-TQ-MS with multiple reaction monitoring (MRM) scan method was developed and validated for quantifying six major small molecular bioactive ingredients of CKF in the plasma samples, including magnoflorine (MAG), berberine (BBR), albiflorin (ALB), paeoniflorin (PAE), 5-O-methylvisamminol (5-OM) and prim-O-glucosylcimifugin (POG). The rats received CKF decoction (CKF) and CKF small molecule portion (knockout of polysaccharides, CKFSM), respectively. IBS model rats were induced by daily bondage and gavage of Sennae Folium decoction (derived from the leaf of Cassia angustifolia Vahl). The effects of the co-existing polysaccharides on the pharmacokinetic parameters of six small molecular bioactive components in normal and IBS model rats were systematically evaluated. The potential gut microbiota involved mechanisms of the effects was validated by broad-spectrum antibiotic (ABX) treatment. RESULTS: The selectivity, precision, accuracy, recovery and matrix effect of the established quantification method were all within acceptable limits of biological sample. In normal rats, the co-existing polysaccharides significantly reduced the AUC(0-t) of MAG and PAE compared with CKFSM group. The Cmax and AUC(0-t) of other four compound were not influenced by co-existing polysaccharides. However, in IBS model rats, compared with CKFSM group, the Cmax and AUC(0-t) of the six ingredients significantly increased in CKF group. For CKF + ABX group, the Cmax of six ingredients decreased significantly when compared with CKF group, and the AUC(0-t) of MAG, BBR, ALB, PAE also reduced with significant differences. CONCLUSIONS: A reliable and sensitive UPLC-TQ-MS method was successfully developed and validated for evaluating influence of co-existing polysaccharides on pharmacokinetic behavior of six major small molecules components in CKF. The co-existing polysaccharides enhanced the systemic exposure of six bioactive small molecules in CKF under IBS pathological state potentially via gut microbiota involvement.