[Identification and visual analysis of ginsenosides in multi-steamed roots of Panax quinquefolium based on UPLC-Q-TOF-MS/MS and MALDI-MSI].

This study aims to investigate the component variations and spatial distribution of ginsenosides in Panax quinquefolium roots during repeated steaming and drying. Ultra performance liquid chromatography quadrupole time-of-flight tandem mass spectrometry(UPLC-Q-TOF-MS/MS) was employed to identify the ginsenosides in the root extract. Matrix-assisted laser desorption/ionization mass spectrometry imaging(MALDI-MSI) was employed to visualize the spatial distribution and spatiotemporal changes of prototype ginsenosides and metabolites in P. quinquefolium roots. The UPLC results showed that 90 ginsenosides were identified during the steaming process of the roots, and polar ginsenosides were converted into low polar or non-polar ginsenosides. The content of prototype ginsenosides decreased, while that of rare ginsenosides increased, which included 20(S/R)-ginsenoside Rg_3, 20(S/R)-ginsenoside Rh_2, and ginsenosides Rk_1, Rg_5, Rs_5, and Rs_4. MALDI-MSI results showed that ginsenosides were mainly distributed in the epidermis and phloem. As the steaming times increased, ginsenosides were transported to the xylem and medulla. This study provides fundamental information for revealing the changes of biological activity and pharmacological effect of P. quinquefolium roots that are caused by repeated steaming and drying and gives a reference for expanding the application scope of this herbal medicine.

Identification of raffinose family oligosaccharides in processed Rehmannia glutinosa Libosch using matrix-assisted laser desorption/ionization mass spectrometry image combined with machine learning.

RATIONALE: Currently, research on oligosaccharides primarily focuses on the physiological activity and function, with a few studies elaborating on the spatial distribution characterization and variation in the processing of Rehmannia glutinosa Libosch. Thus, imaging the spatial distributions and dynamic changes in oligosaccharides during the steaming process is significant for characterizing the metabolic networks of R. glutinosa. It will be beneficial to characterize the impact of steaming on the active ingredients and distribution patterns in different parts of the plant. METHODS: A highly sensitive matrix-assisted laser desorption/ionization mass spectrometry image (MALDI-MSI) method was used to visualize the spatial distribution of oligosaccharides in processed R. glutinosa. Furthermore, machine learning was used to distinguish the processed R. glutinosa samples obtained under different steaming conditions. RESULTS: Imaging results showed that the oligosaccharides in the fresh R. glutinosa were mainly distributed in the cortex and xylem. As steaming progressed, the tetra- and pentasaccharides were hydrolyzed and diffused gradually into the tissue section. MALDI-MS profiling combined with machine learning was used to identify the processed R. glutinosa samples accurately at different steaming intervals. Eight algorithms were used to build classification machine learning models, which were evaluated for accuracy, precision, recall, and F1 score. The linear discriminant analysis and random forest models performed the best, with prediction accuracies of 0.98 and 0.97, respectively, and thus can be considered for identifying the steaming durations of R. glutinosa. CONCLUSIONS: MALDI-MSI combined with machine learning can be used to visualize the distribution of oligosaccharides and identify the processed samples after steaming for different durations. This can enhance our understanding of the metabolic changes that occur during the steaming process of R. glutinosa; meanwhile, it is expected to provide a theoretical reference for the standardization and modernization of processing in the field of medicinal plants.

A Dietary Supplement Containing Micronutrients, Phosphatidylserine, and Docosahexaenoic Acid Counteracts Cognitive Impairment in D-Galactose-Induced Aged Rats.

At present, it is a trend to use dietary supplements to prevent age-related cognitive impairment. This study aimed to investigate the effects of a dietary supplement enriched with micronutrients, phosphatidylserine, and docosahexaenoic acid on cognitive performance using a D-galactose (D-gal) induced aging rat model. Seven-month-old male Sprague-Dawley rats were randomly divided into five groups, including the control group, D-gal model group, and low-dose (2 g/kg body weight), medium-dose (6 g/kg body weight), and high-dose (10 g/kg body weight) dietary supplement intervention groups, which were investigated for 13 weeks. The dietary supplement intervention was found to improve cognitive performance in Morris water maze test, increase superoxidase dismutase activity, reduce malondialdehyde activity, decrease tumor necrosis factor-α and interleukin-6 concentrations, inhibit the activation of astrocytes, and elevate brain-derived neurotrophic factor protein and mRNA expression in the brains of D-gal-induced aged rats. This dietary supplement customized for the aged can be applied to the restoration of cognitive performance by enhancing antioxidant and anti-neuroinflammatory abilities, up-regulating neurotrophic factors, and inhibiting the activation of astrocytes. These results will be useful for future studies focused on implementation in humans.

Dietary exposure and risk assessment of chlorinated paraffins in roots and rhizomes of traditional Chinese medicine herbs.

Traditional Chinese medicine (TCM) provides therapeutic and health care effects through dietary intake. Owing to the susceptibility of plants to contaminations, a risk assessment system is urgently needed to ensure the safe use of TCMs. In this study, the contamination levels and risks associated with the dietary intake of short-chain chlorinated paraffins (SCCPs) and medium-chain chlorinated paraffins (MCCPs) were investigated in six kinds of frequently-used TCM herbs. The concentrations varied from 144.4 to 1527.8 ng·g-1 dw for SCCPs and non-detect to 1214.1 ng·g-1 dw for MCCPs, with mean values of 551.5 and 259.8 ng·g-1 dw, respectively. A geographic distribution analysis indicated that the concentrations of CPs in TCMs were mainly associated with their levels of contamination in the ambient environment. Carbon atom-chlorine congener profiles of CPs were dominated by C10Cl7-8 and C14Cl7-8 congeners, accounting for 20.1% and 32.4% of the total SCCP and MCCP concentrations, respectively. Principal component analysis indicated that the TCM species might be the main factor influencing the accumulation of SCCPs congeners. Finally, a risk assessment reveals that the estimated daily intake and margin of exposure were far below levels that might pose a health risk, indicating an acceptable dietary intake of SCCPs and MCCPs in the studied TCMs. This is the first report of CPs in the TCM herbs and the obtained results are expected to aid in future evaluation of the quality of TCMs and ensuring diet and drug safety.

Analysis of dietary exposure and risk assessment of pesticide residues in roots and rhizomes of Chinese herbs.

Medicine food homologous (MFH) plants provide therapeutic and health care effects through diet. Thus, a risk assessment system for hazardous ingredient residues is urgently required to ensure their safe use. In this study, the pesticide contamination of six root and rhizome Chinese herbs, Ginseng Radix et Rhizoma, Panacis Quinquefolii Radix, Pseudostellariae Radix, Salviae Miltiorrhizae Radix et Rhizoma, Codonopsis Radix, and Glehniae Radix, and the risks associated with their intake were investigated. A total of 420 MFH plant samples collected from 22 provinces in China were tested, and 61 pesticides were detected in 413 samples. Multiple pesticide residues were detected in each MFH sample, with contents ranging from 0.0002 to 3.010 mg/kg dry weight. Carbendazim (≥47.14%) and propham (≥40%) were the most frequently detected pesticides. Risk assessment determined by hazard quotients indicated that the risks were acceptable, with no short- or long-term adverse health effects. However, considering the high incidence of residues and the detection of unregistered or even prohibited pesticides, strict supervision of soil quality and pesticide application (particularly cadusafos) in MFH plant cultivation are recommended to aid in monitoring MFH plant quality and ensuring diet and drug safety. PRACTICAL APPLICATION: Ensure the diet and drug safety of Chinese herbs.

Recent advances in research on vine tea, a potential and functional herbal tea with dihydromyricetin and myricetin as major bioactive compounds.

Vine tea has been used as an herbal tea by several ethnic minorities for hundreds of years in China. Flavonoids, a kind of indispensable component in a variety of nutraceutical, pharmaceutical and cosmetic applications, are identified to be the major metabolites and bioactive ingredients in vine tea. Interestingly, vine tea exhibits a wide range of significant bioactivities including anti-oxidant, anti-inflammatory, anti-tumor, antidiabetic, neuroprotective and other activities, but no toxicity. These bioactivities, to some extent, enrich the understanding about the role of vine tea in disease prevention and therapy. The health benefits of vine tea, particularly dihydromyricetin and myricetin, are widely investigated. However, there is currently no comprehensive review available on vine tea. Therefore, this report summarizes the most recent studies investigating bioactive constituents, pharmacological effects and possible mechanisms of vine tea, which will provide a better understanding about the health benefits and preclinical assessment of novel application of vine tea.

Discovery of 4-Arylindolines Containing a Thiazole Moiety as Potential Antitumor Agents Inhibiting the Programmed Cell Death-1/Programmed Cell Death-Ligand 1 Interaction.

Through specific structural modification of a 4-phenylindoline precursor, new 4-arylindolines containing a thiazole moiety were developed and found to be promising modulators of the programmed cell death-1 (PD-1)/programmed cell death-ligand 1 (PD-L1) axis. Compound A30 exhibited outstanding biochemical activity, with an IC50 of 11.2 nM in a homogeneous time-resolved fluorescence assay. In the cell-based assay, A30 significantly promoted IFN-γ secretion and rescued T-cell proliferation, which were inhibited by PD-1 activation. Furthermore, A30 showed favorable in vivo antitumor activity in a mouse 4T1 breast carcinoma model. Moreover, in mouse CT26 colon carcinoma models, A30 potently suppressed the growth of CT26/PD-L1 tumor but did not obviously affect the growth of CT26/vector tumor. The results of flow cytometry analysis indicated that A30 inhibited tumor growth by activating the immune microenvironment. We concluded that A30 is a new starting point for further development of PD-1/PD-L1 interaction inhibitors as antitumor agents.

Recent advances in research on vine tea,a potential and functional herbal tea with dihydromyricetin and myricetin as major bioactive compounds / 药物分析学报

Vine tea has been used as an herbal tea by several ethnic minorities for hundreds of years in China.Flavonoids,a kind of indispensable component in a variety of nutraceutical,pharmaceutical and cosmetic applications,are identified to be the major metabolites and bioactive ingredients in vine tea.Interest-ingly,vine tea exhibits a wide range of significant bioactivities including anti-oxidant,anti-inflammatory,anti-tumor,antidiabetic,neuroprotective and other activities,but no toxicity.These bioactivities,to some extent,enrich the understanding about the role of vine tea in disease prevention and therapy.The health benefits of vine tea,particularly dihydromyricetin and myricetin,are widely investigated.However,there is currently no comprehensive review available on vine tea.Therefore,this report summarizes the most recent studies investigating bioactive constituents,pharmacological effects and possible mechanisms of vine tea,which will provide a better understanding about the health benefits and preclinical assessment of novel application of vine tea.

Discovery andw biological evaluation of novel 6,7-disubstituted-4-(2-fluorophenoxy)quinoline derivatives possessing 1,2,3-triazole-4-carboxamide moiety as c-Met kinase inhibitors.

A series of 6,7-disubstituted-4-(2-fluorophenoxy)quinoline derivatives possessing 1,2,3-triazole-4-carboxamide moiety were designed, synthesized and evaluated for their in vitro biological activities against c-Met kinase and five typical cancer cell lines (A549, H460, HT-29, MKN-45 and U87MG). Most compounds showed moderate to excellent antiproliferative activity. In this study, a promising compound 34, with a c-Met IC50 value of 1.04nM, was identified as a multitargeted receptor tyrosine kinase inhibitor. The SAR analyses indicated that compounds with halogen group, especially fluoro group, at 4-position on the phenyl ring (moiety B) have potent antitumor activity, and methylation on the 5-atom linker played an important role in the c-Met enzymatic activity.

Discovery of novel 6,7-disubstituted-4-phenoxyquinoline derivatives bearing 5-(aminomethylene)pyrimidine-2,4,6-trione moiety as c-Met kinase inhibitors.

A series of novel quinoline derivatives bearing 5-(aminomethylene)pyrimidine-2,4,6-trione moiety were designed, synthesized, and evaluated for their c-Met kinase inhibitory activities and antiproliferative activities against 5 cancer cell lines (HT-29, H460, MKN-45, A549, and U87MG) in vitro. Most compounds showed moderate to excellent potency, with the most promising analogue 45 (c-Met half-maximal inhibitory concentration [IC50]=1.15 nM) showing high selectivity versus 5 other tyrosine kinases, VEGFR-2, Flt-3, PDGFR-ß, c-Kit, and EGFR. Structure-activity relationship studies indicated that electron-donating groups on the phenyl ring at the 3-position of pyrimidine-2,4,6-trione were required to increase the electron density on the 5-(aminomethylene)pyrimidine-2,4,6-trione moiety.

Curculigoside promotes osteogenic differentiation of bone marrow stromal cells from ovariectomized rats.

OBJECTIVES: Curculigoside, a natural compound isolated from the medicinal plant Curculigo orchioides has been reported to prevent bone loss in ovariectomized rats. However, the underlying molecular mechanisms are largely unknown. This study investigated the effects of curculigoside on proliferation and osteogenic differentiation of bone marrow stromal cells (BMSCs). METHODS: The toxicity, proliferation and osteogenic differentiation of BMSCs cultured with various concentrations (0 as control, 10, 100 and 500 µm) of curculigoside were measured by viability assay, MTT analysis, alkaline phosphatase (ALP) activity assay, alizarin red staining and mineralization assay, real-time PCR analysis on osteogenic genes including ALP, type I collagen (Col I), osteocalcin (OCN) and osteoprotegerin (OPG), runt-related transcription factor 2 (Runx2), as well as OPG enzyme-linked immunosorbent assay. KEY FINDINGS: No significant cytotoxicity was observed for BMSCs after supplementation with curculigoside. The proliferation of BMSCs was enhanced after administration of curculigoside, especially 100 µm curculigoside. Moreover, the osteogenic gene expression was significantly enhanced with 100 µm curculigoside treatment. Importantly, curculigoside significantly increased OPG secretion. CONCLUSIONS: The data indicate that curculigoside could promote BMSC proliferation and induce osteogenic differentiation of BMSCs. The most profound response was observed with 100 µm curculigoside. These findings may be valuable for understanding the mechanism of the effect of curculigoside on bone, especially in relation to osteoporosis.

Design, synthesis and antitumor activity of novel artemisinin derivatives using hybrid approach.

In an attempt to develop potent and selective anti-tumor agents, two novel series of artemisinin-chalcone hybrids were designed, synthesized and screened for their antitumor activities against HT-29, A549, MDA-MB-231, HeLa and H460 cell lines in vitro. Nearly all of the tested compounds showed significantly increased anti-tumor activity compared with the corresponding dihydroartemisinin (DHA). Most of the title compounds displayed good selectivity toward HT-29 and HeLa cell lines with IC50 values ranging from 0.09 to 0.85 µM. Among them, the most promising compound 9c (IC50) range of 0.09-0.93 µM) was 10.5- to 70-times more active than DHA (IC50 range of 5.6-15.6 µM) respectively.

Baseline and follow-up laboratory monitoring of cardiovascular medications.

BACKGROUND: Laboratory monitoring of medications is typically used to establish safety prior to drug initiation and to detect drug-related injury following initiation. It is unclear whether black box warnings (BBWs) as well as evidence- and consensus-based clinical guidelines increase the likelihood of appropriate monitoring. OBJECTIVE: To determine the proportion of patients newly initiated on selected cardiovascular medications with baseline assessment and follow-up laboratory monitoring and compare the prevalence of laboratory testing for drugs with and without BBWs and guidelines. METHODS: This cross-sectional study included patients aged 18 years or older from a large multispecialty group practice who were prescribed a cardiovascular medication (angiotensin converting enzyme inhibitors, angiotensin II receptor blockers, amiodarone, digoxin, lipid-lowering agents, diuretics, and potassium supplements) between January 1 and July 31, 2008. The primary outcome measure was laboratory test ordering for baseline assessment and follow-up monitoring of newly initiated cardiovascular medications. RESULTS: The number of new users of each study drug ranged from 49 to 1757 during the study period. Baseline laboratory test ordering across study drugs ranged from 37.4% to 94.8%, and follow-up laboratory test ordering ranged from 20.0% to 77.2%. Laboratory tests for drugs with baseline laboratory assessment recommendations in BBWs were more commonly ordered than for drugs without BBWs (86.4% vs 78.0%, p < 0.001). Drugs with follow-up monitoring recommendations in clinical guidelines had a lower prevalence of monitoring (33.1% vs 50.7%, p < 0.001). CONCLUSIONS: Baseline assessment of cardiovascular medication monitoring is variable. Quality measurement of adherence to BBW recommendations may improve monitoring.

Synthesis and in-vitro anti-hepatitis-B virus activity of 6H-[1]benzothiopyrano[4,3-b] quinolin-10-ols.

A series of 9-methoxy-6H-[1]benzothiopyrano[4,3-b]quinolin-10-ols with a Mannich side chain were synthesized and evaluated for their anti-Hepatitis B virus (HBV) activity in HepG2.2.15 cells. Some compounds showed significant anti-HBV activity with IC(50) values less than 41 microM. Among them, compound 9b was the most effective anti-HBV agent (IC(50) = 1.7 microM, SI = 60.3).

[Effects of different phosphorus substrates on the growth and phosphatase activity of Skeletonema costatum and Prorocentrum donghaiense].

The effects NaH2PO4, adenosine disodium triphosphate (ATP), glucose 6-phosphate (G-6-P) and sodium beta-glycerophosphate (G-P) on the growth and phosphatase activity of Skeletonema costatum and Prorocentrum donghaiense were studied. The results showed that both species could utilize both dissolved inorganic phosphate (DIP) and dissolved organic phosphorus (DOP), and DOP had more effects on the growth of two species than DIP. For S. costatum, after 8 days, the cell abundances of the four treatments (NaH2PO4, ATP, G-6-P and G-P) were 48 x 10(4), 73 x 10(4), 63 x 10(4) and 54 x 10(4) cells/mL, respectively; For P. donghaiense, after 10 days, the cell abundances of the four treatments were 8.7 x 10(4), 15.5 x 10(4), 12.4 x 10(4) and 9.5 x 10(4) cells/mL, respectively. On the first 3-4 days, the phosphatase activity of all treatments of the two species showed a decreasing trend, but different changes were observed for the different phosphorus substrate treatments in latter days. For the NaH2PO4 treatment, both the AP and AcP activity of two species increased from the fifth day onwards. For S. costatum, the AP activity of the ATP and G-6-P treatment groups showed no obvious changes and AcP activity had a slight increase from the fifth day to the eighth day, while the activity of G-P treatment had highest phosphatase activity which increased from the fifth day on. At the end of the experiment, the AP activity of the three DOP treatment groups (ATP, G-6-P and G-P) was 0.004 x 10(-5), 0.014 x 10(-5) and 0.029 x 10(-5) U/cell, respectively, and the AcP activity was 0.006 x 10(-5), 0.011 x 10(-5) and 0.018 x 10(-5) U/cell, respectively. For P. donghaiense, both the AP and AcP activity of the three DOP treatments had similar trends, i.e., ATP < G-6-P < G-P. Under the same nutrient conditions, S. costatum had a much higher phosphatase activity and could absorb P from the environment much faster than P. donghaiense.

[Comparative research on the effects of different nutrient concentrations on the photopigment content and photosynthesis of two bloom-forming species isolated from the Changjiang River estuary].

The contents of cellular chlorophyll a (Chl-a), chlorophyll c (Chl-c), total coloured carotenoids (TCC) and the photosynthesis of Skeletonema costatum and Prorocentrum donghaiense under different nutrient conditions were studied. The results showed that both species in the low nutrient concentration conditions had lower cellular Chl-a, Chl-c and TCC content than those in the high nutrient concentration conditions. When the initial N/P ratio was 16/1 while the concentrations were different, the two species had different photosynthetic rate responses. For S. costatum, the photosynthetic rate normalized by cell in the low nutrient concentration group (64 micromol/L N and 4 micromol/L P) was significantly lower than that in the high concentration group (256 micromol/L N and 16 micromol/L P) from the fifth day, and at the end of the experiment (on the seventh day), the photosynthetic rate was 0.031 x 10(-4) micromol x (cell x h)(-1) and 0.13 x 10(-4) micromol x (cell x h)(-1) respectively, while the photosynthetic rate normalized by Chl-a was 12.92 micromol x (microg x h)(-1) and 13.03 micromol x (microg x h)(-1) for the two groups respectively, and there was no significant difference between them; however, for P. donghaiense, the photosynthetic rates normalized by both cell and Chl-a in the low concentration conditions (64 micromol/L N and 4 micromol/L P) were significantly higher than those in the high concentration conditions (256 micromol/L N and 16 micromol/L P). For the two species, when P was sufficient, the low N concentration group (64 micromol/L N and 36 micromol/L P) and when N was sufficient, the low P concentration group (883 micromol/L N and 4 micromol/L P) had higher photosynthetic rates normalized by both cell and Chl-a than the high N concentration group (256 micromol/L N and 36 micromol/L P) and the high P concentration group (883 micromol/L N and 16 micromol/L P) respectively. There was a significant positive relationship between the photosynthetic rate and the intracellular P for both species, and P. donghaiense had higher cellular photopigment contents and photosynthetic rates than S. costatum under the same nutrient conditions. It could be inferred from their photosynthetic characteristics that P. donghaiense would survive better in low nutrient conditions compared with S. costatum.

[Fast determination of andrographolide and dehydroandrographolide in Xiaoyanlidan Tablets by microemulsion electrokinetic chromatography].

A microemulsion electrokinetic chromatography (MEEKC) method for the determination of andrographolide and dehydroandrographolide in Xiaoyanlidan Tablets was established. The MEEKC method involved the use of sodium dodecyl sulfate (SDS) as surfactant, ethyl acetate as organic solvent and 1-butanol as co-surfactant. The effects of several factors such as the acidity and concentration of borate buffer, SDS and 1-butanol contents were investigated. The optimized composition (by mass) of microemulsion system are 0.5% ethyl acetate--0.6% SDS--6.0% 1-butanol--92.9% 30 mmol/L sodium tetraborate buffer (pH 9.5). This system allowed a useful and reproducible separation of the analytes to be achieved in less than 6 min. The proposed method is a simple, fast and sensitive method for the determination of andrographolide and dehydroandrographolide in Xiaoyanlidan Tablets.

Rapid separation and determination of resibufogenin and cinobufagin in toad venom and Liushen tablet by beta-cyclodextrin modified micellar electrokinetic chromatography.

A rapid cyclodextrin modified micellar electrokinetic chromatography (CD-MEKC) method was proposed for the determination of resibufogenin and cinobufagin in the Chinese herbal extracts from toad venom and its medicinal preparation (Liushen tablet). The two components have the close structural similarity and similar hydrophobicity, which result in poor resolution in normal MEKC. The addition of neutral beta-CD to the MEKC system was found to improve the separation of the studied compounds. The effects of several CD-MEKC parameters on the resolutions were evaluated systematically. Based on the investigation, a background electrolyte solution consisting of 10 mM borate buffer adjusted to pH 8.5, 40 mM sodium dodecyl sulfate (SDS), 12 mM beta-CD and 10% (v/v) of methanol was found to be optimal conditions for the fast separation. The contents of resibufogenin and cinobufagin were successfully determined within 5 min, with satisfactory repeatability and recovery.