Saikosaponin B4 Suppression Cancer Progression by Inhibiting SW480 and SW620 Cells Proliferation via the PI3K/AKT/mTOR Pathway in Colon Cancer.

BACKGROUND: Colon cancer is a gastrointestinal malignancy with high incidence and poor prognosis. OBJECTIVE: Saikosaponin B4 (SSB4) is a monomeric component of the Traditional Chinese medicine (TCM), Bupleurum. The current study investigates the therapeutic effect and mechanisms of SSB4 in colon cancer. METHODS: The proliferation of two colon cancer cell lines, SW480 and SW620, were assessed using CCK8 and expression of regulatory molecules, including Bax, Caspase3, Caspase9, Cleaved Caspase3, Cleaved Caspase9 and Bcl2 by flow cytometry and Western blotting. RESULTS: Survival rates, assessed by CCK8, of SW480 and SW620 cells decreased significantly when the SSB4 concentration was in the range 12.5-50 µg/ml. Flow cytometry measurements indicated apoptosis rates of 55.07% ± 1.63% for SW480 cells and 33.07% ± 1.28% for SW620 cells treated with 25 µg/ml SSB4. Western blotting revealed upregulation of the proapoptotic proteins, Bax, Caspase3, Caspase9, Cleaved Caspase3 and Cleaved Caspase9, and downregulation of the anti-apoptotic protein, Bcl2, in the presence of SSB4. Network pharmacology and molecular docking predicted that the PI3K/Akt/mTOR pathway might be the main regulatory target for the antitumor effect of SSB4. Further Western blotting experiments showed that SSB4 downregulated (p < 0.01) expression of PI3K, Akt, mTOR and the phosphorylated proteins, P-PI3K, P-Akt and P-MTOR. Expression of PI3K, Akt and mTOR mRNA was found to be downregulated by SSB4 (P < 0.01) as the result of RT-PCR measurements. CONCLUSION: SSB4 is a potent anti-colon cancer agent. Its effects are likely to be mediated by suppression of the PI3K/AKT/mTOR pathway.

Hyperoside: A Review of Its Structure, Synthesis, Pharmacology, Pharmacokinetics and Toxicity.

Hyperoside is an active ingredient in plants, such as Hypericum monogynum in Hypericaceae, Crataegus pinnatifida in Rosaceae and Polygonum aviculare in Polygonaceae. Its pharmacologic effects include preventing cancer and protecting the brain, neurons, heart, kidneys, lung, blood vessels, bones, joints and liver, among others. Pharmacokinetic analysis of hyperoside has revealed that it mainly accumulates in the kidney. However, long-term application of high-dose hyperoside should be avoided in clinical practice because of its renal toxicity. This review summarises the structure, synthesis, pharmacology, pharmacokinetics and toxicity of hyperoside.

Thymus quinquecostatus Celak. ameliorates cerebral ischemia-reperfusion injury via dual antioxidant actions: Activating Keap1/Nrf2/HO-1 signaling pathway and directly scavenging ROS.

BACKGROUND: Thymus quinquecostatus Celak. has been widely used as a spice and a folk medicine for relieving exterior syndrome and alleviating pain in China. PURPOSE: To explore the protective effects and the underlying mechanism against cerebral ischemia-reperfusion injury (CIRI) of the T. quinquecostatus combining with its chemical composition. STUDY DESIGN AND METHODS: High-polar extract (HPE) was extracted from T. quinquecostatus and polyphenols in HPE were enriched to obtain polyphenol-rich fraction (PRF) using Macroporous resin. The free radicals and zebrafish embryos were used to compare the antioxidant activities of HPE and PRF in vitro and in vivo. Then, the transient middle cerebral artery occlusion (tMCAO) model was established in rats. Neurological deficit score, infarction rate, morphology and apoptosis of neurons were examined to investigate the protective effects of PRF on CIRI. The mRNA and protein levels of nuclear factor erythroid 2-related factor 2 (Nrf2) and hemeoxygenase-1 (HO-1) and the activities of downstream antioxidant enzymes in ischemia tissues were determined to clarify the underlying mechanisms. Also, reactive oxygen species (ROS) level in zebrafish embryos were detected after incubation with PRF for a short time (2 h) to investigate whether PRF could directly eliminate free radicals. Finally, chemical composition of PRF were analyzed to investigate the material basis for antioxidant activity and anti-CIRI effect. RESULTS: Compared with HPE, PRF showed stronger antioxidant activities. PRF exhibited obvious protective effects including ameliorating neurological deficit, lowering infarction rate, and improving the cellular morphology in hippocampus CA1 and cortex after tMCAO. TUNEL staining suggested PRF dose-dependently improved the apoptosis of the neurons in ischemic cortex. RT-qPCR and Western Blot results suggested that PRF regulated oxidative stress (OS) via activating the Keap1/Nrf2/HO-1 signaling pathway. Also, PRF could directly scavenge excessive ROS in zebrafish embryos after a short-time PRF incubation. The anti-CIRI effect might be primarily attributed to the abundant polyphenols in PRF, including flavonoids, polymethoxylated flavonoids, flavonoid glycosides, and phenolic acids. CONCLUSION: T. quinquecostatus contains abundant polyphenols and exhibited a good protective effect against CIRI via dual antioxidant mechanisms, providing a reference for further research and application for this plant.

Phlomoides umbrosa (Turcz.) Kamelin & Makhm: A review of its traditional uses, botany, phytochemistry, pharmacology and clinical research.

ETHNOPHARMACOLOGICAL RELEVANCE: Phlomoides umbrosa (Turcz.) Kamelin & Makhm (P. umbrosa, Lamiaceae) is also known as "Caosu" in China and "Han Sok-Dan" in Korea. It has been used as a traditional medicine for hundreds of years. This plant is not only as a traditional medicine to alleviate diseases such as colds, arthritis, osteoporosis, but also as a food additive. AIM OF THE STUDY: This review provides up-to-date investigations of this plant, including its botany, traditional uses, pharmacology, phytochemistry, clinical research, cytotoxicity, and safety evaluation. The possible purposes and perspectives for future research of P. umbrosa are also discussed. MATERIALS AND METHODS: Information on the studies of P. umbrosa is collected from scientific journals and reports via library and electronic data search (PubMed, Baidu Academic, Google Scholar, Science Direct, ACS, Web of Science, and CNKI). Meanwhile, it is also obtained from published works of folk records, ethnopharmacological literature, Ph.D. and Masters Dissertation. RESULTS: Phytochemical research reveals that this plant contains triterpenoids, iridoids, phenylethanoids, flavonoids, essential oil, microelement, etc. The extract of P. umbrosa exhibits extensive pharmacological activities including anti-osteoporosis, anti-allergic, anti-bacterial, anti-nociceptive, anti-inflammatory, anti-oxidant, anti-cancer. Almost no obvious toxicity or side effects is observed and recorded for P. umbrosa. CONCLUSIONS: This review summarizes traditional uses, botany, pharmacology, phytochemistry, clinical research, cytotoxicity, and safety evaluation of P. umbrosa, and presents the constituents and their corresponding chemical structures found in P. umbrosa comprehensively for the first time. Meanwhile, modern pharmacological studies also are extensively investigated at present. It is worth mentioning that P. umbrosa promotes children's growth as well as the application of clinical research. Although there are clinical studies on P. umbrosa, its pharmacokinetics needs to be further elucidated. Besides, P. umbrosa is also limited in identifying active compounds and clarifying pharmacological mechanisms. Similarly, modern researches on the traditional application of P. umbrosa should also be urgently confirmed, such as treatment of fractures and hemostasis. It is believed that this review will provide a theoretical basis and valuable data for future in-depth research and application.

Traditional Uses, Chemical Constituents and Biological Activities of Plants from the Genus Thymus.

The genus Thymus (Lamiaceae) comprises about 214 species throughout the world, mainly found in North Africa, Europe, and temperate Asia zone. They are traditionally used as food additives and folk medicines. This review comprehensively summarizes information about traditional uses, chemical constituents, and biological activities of this genus and provides recommendations for future investigations. All information was gathered from scientific databases including Google Scholar, Sci-Finder, Web of Science, ScienceDirect, and CNKI. Volatile oils are the most concerned constituents of this genus. Flavonoids, phenylpropanoids, tannins, organic acids, terpenoids, and phytosterols were also summarized. This genus plants possessed a variety of activities including antimicrobial, antioxidant, anti-inflammatory, cytotoxic, analgesic, and antidiabetic. In brief, this review will be helpful to provide valuable data for explorations and create more interests towards Thymus genus in the future.

Recovery of extra-heavy oil and minerals from carbonate asphalt rocks by reactive extraction.

Quite different from the Canadian oil sands, the Indonesian asphalt rocks proved to be carbonate unconventional oil ores. The strong interactions between asphalt and minerals make water-based extraction work poorly in separating this kind of ore. Herein, a reactive extraction process has been proposed to separate asphalt and mineral solids from the ores through dissolving the mineral solids (i.e., carbonate minerals, metal oxides, etc.) by acids (formic acid). It is evidenced that most of the asphalt could be recovered and collected on the top of the solution by generated CO2. What's more, the unreacted formic acid could be recycled in this process. The dissolved metal ions could be efficiently recovered to obtain different by-products by chemical settling and crystallization. The amount of residual solids settled at the bottom of the reactor is very small. Further tests show that the reaction efficiency is highly dependent on the operational conditions, including temperature, stirring rate, acid dosage, concentration of acid, etc. It is also found that the reaction could allow minerals to be redistributed in different phases. Although some metal elements could be dissolved into solution, elements such as Fe, Al, S, Si, and Ti are observed to accumulate in asphalt froth. In addition to reacting with minerals, formic acid is also found to reduce asphalt viscosity. This reduction improves the reaction efficiency. Based on primary evaluations, the above findings suggest that the reactive extraction would be a potential process to exploit the Indonesian asphalt rocks (or other similar ores) due to its full recovery to all materials.

Discrimination of three Ephedra species and their geographical origins based on multi-element fingerprinting by inductively coupled plasma mass spectrometry.

Discrimination of species and geographical origins of traditional Chinese medicine (TCM) is essential to prevent adulteration and inferior problems. We studied Ephedra sinica Stapf, Ephedra intermedia Schrenk et C.A.Mey. and Ephedra przewalskii Bge. to investigate the relationship between inorganic element content and these three species and their geographical origins. 38 elemental fingerprints from six major Ephedra-producing regions, namely, Inner Mongolia, Ningxia, Gansu, Shanxi, Shaanxi, and Sinkiang, were determined to evaluate the importance of inorganic elements to three species and their geographical origins. The contents of 15 elements, namely, N, P, K, S, Ca, Mg, Fe, Mn, Na, Cl, Sr, Cu, Zn, B, and Mo, of Ephedra samples were measured using inductively coupled plasma mass spectroscopy. Elemental contents were used as chemical indicators to classify species and origins of Ephedra samples using a radar plot and multivariate data analysis, including hierarchical cluster analysis (HCA), principal component analysis (PCA), and discriminant analysis (DA). Ephedra samples from different species and geographical origins could be differentiated. This study showed that inorganic elemental fingerprint combined with multivariate statistical analysis is a promising tool for distinguishing three Ephedra species and their geographical origins, and this strategy might be an effective method for authenticity discrimination of TCM.

Multi-Response Extraction Optimization Based on Anti-Oxidative Activity and Quality Evaluation by Main Indicator Ingredients Coupled with Chemometric Analysis on Thymus quinquecostatus Celak.

Thymus quinquecostatus Celak is a species of thyme in China and it used as condiment and herbal medicine for a long time. To set up the quality evaluation of T. quinquecostatus, the response surface methodology (RSM) based on its 2,2-Diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity was introduced to optimize the extraction condition, and the main indicator components were found through an UPLC-LTQ-Orbitrap MSn method. The ethanol concentration, solid-liquid ratio, and extraction time on optimum conditions were 42.32%, 1:17.51, and 1.8 h, respectively. 35 components having 12 phenolic acids and 23 flavonoids were unambiguously or tentatively identified both positive and negative modes to employ for the comprehensive analysis in the optimum anti-oxidative part. A simple, reliable, and sensitive HPLC method was performed for the multi-component quantitative analysis of T. quinquecostatus using six characteristic and principal phenolic acids and flavonoids as reference compounds. Furthermore, the chemometrics methods (principal components analysis (PCA) and hierarchical clustering analysis (HCA)) appraised the growing areas and harvest time of this herb closely relative to the quality-controlled. This study provided full-scale qualitative and quantitative information for the quality evaluation of T. quinquecostatus, which would be a valuable reference for further study and development of this herb and related laid the foundation of further study on its pharmacological efficacy.

Synthesis and application of hydrophilically-modified Fe3O4 nanoparticles in oil sands separation.

Nanoparticles have been reported to be a promising candidate for the separation of heavy oil from its host rock's surface. These nanoparticles (NPs) are often dispersed and stabilized in the solution by some surfactants during the unconventional oil ores processing. Herein, the PEG600-KH560 (PK) has been grafted onto Fe3O4 NP surfaces, obtaining a kind of hydrophilically-modified recyclable nanoparticle. Results show that these NPs (averaged at around 16 nm for single sphere) could be well dispersed in water (no settling in 72 h), forming PK-Fe3O4 nanofluids (NFs) at 0.2 wt%. These PK-Fe3O4 NFs are found to be able to be quickly separated from the dispersions by an external magnetic field, and returning back to stable NFs when the magnetic field disappears and by shaking. The PK-Fe3O4 NFs have been further used for the enhancement of heavy oil recovery from oil sands. The floatation results show that the PK-Fe3O4 NFs could improve oil recovery by at least 12% compared with the traditional hot water extraction process (HWEP). After the extraction, up to 70% of the PK-Fe3O4 NPs could be directly recycled from the solution for further use. The rest of the NPs are left in the oil phase and attached on the residual solid surface. However, the efficiency of the PK-Fe3O4 NPs is found to be decreased when the recycling times exceed 5 due to the adsorption of oil components. A mechanistic study shows that the hydrophilic PK-Fe3O4 NPs could be adsorbed on the mineral surface, making the surface more hydrophilic. The hydrophilic surface and the agitation disturbance helps the liberation process of bitumen from the solid surfaces. On the other hand, when adding the PK-Fe3O4 NPs into the heavy oil-water system, the oil-water interface is found to be highly modified by the NPs, resulting in significant reduction of the oil-water interfacial tension. The above findings suggest that the PK-Fe3O4 NPs combined the surface-active role (surfactant) and the nano-size role (adsorption) together, which facilitates its role in oil sands separation.

Discrimination of geographical origin of cultivated Polygala tenuifolia based on multi-element fingerprinting by inductively coupled plasma mass spectrometry.

Inorganic elements are important components of medicinal herbs, and provide valuable experimental evidence for the quality evaluation and control of traditional Chinese medicine (TCM). In this study, to investigate the relationship between the inorganic elemental fingerprint and geographical origin identification of cultivated Polygala tenuifolia, 41 elemental fingerprints of P. tenuifolia from four major polygala-producing regions (Shanxi, Hebei, Henan, and Shaanxi) were evaluated to determine the importance of inorganic elements to cultivated P. tenuifolia. A total of 15 elemental (B, Ca, Cl, Cu, Fe, K, Mg, Mn, Na, N, Mo, S, Sr, P, and Zn) concentrations of cultivated P. tenuifolia were measured using inductively coupled plasma mass spectroscopy (ICP-MS). The element composition samples were classified by radar plot, elemental fingerprint, and multivariate data analyses, such as hierarchical cluster analysis (HCA), principle component analysis (PCA), and discriminant analysis (DA). This study shows that radar plots and multivariate data analysis can satisfactorily distinguish the geographical origin of cultivated P. tenuifolia. Furthermore, PCA results revealed that N, Cu, K, Mo, Sr, Ca, and Zn are the characteristic elements of cultivated P. tenuifolia. Therefore, multi-element fingerprinting coupled with multivariate statistical techniques can be considered an effective tool to discriminate geographical origin of cultivated P. tenuifolia.

Oral fast-dissolving films containing lutein nanocrystals for improved bioavailability: formulation development, in vitro and in vivo evaluation.

Lutein is widely used as diet supplement for prevention of age-related macular degeneration. However, the application and efficacy of lutein in food and nutritional products has been hampered due to its poor solubility and low oral bioavailability. This study aimed to develop and evaluate the formulation of oral fast-dissolving film (OFDF) containing lutein nanocrystals for enhanced bioavailability and compliance. Lutein nanocrystals were prepared by anti-solvent precipitation method and then encapsulated into the films by solvent casting method. The formulation of OFDF was optimized by Box-Behnken Design (BBD) as follows: HPMC 2.05% (w/v), PEG 400 1.03% (w/v), Cremophor EL 0.43% (w/v). The obtained films exhibited uniform thickness of 35.64 ± 1.64 µm and drug content of 0.230 ± 0.003 mg/cm2 and disintegrated rapidly in 29 ± 8 s. The nanocrystal-loaded films with reconstituted particle size of 377.9 nm showed better folding endurance and faster release rate in vitro than the conventional OFDFs with raw lutein. The microscope images, thermograms, and diffractograms indicated that lutein nanocrystals were highly dispersed into the films. After administrated to SD rats, t max was decreased from 3 h for oral solution formulation to less than 0.8 h for OFDF formulations, and C max increased from 150 ng/mL for solution to 350 ng/mL for conventional OFDF or 830 ng/mL for nanocrystal OFDF. The AUC 0-24h of conventional or nanocrystal OFDF was 1.37 or 2.08-fold higher than that of the oral solution, respectively. These results suggested that drug nanocrystal-loaded OFDF can be applied as a promising approach for enhanced bioavailability of poor soluble drugs like lutein.

Influence of soil and hydrocarbon properties on the solvent extraction of high-concentration weathered petroleum from contaminated soils.

Petroleum ether was used to extract petroleum hydrocarbons from soils collected from six oil fields with different history of exploratory and contamination. It was capable of fast removing 76-94 % of the total petroleum hydrocarbons including 25 alkanes (C11-C35) and 16 US EPA priority polycyclic aromatic hydrocarbons from soils at room temperature. The partial least squares analysis indicated that the solvent extraction efficiencies were positively correlated with soil organic matter, cation exchange capacity, moisture, pH, and sand content of soils, while negative effects were observed in the properties reflecting the molecular size (e.g., molecular weight and number of carbon atoms) and hydrophobicity (e.g., water solubility, octanol-water partition coefficient, soil organic carbon partition coefficient) of hydrocarbons. The high concentration of weathered crude oil at the order of 10(5) mg kg(-1) in this study was demonstrated adverse for solvent extraction by providing an obvious nonaqueous phase liquid phase for hydrocarbon sinking and increasing the sequestration of soluble hydrocarbons in the insoluble oil fractions during weathering. A full picture of the mass distribution and transport mechanism of petroleum contaminants in soils will ultimately require a variety of studies to gain insights into the dynamic interactions between environmental indicator hydrocarbons and their host oil matrix.

Solvent extraction for heavy crude oil removal from contaminated soils.

A new strategy of heavy crude oil removal from contaminated soils was studied. The hexane-acetone solvent mixture was used to investigate the ability of solvent extraction technique for cleaning up soils under various extraction conditions. The mixtures of hexane and acetone (25 vol%) were demonstrated to be the most effective in removing petroleum hydrocarbons from contaminated soils and approx 90% of saturates, naphthene aromatics, polar aromatics, and 60% of nC(7)-asphaltenes were removed. Kinetic experiments demonstrated that the equilibrium was reached in 5 min and the majority of the oil pollutants were removed within 0.5 min. The effect of the ratio between solvent and soil on the extraction efficiency was also studied and results showed that the efficiency would increase following the higher solvent soil ratio. Then the multistage continuous extraction was considered to enhance the removal efficiency of oil pollutants. Three stages crosscurrent and countercurrent solvent extraction with the solvent soil ratio 6:1 removed 97% oil contaminants from soil. Clearly the results showed that the mixed-solvent of hexane and acetone (25 vol%) with character of low-toxic, acceptable cost and high efficiency was promising in solvent extraction to remove heavy oil fractions as well as petroleum hydrocarbons from contaminated soils.

Chemical composition and antioxidant activities of extracts from Apocyni Veneti Folium.

An expeditious and effective HPLC-UV method has been developed for the simultaneous determination of seven major flavonoids in Apocyni Veneti Folium (AVF) extract. The chemical profile of seven flavonoids, including quercetin-3-O-ß-D-glc(2 → 1)-ß-D-glucoside, rutin, isoquercetin, kaempferol-3-O-ß-D-glucoside, quercetin-3-O-(6″-O-malonyl)-ß-D-glucoside, quercetin and kaempferol was acquired by HPLC-UV. The analysis was performed on a Diamosil C18 analytical column with a gradient solvent system of acetonitrile-0.1% aqueous acetic acid. Full validation of the method was carried out (linearity, reproducibility, repeatability, accuracy and limit of detection). The results indicated that the contents of investigated flavonoids in Apocyni Veneti Folium varied significantly from habitat to habitat, with contents ranging from 0.01 to 5.57 mg g⁻¹. The antioxidant activity results demonstrate that the seven flavonoids showed great efficiency in scavenging DPPH radicals. The high content of flavonoid components of AVF could be responsible for its high antioxidant activity. This study provides powerful evidence for the relationship between the chemical ingredients of and bioactivity in AVF.

[Biological toxicity effect of petroleum contaminated soil before and after physicochemical remediation].

Toxicity analysis was studied from using seed germination as an ecological indicator, and the earthworm was considered as a suitable biomonitor animal to determine the ecological hazard of polluted soil. The main results are as follows: These crop seeds have significantly different responses to petroleum pollution. Compared with those plants in clean soil, the germination of most crop seeds planted in contaminated soils is obviously inhabited. Soybean, horse bean and maize are the crop affected most adversely. Fortunately, strong endurance is observed for green soybean under 4 different levels of petroleum pollution, and the seed germination rate are all above 90%. When exposed to pollutants, earthworms could be changed obviously on the level of physiology. That might affect the survival and growth capacity of earthworms, and changed population finally. In high petroleum contaminated soil (concentration of petroleum > 30 000 mg/kg) earthworms can only survive about 5 days. The results suggest that petroleum pollution has great poison to earthworms and can kill earthworms finally. Because pollutants make them dehydrate. Even on the low pollution level, the survival time of earthworm is still very short (3 d or so) in the treated petroleum-contaminated soil. Because after a petroleum ether-treated, the nutrients of soil are disposed with the oil, and the organic matter and other nutrients of the soil have a great impact on the survival of earthworms.

In vitro and in vivo evaluation of actively targetable nanoparticles for paclitaxel delivery.

The aim of the present work was to assess the merits of an actively targetable nanoparticles (ATN), PEG-coated biodegradable polycyanoacrylate nanoparticles (PEG-nanoparticles) conjugated to transferrin, for paclitaxel delivery. PEG-nanoparticles loading paclitaxel were prepared by solvent evaporation technique in advance. ATN were prepared by coupling of transferrin to PEG-nanoparticles. The results showed that the average encapsulation efficiency of ATN was 93.4+/-3.6% with particle size (101.4+/-7.2 nm) and zeta-potential (-13.6+/-1.1 mV). The paclitaxel loaded ATN exhibited a low burst effect with about only 16.2% drug release within the first phase. Subsequently, paclitaxel release profiles displayed a sustained release phase. The amount of cumulated paclitaxel release over 30 days was 81.6%. ATN exhibited a markedly delayed blood clearance in mice, and the paclitaxel level from ATN remained much higher at 24 h compared with that of free drug from paclitaxel injection. The distribution profiles of ATN in S-180 solid tumor-bearing mice after intravenous administration showed the tumor accumulation of paclitaxel increase with time, and the paclitaxel concentration in tumor was about 4.8 and 2.1 times higher than those from paclitaxel injection and PEG-nanoparticles at 6 h after intravenous injection. For mice treated with 20 mg/kg x 5 of ATN, the decrease in body weight was limited within 4% of the initial weight at 5 days after the final administration, and tumor regression was significantly observed with complete tumor regression for five out of nine mice. The tumor burden with ATN-treated mice was much smaller compared with free paclitaxel or NTN-treated mice. In addition, the life span of tumor-bearing mice was significantly increased when they were treated with ATN, in particular, three mice survived over 60 days. Thus, PEG-coated biodegradable polycyanoacrylate nanoparticles conjugated to transferrin could be an effective carrier for paclitaxel delivery.