Rosa roxburghii Fruit Extracts Upregulate Telomerase Activity and Ameliorate Cell Replicative Senescence.

Anti-aging functional foods benefit the elderly. Telomeres are chromosomal ends that maintain genome stability extended by telomerase catalytic subunit TERT. Due to the end-replication problem, telomeres shorten after each cell cycle without telomerase in most human cells, and eventually the cell enters the senescence stage. Natural products can attenuate the aging process by increasing telomerase activity, such as TA-65. However, TA-65 is expensive. Other Chinese natural products may achieve comparable effects. Here, we found that Rosa roxburghii fruit extracts effectively increase TERT expression and telomerase activity in cultured human mesenchymal stem cells. Both R. roxburghii fruit extracts obtained by freeze-drying and spray-drying increased the activity of telomerase. R. roxburghii fruit extracts were able to reduce reactive oxygen species levels, enhance superoxide dismutase activity, and reduce DNA damage caused by oxidative stress or radiation. R. roxburghii fruit extracts promoted cell proliferation, improved senescent cell morphology, delayed replicative cellular senescence, attenuated cell cycle suppressors, and alleviated the senescence-associated secretory phenotype. Transcriptome and metabolic profiling revealed that R. roxburghii fruit extracts promote DNA replication and telomere maintenance pathways and decrease triglyceride levels. Overall, we provide a theoretical basis for the application of R. roxburghii fruit as an anti-aging product.

Application for Identifying the Origin and Predicting the Physiologically Active Ingredient Contents of Gastrodia elata Blume Using Visible-Near-Infrared Spectroscopy Combined with Machine Learning.

Gastrodia elata (G. elata) Blume is widely used as a health product with significant economic, medicinal, and ecological values. Due to variations in the geographical origin, soil pH, and content of organic matter, the levels of physiologically active ingredient contents in G. elata from different origins may vary. Therefore, rapid methods for predicting the geographical origin and the contents of these ingredients are important for the market. This paper proposes a visible-near-infrared (Vis-NIR) spectroscopy technology combined with machine learning. A variety of machine learning models were benchmarked against a one-dimensional convolutional neural network (1D-CNN) in terms of accuracy. In the origin identification models, the 1D-CNN demonstrated excellent performance, with the F1 score being 1.0000, correctly identifying the 11 origins. In the quantitative models, the 1D-CNN outperformed the other three algorithms. For the prediction set of eight physiologically active ingredients, namely, GA, HA, PE, PB, PC, PA, GA + HA, and total, the RMSEP values were 0.2881, 0.0871, 0.3387, 0.2485, 0.0761, 0.7027, 0.3664, and 1.2965, respectively. The Rp2 values were 0.9278, 0.9321, 0.9433, 0.9094, 0.9454, 0.9282, 0.9173, and 0.9323, respectively. This study demonstrated that the 1D-CNN showed highly accurate non-linear descriptive capability. The proposed combinations of Vis-NIR spectroscopy with 1D-CNN models have significant potential in the quality evaluation of G. elata.

Isolation of Chalcomoracin as a Potential α-Glycosidase Inhibitor from Mulberry Leaves and Its Binding Mechanism.

Diabetes is a chronic metabolic disease, whereas α-glucosidases are key enzymes involved in the metabolism of starch and glycogen. There is a long history of the use of mulberry leaf (the leaf of Morus alba) as an antidiabetic herb in China, and we found that chalcomoracin, one of the specific Diels-Alder adducts in mulberry leaf, had prominent α-glucosidase inhibitory activity and has the potential to be a substitute for current hypoglycemic drugs such as acarbose, which have severe gastrointestinal side effects. In this study, chalcomoracin was effectively isolated from mulberry leaves, and its α-glucosidase inhibition was studied via enzymatic kinetics, isothermal titration (ITC) and molecular docking. The results showed that chalcomoracin inhibited α-glucosidase through both competitive and non-competitive manners, and its inhibitory activity was stronger than that of 1-doxymycin (1-DNJ) but slightly weaker than that of acarbose. ITC analysis revealed that the combination of chalcomoracin and α-glucosidase was an entropy-driven spontaneous reaction, and the molecular docking results also verified this conclusion. During the binding process, chalcomoracin went into the "pocket" of α-glucosidase via hydrophobic interactions, and it is linked with residues Val544, Asp95, Ala93, Gly119, Arg275 and Pro287 by hydrogen bonds. This study provided a potential compound for the prevention and treatment of diabetes and a theoretical basis for the discovery of novel candidates for α-glycosidase inhibitors.

Assessment of Amentoflavone Loaded Sub-Micron Particle Preparation using Supercritical Antisolvent for its Antitumor Activity.

INTRODUCTION: The amentoflavone (AMF) loaded polymeric sub-micron particles were prepared using supercritical antisolvent (SAS) technology with the aim of improving the anticancer activity of AMF. METHODS: Zein and phospholipid mixtures composed of Hydrogenated Phosphatidylcholine (HPC) and egg lecithin (EPC) were used as carrier materials and, the effects of carrier composition on the product morphology and drug release behavior were investigated. When the mass ratio of Zein/HPC/ EPC was 7/2/1, the AMF loaded particles were spherical shape and sub-micron sized around 400 nm, with a drug load of 4.3±0.3 w% and entrapment efficacy of 87.8±1.8%. The in vitro drug release assay showed that adding EPC in the wall materials could improve the dispersion stability of the released AMF in an aqueous medium, and the introduction of HPC could accelerate the drug release speed. RESULTS: MTT assay demonstrated that AMF-loaded micron particles have an improved inhibitory effect on A375 cells, whose IC50 was 37.39µg/ml, compared with that of free AMF(130.2µg/ml). CONCLUSION: It proved that the AMF loaded sub-micron particles prepared by SAS were a prospective strategy to improve the antitumor activity of AMF, and possibly promote the clinical use of AMF preparations.

Long-Term Antibacterial Film Nanocomposite Incorporated with Patchouli Essential Oil Prepared by Supercritical CO2 Cyclic Impregnation for Wound Dressing.

Biocompatible skin wound dressing materials with long-term therapeutic windows and anti-infection properties have attracted great attention all over the world. The cooperation between essential oil and non-toxic or bio-based polymers was a promising strategy. However, the inherent volatility and chemical instability of most ingredients in essential oils make the sustained pharmacological activity of essential oil-based biomaterials a challenge. In this study, a kind of film nanocomposite loaded with patchouli essential oil (PEO-FNC) was fabricated. PEO-loaded mesoporous silica nanoparticles (PEO-MSNs) with drug load higher than 40 wt% were firstly prepared using supercritical CO2 cyclic impregnation (SCCI), and then combined with the film matrix consisting of polyvinyl alcohol and chitosan. The morphology of PEO-MSNs and PEO-FNC was observed by transmission and scanning electron microscope. The mechanical properties, including hygroscopicity, tensile strength and elongation at break (%), were tested. The release behavior of PEO from the film nanocomposite showed that PEO could keep releasing for more than five days. PEO-FNC exhibited good long-term (>48 h) antibacterial effect on Staphylococcus aureus and non-toxicity on mouse fibroblast (L929 cells), making it a promising wound dressing material.

Platinum (II)-doped graphitic carbon nitride with enhanced peroxidase-like activity for detection of glucose and H2O2.

A platinum (II)-doped graphitic carbon nitride (Pt2+@g-C3N4) nanozyme was prepared in this work. Compared to g-C3N4, Pt2+@g-C3N4 shows enhanced peroxidase-like activity. According to DFT calculations and ESR measurements, the Pt on the surface of g-C3N4 accelerates the decomposition of H2O2 into OH and consequently accelerates TMB oxidation. Furthermore, as a proof of concept, the obtained Pt2+2.30@g-C3N4 was applied in the detection of glucose with an LOD of 0.01 mM. The reproducibility and precision tests of the glucose detection method based on Pt2+2.30@g-C3N4 show that this method is reliable and feasible for the detection of glucose.

An efficient, green, and easy-to-scale-up strategy for target-oriented isolating cadinene sesquiterpenoids from Eupatorium adenophorum Spreng.

A green and efficient strategy was established and optimized for target-oriented extraction, enrichment and separation of cadinene sesquiterpenoids from Eupatorium adenophorum Spreng., using the combination of supercritical fluid extraction, molecular distillation, and industrial preparative chromatography for the first time. The extraction conditions of supercritical fluid extraction were initially optimized by orthogonal experimental design. Under the optimum conditions, the contents of 9-oxo-10,11-dehydroageraphorone and 10Hß-9-oxo-ageraphorone, which were 55.00% and 6.01%, respectively, were much higher than conventional extraction methods. Then, the molecular distillation enrichment method was established and investigated by response surface methodology technology, which showed strong specificity for enriching target compounds and removing impurities from crude extracts. Under the optimum conditions of molecular distillation, total contents of cadinene sesquiterpenoids were increased to 89.19%. Finally, a total of 146 mg of 9-oxo-10,11-dehydroageraphorone and 29 mg of 10Hß-9-oxo-ageraphorone were easily obtained by industrial preparative chromatography, from 200 mg of distillation fraction, with purities over 99%. The contents of target components were analyzed by HPLC, and structures of them were identified by high-resolution MS, 1 H-NMR, and 13 C-NMR spectroscopy. These results indicate that it is a simple, effective, and eco-friendly strategy, which is easily converted into industrial scale.

Zein/MCM-41 Nanocomposite Film Incorporated with Cinnamon Essential Oil Loaded by Modified Supercritical CO2 Impregnation for Long-Term Antibacterial Packaging.

Antimicrobial medicine and food packages based on bio-based film containing essential oils have attracted great attention worldwide. However, the controlled release of essential oils from these film nanocomposites is still a big challenge. In this study, a long-term antibacterial film nanocomposite composed of zein film and cinnamon essential oil (CEO) loaded MCM-41 silica nanoparticles was prepared. The CEO was loaded into MCM-41 particles via modified supercritical impregnation efficiently with a high drug load (>40 wt%). The morphologies of the prepared nanoparticles and film nanocomposite were characterized by a scanning electron microscope. The release behaviors of CEO under different temperatures, high humidity, continuous illumination and in phosphate buffer solution (PBS) solution were investigated. The results showed that the film nanocomposite had an outstanding release-control effect. The addition of MCM-41 nanoparticles also improved the mechanical properties of zein films. The antibacterial effect of CEO was significantly prolonged by the film nanocomposite; indicating the CEO film nanocomposite fabricated via modified supercritical CO2 impregnation was a potential long-term antibacterial medicine or food package material.

Effects of combined extracts of Lepidium meyenii and Allium tuberosum Rottl. on erectile dysfunction.

BACKGROUND: Sexual problems are widespread and adversely affect the interpersonal relationships and the quality of life. Currently, synthetic drugs improving sexual function are available, but expenditures for such agents are extremely high. To discover relatively inexpensive, widely available and effective natural drugs, we identified a combined extracts from Lepidium meyenii (maca) root and Allium tuberosum Rottl. (Chinese chive) seed, assessed the effects of this combined extracts on erectile dysfunction, and explored its potential mechanisms. METHODS: The extracts were obtained via supercritical fluid extraction. Male BALB/c mice received doses of extract from single plant or the combined extracts (200 mg/kg) by gastric gavage for 14 d, and Viagra was used as the positive control drug. Sexual behaviour was observed, and concentrations of serum testosterone, nitric oxide (NO), and cyclic guanosine monophosphate (cGMP) in serum as well as in penis were measured. In addition, weights of genital organs were also measured. RESULTS: The combined extracts of maca root and Chinese chive seed (1:1, w/w) had a 45-fold increase in macamide content compared with maca extract. It also led to significantly higher ejaculation frequency (P < 0.05) than single extract from maca root or Chinese chive seed, with no corresponding effect on genital indices. In addition, the NO level in serum (P < 0.01) and penis (P < 0.05) increased notably, as well as the level of cGMP in penis (P < 0.05). CONCLUSIONS: The results indicated that the combined extracts produced better synergistic effects on male sexual function than maca extract or Chinese chive extract alone. These positive effects may involve the upregulation of NO and cGMP concentrations in penis.

Homogenate-assisted high-pressure disruption extraction for determination of phenolic acids in Lonicerae Japonicae Flos.

An effective method based on the combined homogenate-assisted high-pressure disruption extraction (HHPDE) was applied to the extraction and determination of the main phenolic acid compounds from Lonicerae Japonicae Flos. The optimized HHPDE showed competitive advantage in yield (The extraction yields of NCA, CA, 3,5DCA and 4,5DCA in HHPDE were 1.21, 1.08, 1.06 and 1.17 fold higher than those in UAE), time-saving (<5 min) and relative low temperature requirement (4-16 °C) compared to HRE and UAE. Furthermore, the HHPDE method behaved a good repeatability and reproducibility according to the HPLC. The mentioned HHPDE method is firstly applied in the extraction and quantification of neochlorogenic acid chlorogenic acid, 3,5-dicaffeoylquinic acid and 4,5-dicaffeoylquinic acid in Lonicerae Japonicae Flos. This work provided an excellent alternative for the extraction and quantification of thermosensitive from plants.

Precise Dissolution Control and Bioavailability Evaluation for Insoluble Drug Berberine via a Polymeric Particle Prepared Using Supercritical CO2.

It is still controversial whether poor aqueous solubility is the most primary reason for the low oral bioavailability of insoluble drugs. Therefore, in this study, berberine-loaded solid polymeric particles (BPs) of varied dissolution profiles with ß-cyclodextrin (ß-CD) as carrier were fabricated using solution-enhanced dispersion by supercritical fluids (SEDS), and the relationship between dissolution and berberine (BBR) bioavailability was evaluated. Dissolution property was controlled via particle morphology manipulation, which was achieved by adjusting several key operating parameters during the SEDS process. Characterization on BP using infrared spectroscopy, differential scanning calorimetry, and X-ray diffraction indicated that BBR was dispersed in amorphous form, while nuclear magnetic resonance spectroscopy showed that methoxy groups of BBR were included into the cavities of ß-CD. In vivo pharmacokinetic studies showed that oral bioavailability increased by about 54% and 86% when the dissolution rate of BBR was increased by 51% and 83%, respectively. The entry speed of BBR into the bloodstream was also advanced with the degree of dissolution enhancement. It seemed that dissolution enhancement gave positive effect to the oral bioavailability of berberine, but this might not be the crucial point. Meanwhile, supercritical CO2 technology is a promising method for pharmaceutical research due to its advantages in regulating drug-dosage properties.

Qualitative and Quantitative Characterization of Monosaccharide Components of Salvia miltiorrhiza, Liguspyragine Hydrochloride, and Glucose Injection.

Salvia miltiorrhiza, liguspyragine hydrochloride, and glucose injection (SLGI) was made of Salvia miltiorrhiza Bge., liguspyragine hydrochloride, glucose, and glycerin. There were many kinds of monosaccharide components in SLGI, which might be from the raw material and Salvia miltiorrhiza Bge. Separation was performed on a Phenomenex Luna C18 analytical column (250 mm × 4.6 mm i.d., 5 µm, AccuStandard Inc., USA) at 30°C. The mobile phase consisted of two solvents: 0.1 mol/L phosphate-buffered saline (pH 6.7) (solvent A) and acetonitrile (solvent B) with gradient elution. The flow rate was maintained at 1.0 mL/min. Five kinds of monosaccharide components, glucose, D-mannose, L-rhamnose monohydrate, galactose, and xylose, were detected by precolumn derivatization HPLC, and their contents were compared with each other. And finally, concentrations of glucose in SLGI were determined and they were higher than the values of marked amount, which showed that one source of glucose might be from Salvia miltiorrhiza Bge. in SLGI. The average concentration of glucose was 5.18 g/100 mL, which was near the average value at 5.25 g/100 mL detected by ultraviolet spectrophotometry and also close to the marked amount (5.00 g/100 mL) on the label.

Ganoderin A, a novel 9,11-secosterol from Ganoderma lucidum spores oil.

In this study, four sterols were isolated from the Ganoderma lucidum spores oil obtained via supercritical CO2 extraction. Four chemical constituents were ganoderin A (1), chaxine B (2), ergosterol, (3) and stellasterol (4). All the separated ingredients were characterized using spectral data interpretation and by comparing with reported data. Noticeably, stellasterol and chaxine B were both firstly isolated from Ganoderma lucidum spores oil and ganoderin A was shown to bear an unprecedented skeleton.

Near-Infrared Spectroscopy Assay of Key Quality-Indicative Ingredients of Tongkang Tablets.

The objective of this paper is to develop an easy and fast near-infrared spectroscopy (NIRS) assay for the four key quality-indicative active ingredients of Tongkang tablets by comparing the true content of the active ingredients measured by high performance liquid chromatography (HPLC) and the NIRS data. The HPLC values for the active ingredients content of Cimicifuga glycoside, calycosin glucoside, 5-O-methylvisamminol and hesperidin in Tongkang tablets were set as reference values. The NIRS raw spectra of Tongkang tablets were processed using first-order convolution method. The iterative optimization method was chosen to optimize the band for Cimicifuga glycoside and 5-O-methylvisamminol, and correlation coefficient method was used to determine the optimal band of calycosin glucoside and hesperidin. A near-infrared quantitative calibration model was established for each quality-indicative ingredient by partial least-squares method on the basis of the contents detected by HPLC and the obtained NIRS spectra. The correlation coefficient R 2 values of the four models of Cimicifuga glycoside, calycosin glucoside, 5-O-methylvisamminol and hesperidin were 0.9025, 0.8582, 0.9250, and 0.9325, respectively. It was demonstrated that the accuracy of the validation values was approximately 90% by comparison of the predicted results from NIRS models and the HPLC true values, which suggested that NIRS assay was successfully established and validated. It was expected that the quantitative analysis models of the four indicative ingredients could be used to rapidly perform quality control in industrial production of Tongkang tablets.

[Extraction of 10-Deacetyl Baccatin by Supercritical CO2 from Taxus yunnanensis Branches and Leaves].

OBJECTIVE: To study the supercritical CO2 fluids extraction (SFE) method to extract the components from Taxus yunnanensis. METHODS: Medicinal meterials were extracted by supercritical CO2, and then purified by industrial chromatography. Using the extraction yield of 10-DAB as the index,single factor test was carried out to investigate the effect of co-solvent, extraction time, extraction pressure, extraction temperature, pressure and temperature of separation kettle I. Then orthogonal experiment was used to optimize the best extraction condition. RESULTS: The suitable extraction condition was as follows: the ratio of co-solvent (80% ethanol) amount and the madicinal materials was 3: 1, Separation kettle I pressure was 14 MPa, separation kettle I temperature was 40 °C, extraction pressure was 25 MPa, extraction temperature was 60 T and extraction time was 90 min. The extract was separated by industrial chromatographic and then crystallized. CONCLUSION: The supercritical CO2 extraction and purification process of 10-DAB were simple and feasible.

[Fingerprint and Simultaneous Determination of Multi-components in Water-soluble Components of Salvia miltiorrhiza in Miao Autonomous County of Songtao, Guizhou].

OBJECTIVE: To establish the HPLC fingerprint of water-soluble components of Salvia miltiorrhiza in Songtao, Guizhou, and to perform simultaneous determination of six components in it, so as to provide analytical method for its quality control. METHODS: The analyses were performed on a Phenomenex Luna C18 (2) (250 mm x 4. 6 mm, 5µm) column eluted with 0. 4% formic acid(A) - acetonitrile(B) in a gradient mode. The flow rate was 1. 0 mL/min, column temperature was set at 30 °C. RESULTS: Eleven common peaks were identified form the HPLC fingerprint of Salvia miltiorrhiza from 10 batches, the HPLC fingerprint similarities of 10 batches were not less than 0. 999. The linear ranges of danshensu, protocatechuic aldehyde, caffeic acid, rosmarinic acid, lithospermic acid and salvianolic acid B were 0. 0680 ~ 1. 3583 mg/mL, 0. 0008 ~ 0. 3967 mg/mL, 0. 0005 ~ 0. 2660 mg/mL, 0. 0020 ~ 0. 3992 mg/mL, 0. 0063 ~ 0. 6311 mg/mL and 0. 0097 ~ 1. 9306 mg/mL with r ≥ 0. 9999, respectively. The recovery rates were 100. 84%,102. 44%, 100. 53% ,100. 63%, 100. 83% and 100. 35% with RSD <2. 3%, respectively. CONCLUSION: The established method is simple, accurate and can provide reference for quality control of Salvia miltiorrhiza.

[Study on Chemical Constituents of Fat-soluble Extraction from Lepidium meyenii].

OBJECTIVE: To study the chemical constituents of the fat-soluble extraction from Lepidium meyenii root. METHODS: Different extraction methods were studied, including supercritical carbon dioxide extraction, circumfluence extraction and steam distillation. Chemical constituents of the fat-soluble extraction from Lepidium meyenii were analyzed by GC/MS. RESULT: The number of compounds isolated by the above four methods were 38, 31, 14, 21 (specific gravity less than 1 in steam distillation) , and 25 (specific gravity greater than 1 in steam distillation), accounting for 85.79%, 81.18%, 62.08%, 98.36% (specific gravity less than 1 in steam distillation) and 81.54% (specific gravity greater than 1 in steam distillation) of each total peak area, respectively. CONCLUSION: This study lays a certain foundation for further study and development of functional factors in Lepidium meyenii root.

[Chemical components from essential oil of Pandanus amaryllifolius leaves].

OBJECTIVE: To analyze the chemical compositions of Pandanus amaryllifolius leaves essential oil extracted by steam distillation. METHODS: The essential oil of Pandanus amaryllifolius leaves was analyzed by gas chromatography-mass spectrum, and the relative content of each component was determined by area normalization method. RESULTS: 128 peaks were separated and 95 compounds were identified, which weighed 97.75%. The main chemical components of the essential oil were phytol (42.15%), squalene (16.81%), what's more pentadecanal (6.17%), pentadecanoic acid (4.49%), 3, 7, 11, 15-tetramethyl-2-hexadecen-1-ol (3.83%), phytone (2.05%) and the other 74 chemical compositions were firstly identified from the essential oil of Pandanus amaryllifolius leaves. CONCLUSION: The chemical compositions of Pandanu samaryllifolius leaves essential oil was systematically, deeply isolated and identified for the first time. This experiment has provided scientific foundation for further utilization of Pandanus amaryllifolius leaves.

[Supercritical CO2 extraction and component analysis of Aesculus wilsonii seed oil].

OBJECTIVE: To research the optimal extraction process of supercritical CO2 extraction and analyze the component of the oil extracted from Aesculus wilsonii seed. METHODS: Using the yield of Aesculus wilsonii seed oil as the index, optimized supercritical CO2 extraction parameter by orthogonal experiment methodology and analysed the compounds of Aesculus wilsonii seed oil by GC-MS. RESULTS: The optimal parameters of the supercritical CO2 extraction of the oil extracted from Aesculus wilsoniit seed were determined: the extraction pressure was 28 MPa and the temperature was 38 degrees C, the separation I pressure was 12 MPa and the temperature was 40 degrees C, the separation II pressure was 5 MPa and the temperature was 40 degrees C, the extraction time was 110 min. The average extraction rate of Aesculus wilsonii seed oil was 1.264%. 26 kinds of compounds were identified by GC-MS in Aesculus wilsonii seed oil extracted by supercritical CO2. The main components were fatty acids. CONCLUSION: Comparing with the petroleum ether extraction, the supercritical CO2 extraction has higher extraction rate, shorter extraction time, more clarity oil. The kinds of fatty acids with high amounts in Aesculus wilsonii seed oil is identical in general, the kinds of fatty acids with low amounts in Aesculus wilsonii seed oil have differences.

[Study on the immunocompetence of polysaccharide extracted from root of Salvia miltiorrhiza].

OBJECTIVE: To study the immune activity of polysaccharide extracted from root of Salvia miltiorrhiza. METHODS: Investigated the effects of polysaccharide extracted from root of Salvia miltiorrhiza on lymphocyte proliferation response of mouse induced by LPS (the lipopolysaccharide LPS), phagocytosis of the peritoneal macrophage of mice to chick erythrocytes and the mouse models of delayed type hypersensitivity (DTH) response induced by DNFB. RESULTS: Lymphocyte proliferation and phagocytosis of the peritoneal macrophage of mice could be promoted by the polysaccharide, which could inhibit ear edema and capillary permeability increase induced by DNFB and enlarged the thymus and splenic index in mice. The expression of iNOS, IFN-alpha and IL-1beta was inhibited significantly in the treatment group. CONCLUSION: The polysaccharide extracted from Salvia miltiorrhiza can improve immune function.