Background: Hypericum elegans is used in Bulgarian folk medicine for treatment of wounds, depression, gastrointestinal and bacterial diseases. Objective: Recently, new natural benzophenones: Elegaphenone and O-glycosides: Hypericophenonoside, Neoannulatophenonoside and Elegaphenonoside as well as already known 7-Epiclusianone were isolated from the titled species. The aim of the present study was to evaluate the antioxidant and acetyl cholinesterase inhibitory potential of the isolated compounds. Materials and Methods: 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) di-ammonium salt (ABTS) free radicals, ferric reducing antioxidant power (FRAP) assay as well as inhibition of lipid peroxidation in linoleic acid system were used for determination of antioxidant activity. Modified Ellman's colorimetric method was carried out to assess the acetyl cholinesterase inhibition potential. Hyperoside and Galantamine hydrobromide were used as positive controls. Results: Hypericophenonoside was found to possess the strongest DPPH radical scavenging activity (IC ₅₀ = 181.85 ± 6.82 μM), while Neoannulatophenonoside showed the highest ABTS (IC ₅₀ = 0.25 ± 0.005 μM) and lipid peroxidation inhibitor activity. FRAP activity was demonstrated only by prenylated aglycones - Elegaphenone [942.16 ± 4.03 μM Trolox Equivalent (TE)] and 7-Epiclusianone (642.95 ± 3.95 μM TE) and was stronger compared to the control Hyperoside (421.75 ± 9.29 μM TE). Elegaphenone and 7-Epiclusianone were found to possess moderate acetyl cholinesterase inhibitory potential with IC ₅₀ values of 192.19 ± 3.54 μM and 142.97 ± 4.62 μM, respectively. Conclusion: The results obtained revealed that H. elegans is a potential natural source of bioactive compounds and benzophenones could be useful in therapy of free radical pathologies and neurodegenerative disorders.

Purpose: To formulate and optimize a herbal gel of Aloe vera extract containing Carbopol 934 as gelling agent and to investigate the effects of topical application of Carbopol 934 gel containing Aloe vera extract on the healing of skin wounds surgically induced in Wistar rats. Materials and Methods: Different concentrations of viscosity enhancer Carbopol 934 were tried and finally gel that showed good spreadability and consistency was selected for wound healing property of herbal gel of Aloe vera. Excision wound model was used for the study. Results: The optimized gel was evaluated for different physicochemical properties and wound healing property. Differences in wound healing were observed between the various treatments when compared to the control group. Tissue hyperplasia was lower in the control group compared to the other treated groups. In animals group treated with gel, 80.14% healing was observed up to 14 ^th day. While in untreated group I (control) animals showed 52.68% healing of wounds on 14 ^th day. On the other hand, control group animals also showed inflammation and pus formation up to 5 ^th day of study, while treated animals did not showed any observable inflammation and pus formation. Conclusion: Results shows prepared gel has promising effect on the wound healing process.

Background: The content of icaritin and genistein in herba is very low, preparation with relatively large quantities is an important issue for extensive pharmacological studies. Objective: This study focuses on preparing and enzymic hydrolysis of flavonoid glycosides /β-cyclodextrin inclusion complex to increase the hydrolysis rate. Materials and Methods: The physical property of newly prepared inclusion complex was tested by differential scanning calorimetry (DSC). The conditions of enzymatic hydrolysis were optimized for the bioconversion of flavonoid glycosides /β-cyclodextrin inclusion complex by mono-factor experimental design. The experiments are using the icariin and genistein as the model drugs. Results: The solubility of icariin and genistein were increased almost 17 times from 29.2 μg/ml to 513.5 μg/ml at 60^˚ C and 28 times from 7.78 μg/ml to 221.46 μg/ml at 50^˚ C, respectively, demonstrating that the inclusion complex could significantly increase the solubility of flavonoid glycosides. Under the optimal conditions, the reaction time of icariin and genistin decreased by 68% and 145%, when compared with that without β-CD inclusion. By using this enzymatic condition, 473 mg icaritin (with the purity of 99.34%) and 567 mg genistein(with the purity of 99.46%), which was finally determined by melt point, ESI-MS, UV, IR, ¹ H NMR and ¹³ C NMR, was obtained eventually by transforming the inclusion complex(contains 1.0 g substrates). Conclusion: This study can clearly indicate a new attempt to improve the speed of enzyme-hydrolysis of poorly water-soluble flavonoid glycosides and find a more superior condition which is used to prepare icaritin and genistein.

Background: The RP-HPLC resolution of two triterpenic acid isomers was unstable. Objective: To separate the oleanolic acid (OA) and ursolic acid (UA) simply within RP-HPLC. Materials and Methods: The separation ability of five stationary phases was studied with the retention effect of their carbon loads. Also the resolution effects of mobile phase composition and different column temperatures were systematically investigated by using Drylab; (Rheodyne LLC.) after evaluating chromatograms automatically. Results: The best available resolution of two bioactive isomers was achieved (r = 3.4) via using PAH (polycyclic aromatic hydrocarbons) polymeric C18 bonded phase column. The chromatographic system was applied to the quantification in ten Chinese medicinal plants and the validation was carried out and the precision (RSD ≤1.34%), the linearity (r ≥0.9998) and the recovery (range from 92.1% to 102.6%) were acceptable. Conclusion: It is clear that the method was simple, rapid and reliable for the quantification of two compounds in new HPLC method within PAH polymeric C18.

Background: Picria fel-terrae is a traditional Chinese medicine. Materials and Methods: A new approach to the search for acetylcholinesterase (AChE) inhibitors from Picria fel-terrae is presented. Results: Bioassay- and LC-MS-guided fractionation of the ethyl acetate extract was from traditional Chinese medicine P.fel-terrae. Following primary extraction, the ethyl acetate extracts fraction of P.fel-terrae showed strong AChE inhibitory activities. So the sample was separated using highperformance liquid chromatography (HPLC). The effluent was split towards two identical 96-well fraction collectors, and the presence of the biologically interesting portion and chromatographic fractions could be readily detected by analyzing selected ion chromatograms through an electrophoresis-electrospray ionization mass spectrometry (ESIMS) system for accurate mass measurement. One 96-well plate was used for a bioassay (AChE-inhibitory assay) and detected the bioactivity and position of the relevant peak in the chromatogram. The positive well in the second 96-well plate was used for identification by LC-(+) ESIMS. Conclusion: As abovementioned, the AChE inhibitory constituents from P.fel-terrae by LC-bioassay-ESIMS were rapid identified. Liquid chromatography/ mass spectrometry (LC-MS) screening detected the presence of six active compounds, identified as picfeltarraenin IA (1), picfeltarraenin IB (2), picfeltarraenin IV (3), picfeltarraenin X (4), picfeltarraenin XI (5), and one unknown compound. The structures were further determined by 13C NMR. The six compounds expressed stronger AChE inhibition than the known AChE inhibitorTacrine. Above all, the value of this LC-bioassay-ESIMS methodology is highlighted by the finding and structure elucidation of the active constituents from many other structural families of natural products.

Background: Sturgeon (Acipenser sturio Linnaeus) skin contains high amount of nutrients including unsaturated fatty acids and collagen. A pepsin-assisted extraction procedure was developed and optimized for the extraction of collagen from Chinese sturgeon (Acipenser sturio Linnaeus) skins. Objective: To determine the optimum conditions with the maximum yield of the pepsin-soluble collagen (PSC) extraction. Materials and Methods: The conditions of the extraction were optimized using response surface methodology. The Box-Behnken design was used to evaluate the effects of the three independent variables (extraction time, enzyme concentration, and solid-liquid ratio) on the PSC yield of the sturgeon skin. Results: The optimal conditions were: solid-liquid ratio of 1:11.88, enzyme concentration of 2.42%, and extraction time of 6.45 h. The maximum yield of 86.69% of PSC was obtained under the optimal conditions. This value was not significantly different from the predicted value (87.4%) of the RSM (P < 0.05). Conclusion: The results of this study indicated that the production of PSC from sturgeon skin is feasible and beneficial. The patterns of sodium dodecyl sulfate-polyacrylamide gel electrophoretic patterns (SDS-PAGE) indicated that the sturgeon skin contains type I collagen, which is made of α-chain and β-chain. The infrared spectra of the collagens also indicated that pepsin hydrolysis does not affect the secondary structure of collagen, especially triple-helical structure.

Background: Ginseng cultivated under mountainous forest, called "Lin-Xia-Shan-Shen" (LXSS) in China's Pharmacopoeia. In recent years, it has been quickly propelled to plant at a large scale. Objective: To study the profiles of ginsenosides and carbohydrate profiles of LXSS. Materials and Methods: The contents of ginsenosides and carbohydrates, such as soluble sugar, polysaccharide, pectin, and starch in LXSS, were determined. All the above components were profiled, and the correlations between them were analyzed. Results: The results indicated that the contents of total ginsenoside, protopanaxadiol, protopanaxatriol, Rg ₁ , Re, Rb ₁ , Rc, Rb ₂ , Rd, starch, and pectin were negatively correlated with the growing years within 17 years. Among them, the content of starch was positively correlated with that of pectin. The total ginsenosides was positively correlated with starch and pectin, which cannot be found in garden ginseng, maybe resulting of fertilizer and other manual intervention in process of cultivation of garden ginseng. Discussion and Conclusions: The accumulation of ginsenosides and carbohydrate, especially starch and pectin, was different in garden ginseng and LXSS. This research may provide the scientific basis for germplasm evaluation, the cultivation and utilization of ginseng cultivated under mountainous forest.

Background: Clinically, significant herb-drug interactions have been previously documented and can be pharmacodynamic and/or pharmacokinetic in nature. Pharmacokinetic interactions have been attributed to induction or inhibition of either metabolic enzymes or efflux transporters. Objective: The effect of gel and whole leaf materials from 3 different aloe species namely Aloe ferox, Aloe marlothii, and Aloe vera as well as polysaccharides precipitated from the A. vera materials on the bi-directional transport of cimetidine across rat intestinal tissue was investigated. Materials and Methods: Cimetidine transport studies were performed across excised rat intestinal tissue mounted in Sweetana-Grass diffusion chambers in both the apical-to-basolateral and basolateral-to-apical directions. Results: While A. vera gel and whole leaf materials did not inhibit the efflux of cimetidine, the polysaccharides precipitated from them did show a reduction of cimetidine efflux. On the other hand, both A. ferox and A. marlothii gel and whole leaf materials exhibited an inhibition effect on cimetidine efflux. Conclusions: This study identified a modulation effect of efflux transporters by certain aloe materials. This may cause herb-drug pharmacokinetic interactions when drugs that are substrates for these efflux transporters are taken simultaneously with aloe materials. On the other hand, these aloe materials may be used for drug absorption enhancement for drugs with low bioavailability due to extensive efflux.

Background: Natural products might alter the labeling of blood constituents with technetium-99m ( ^99m Tc) and these results may be correlated with modifications of the shape of the red blood cells (RBC). The biodistribution of radiopharmaceuticals can be also altered. Objective: This investigation aimed to determine biological effects of an aqueous extract of chamomile (CE). Materials and Methods: To study the effect of the CE on the labeling of blood constituents with ^99m Tc, in vitro and in vivo assays were performed. The effect of the CE on the morphology of RBC was observed under light microscope. The images were acquired, processed, and the perimeter/area ratio of the RBC determined. To analyze the effect of the CE on biodistribution of the sodium pertechnetate (Na ^99m TcO ₄ ) in Wistar rats, these animals were treated or not with a CE. Na ^99m TcO ₄ was injected, the rats were sacrificed, the organs were removed, weighted and percentage of radioactivity/gram calculated. Result: In the in vitro experiment, the radioactivity on blood cells compartment and on insoluble fractions of plasma was diminished. The shape and the perimeter/area ratio of the RBC were altered in in vitro assays. An increase of the percentage of radioactivity of Na ^99m TcO ₄ was observed in stomach after in vivo treatment. Conclusion: These results could be due to substances of the CE or by the products of the metabolism of this extract in the animal organism. These findings are examples of drug interaction with a radiopharmaceutical, which could lead to misdiagnosis in clinical practice with unexpected consequences.

Background: Psoralea corylifolia (Bakuchi), a weed, which possesses a highly potent and medicinally important compound psoralen. P. corylifolia has been widely exploited since ages for its biological potential. Materials and Methods: Fifteen root nodulating bacteria as pure culture collection (PCC) were isolated from P. corylifolia in India. Further, these strains were evaluated for their effect on the psoralen content in P. corylifolia. High performance liquid chromatography (HPLC) method was used for the estimation of psoralen in P. corylifolia seed extracts. The effectiveness of these rhizobial strains was assessed on the basis of screening of various plant growth promoting attributes. Results: The 16S ribosomal RNA sequencing analysis revealed the identity of two most effective rhizobial isolates PCC2 and PCC7 as Rhizobium leguminosarum and Sinorhizobium meliloti, respectively. The R. leguminosarum PCC2 (JN546144) and Ensifer meliloti PCC7 (JN546145) strains showed solubilization of insoluble inorganic phosphate, secreted indole acetic acid (IAA), produced siderophore, showed ACC deaminase activity, and were positive for nodulation and nitrogen fixing genes. Seeds of P. corylifolia were bacterized with combination of R. leguminosarum PCC2 and Ensifer meliloti PCC7 along with their individual application that resulted in enhancement of various early vegetative and late reproduction parameters of plants in two consecutive field trials in the year 2009 and 2010. The psoralen content in the seeds of P. corylifolia was observed to be increased in the field trials where the combination of rhizobial strains PCC2 and PCC7 was used (2.79%) compared to control (1.91%). Conclusion: These findings indicate that rhizobial strains PCC2 and PCC7 showing good plant growth promoting attributes can be effective for increasing the psoralen content in the seeds of P. corylifolia to a certain level.

Objective: Consumption of green-leafy vegetables is being advocated beneficial for type 2 diabetes mellitus individuals possibly because they are cost effective source of potent biological antioxidants. This research analyzed various phytochemicals, free radicals scavenging antioxidant potentials and starch digesting enzymes inhibitory activities in fresh juice of nine green-leafy vegetables. Furthermore, this study also investigated influence of these vegetables juice on starch and glucose induced postprandial glycemic load. Materials and Methods: Phytochemical constituents, in vitro free radicals scavenging antioxidant and enzymes inhibitory activities were evaluated applying various reported methods. Post-prandial glycemic excursion was induced in rats pretreated with vegetables juice by oral administration of starch and glucose. Results: All the leafy vegetables juice displayed potent free radicals scavenging activities. Juice of amaranthus, rumex, palak and raphanus displayed potential anti-oxidative property by reducing H ₂ O ₂ induced hemolysis in rats red blood cells RBCs. Ajwain and rumex juice showed pancreatic α-amylase inhibitory activity. Alternanthera, ajwain, methi, amaranthus and sowa leaves juice displayed intestinal α-glucosidase inhibitory activity. Juice of raphanus, ajwain and sowa significantly mitigated starch-induced postprandial glycemic load. Amaranthus leaves juice potently mitigated glucose-induced postprandial glycemic load and also reduced hemoglobin glycation induced by glucose in vitro. Conclusions: This investigation finds that juice of leafy vegetables is potent source of biological antioxidants. In addition, juice of raphanus, ajwain and sowa leaves possess capacity to mitigate starch induced postprandial glycemic burden and amaranthus leaves' juice can reduce glucose induced postprandial glycemic excursion.