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ObjectiveTo study the virulence and biofilm inhibition effect of Fufang Huangbai Fluid Paint （FFHBFP） on methicillin-resistant Staphylococcus aureus （MRSA）， and to explore the antibacterial effect of FFHBFP on MRSA， which provides a theoretical basis and reference for clinical medication. MethodFirstly， the microdilution method and time–growth curve were used to determine the minimum inhibitory concentration （MIC） of FFHBFP and vancomycin （VAN） against MRSA and the effect on bacterial growth. The effects of FFHBFP and VAN on the inhibition of MRSA virulence factor lipase and restoration of hydrogen peroxide （H2O2） sensitivity were detected under sub-minimum inhibitory concentration （sub-MIC）. The inhibitory effect of FFHBFP and VAN on MRSA biofilm formation and maturation was detected by the microplate method. The morphological changes of mature biofilms before and after administration were observed under a scanning electron microscope （SEM）. Real-time polymerase chain reaction （Real-time PCR） was utilized to detect the effect of 50.600 g·L-1 concentration of FFHBFP on the expression of MRSA virulence gene crtM and biofilm-forming genes fnbA and icaA. Finally， molecular docking technology was used to predict the mechanism of potential antibacterial active ingredients of FFHBFP in inhibiting the virulence and biofilm of MRSA. ResultThe MIC of VAN was 2 mg·L-1， and VAN below 1 mg·L-1 exerted no effect on MRSA growth. The MIC of FFHBFP was not determined， while the 101.200-202.400 g·L-1 original solution inhibited MRSA growth. Compared with the blank group and the VAN group， sub-MIC （25.300-50.600 g·L-1 original solution） inhibited lipase and recovered MRSA sensitivity to H2O2 （P<0.01）. The results of the microplate method showed that FFHBFP （25.300-202.400 g·L-1 original solution） inhibited biofilm formation and maturation （P<0.05， P<0.01）. The SEM exhibited that FFHBFP made the structure of biofilm loose and the size of the bacteria varied. FFHBFP at 50.600 g·L-1 concentration can inhibit the expression of related virulence genes and biofilm-forming genes （P<0.05， P<0.01）， and molecular docking results also showed that the main antibacterial active ingredients in FFHBFP have good binding ability to the target. ConclusionFFHBFP that cannot directly kill MRSA exerts clinical efficacy by impairing virulence expression， biofilm formation， and other pathogenic properties.
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Objective:To evaluate the scientificity and feasibility of processing of Pinelliae Rhizoma by hot water washing （Tangxi）， and to provide reference for the development of related famous classical formulas. Method:Processing method of Pinelliae Rhizoma washed by hot water was established based on ancient Tangxi processing method， and the process conditions were optimized by single factor tests. The weight， moisture， ash， extract， total acid （calculated by succinic acid） contents and high performance liquid chromatography （HPLC） fingerprint of Pinelliae Rhizoma were compared before and after processing. In addition， the rabbit eye irritation test was conducted to evaluate the toxicity changes. Result:The processing method of Pinelliae Rhizoma washed by hot water was as following：washed by 4 times the amount of hot water at 80 ℃ for 10 times until clear water， transfused cross-section after incision， no or slight numbness in the mouth. The average moisture， ash， extract contents of Pinelliae Rhizoma washed by hot water were 9.34%， 1.71% and 4.22%， respectively. After being processed， the decline rates of weight and total acid content of Pinelliae Rhizoma were 7.49% and 43.31%. The HPLC fingerprint of Pinelliae Rhizoma before and after washing showed a decrease in all components， but there was no new chromatographic peak， and peak 9 （adenosine） reduced significantly. The results of rabbit eye irritation test showed that there was no obvious eye conjunctival irritation after washing， indicating that the toxicity of Pinelliae Rhizoma decreased obviously after washing. Conclusion:The established method of Pinelliae Rhizoma by Tangxi processing is stable and feasible， the aqueous extract of Pinelliae Rhizoma has no obvious eye conjunctival irritation after washing.
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Objective: To investigate the removal effect of 5 kinds of common flocculants on 6 kinds of toxic alkaloids in processing wastewater of Aconiti Radix. Method: HPLC was employed to determine contents of 6 kinds of alkaloids in the processing wastewater of Aconiti Radix before and after flocculation.The mobile phase consisted of acetonitrile-tetrahydrofuran(25:15)-0.1 mol·L-1 ammonium acetate solution(each 1 L contains 0.5 mL of glacial acetic acid) for gradient elution, volume flow rate was 1.0 mL·min-1, detection wavelength was 235 nm.The removal characteristics and effects of five common flocculants of ferric chloride, aluminum sulfate, polymeric ferric sulfate, polyaluminum chloride/polyacrylamide(PAC/PAM) and sodium polyacrylate were compared and preliminarily optimized. Result: The 5 kinds of flocculants had certain selectivity for different types of alkaloids, and the removal of 6 kinds of alkaloids showed obvious differences.Among them, the PAC/PAM attenuated relative superiority, when the pH 6, adding amount of 0.25 g·L-1 and PAC-PAM dosing ratio of 30:1, comprehensive removal effect was relatively good, removal rates of mesaconitine and hypaconitine was 85.4%and 58.3%, respectively. Conclusion: The flocculation method can be used as a pretreatment process to reduce the toxicity of processing wastewater of Aconiti Radix.
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Ischemic stroke is one of the diseases that seriously threaten human health. It is characterized by the high morbidity, disability rate and mortality, and has been lacking effective treatment. Its occurrence is related to metabolic disorder, calcium overload, free radical injury, inflammatory reaction, etc. Gardeniae Fructus not only has antipyretic, analgesic, hepatoprotective and cholagogic effects, but also has protective effects against ischemic brain injury. At present, there are more studies about the main components of Gardeniae Fructus against ischemic brain injury, but the mechanism is unclear. In this paper, the mechanism of the main active ingredients from Gardeniae Fructus in the treatment of cerebral ischemia injury in recent years was reviewed, and the effective component monomer and the whole were analyzed, so as to provide a basis for the prevention and treatment of ischemic brain injury of Gardeniae Fructus decoction pieces, and provide a reference for further research and application of this decoction pieces.
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Vascular dementia [VD] models were made first by repeating cerebral ischemia-reperfusion and followed by treating with estrogen. Learning-memory ability was measured by Morris water maze. Concentration of Ca[2+] in hippocampus was determined by Fura-2/AM fluorescence probe and the expression of Calbindin-D28K [CB] in hippocampal CA1 was tested by immunohistochemistry. Learning-memory ability was improved in E group rats; Concentration of Ca[2+] in hippocampus was decreased in E group rats. The expression of CB was less in E group rats. It implies that estrogen could improve learning-memory ability in VD rats, which may be associated with suppressing intracellular Ca[2+] overload and increasing the expression of CB in hippocampus
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To investigate the feasible application of the bioassay method in the evaluation of traditional Chinese medicine sustained-release preparations, develop a rapid drug-release evaluation method in vitro for multi-component preparations, and replace the biological activity determination method characterizing the overall behavior with the existing drug-release evaluation method for single component, in order to give better instruction for sustained-release preparations. HPLC was adopted to determine dissolution media, drug releasing rates, and accumulative releasing of active ingredients (salvianolic acid B, protocatechuic aldehyde and rosmarinic acid) of Salvia Miltiorrhiza hydrophilic gel matrix tablets. The ultraviolet spectroscopy was adopted to determine the antioxidant activity of release media, and evaluate the correlation between the drug-time curve of various drug components and the drug-time curve of the total antioxidant activity. The correlation coefficient between the drug-release curve of various components and the drug-time curve of the total antioxidant activity was higher than the critical value r 0.898 (P < 0.001). This indicated that the drug-release curve of the three phenolic acids and the drug-time curve of the total antioxidant activity had a good correlation in different conditions, such as dissolution media, release rates and component ratios. The bioassay method for determination was feasible, simple and convenient for preparation quality evaluation and prescription design in the place of in vitro dissolution.
Subject(s)
Antioxidants , Chemistry , Biological Assay , Methods , Delayed-Action Preparations , Chemistry , Drugs, Chinese Herbal , Chemistry , Kinetics , Salvia miltiorrhiza , Chemistry , Solubility , Tablets , ChemistryABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of different chromatographic conditions on QAMS relative correlation factors (RCF) and relative retention values (RT(R)).</p><p><b>METHOD</b>C18 columns were used with methyl alcohol-0.4% phosphoric acid water (85: 15) as the mobile phase. The detection wavelength was 254 nm, the column temperature was 30 degrees C, and the flow rate was 1.0 mL x min(-1). The five anthraquinones in Rhei Radix et Rhizoma were selected to be the objects of study. The RCF and RT(R) among aloe-emodin, rhein, chrysophanol, physcion and emodin were determined under different chromatographic conditions.</p><p><b>RESULT</b>Their RCFs showed no significant difference.</p><p><b>CONCLUSION</b>The RCFs among anthraquinones established by QAMS can be used as a constant in content determination of traditional Chinese medicines/patent traditional Chinese medicines.</p>