Identification of Polar Constituents in the Decoction of Juglans mandshurica and in the Medicated Egg Prepared with the Decoction by HPLC-Q-TOF MS².

As a folk medicinal plant, Juglans mandshurica has been used for the treatment of cancer in China and Korea. Traditionally, J. mandshurica is decocted together with chicken eggs. Both the decoction and medicated eggs possess anti-tumor properties. Clarifying the constituents of the decoction and absorbed by the medicated eggs is essential for the investigation of the active principles of J. mandshurica. Herein, the medicated eggs were prepared by decocting raw chicken eggs, having unbroken shells, with the decoction of J. mandshurica. A systematic investigation of the chemical profile of the J. mandshurica decoction and the medicated egg extraction was conducted by HPLC-Q-TOF-MS². In total, 93 peaks, including 45 tannins, 14 naphthalene derivatives, 17 organic acids, 3 diarylheptanoids, 4 lignans, 3 anthraquinones, 1 flavonoid glycoside, 3 amino acids, and 3 nitrogenous compounds, were tentatively identified in the decoction. In the medicated egg extraction, 44 peaks including 11 organic acids, 3 amino acids, 3 nitrogenous compounds, 8 naphthalene derivatives, 3 diarylheptanoids, 15 tannins, and 1 lignan were tentatively identified. The chemical profile presented provided a detailed overview of the polar chemical constituents in J. mandshurica and useful information for the research of bioactive compounds of this plant.

Evaluation of the Nano-TiO2 as a Novel Deswelling Material.

Nano-TiO2 is widely applied in the automobile exhaust hose reels as a catalyst to reduce oxynitride emissions, including nitric oxide (NO). In the biomedicine field, NO plays an important role in vasodilation and edema formation in human bodies. However, the deswelling activity of nano-TiO2 has not been reported. Here, we demonstrated that nano-TiO2 can significantly degrade the production of NO in LPS-induced RAW264.7 mouse macrophages. Further study indicated that nano-TiO2 exhibited an effect on vascular permeability inhibition, and prevented carrageenan-induced footpad edema. Therefore, we prepared a nano-TiO2 ointment and observed similar deswelling effects. In conclusion, nano-TiO2 might act as a novel deswelling agent related with its degradation of NO, which will aid in our ability to design effective interventions for edema involved diseases.

Role of berberine in anti-bacterial as a high-affinity LPS antagonist binding to TLR4/MD-2 receptor.

BACKGROUND: Berberine is an isoquinoline alkaloid mainly extracted from Rhizoma Coptidis and has been shown to possess a potent inhibitory activity against bacterial. However, the role of berberine in anti-bacterial action has not been extensively studied. METHODS: The animal model was established to investigate the effects of berberine on bacterial and LPS infection. Docking analysis, Molecular dynamics simulations and Real-time RT-PCR analysis was adopted to investigate the molecular mechanism. RESULTS: Treatment with 40 mg/kg berberine significantly increased the survival rate of mice challenged with Salmonella typhimurium (LT2), but berberine show no effects in bacteriostasis. Further study indicated that treatment with 0.20 g/kg berberine markedly increased the survival rate of mice challenged with 2 EU/ml bacterial endotoxin (LPS) and postpone the death time of the dead mice. Moreover, pretreatment with 0.05 g/kg berberine significantly lower the increasing temperature of rabbits challenged with LPS. The studies of molecular mechanism demonstrated that Berberine was able to bind to the TLR4/MD-2 receptor, and presented higher affinity in comparison with LPS. Furthermore, berberine could significantly suppressed the increasing expression of NF-κB, IL-6, TNFα, and IFNß in the RAW264.7 challenged with LPS. CONCLUSION: Berberine can act as a LPS antagonist and block the LPS/TLR4 signaling from the sourse, resulting in the anti-bacterial action.

[Study on characteristic HPLC fingerprint of Polygonum orientale inflorescence].

OBJECTIVE: To establish a characteristic HPLC fingerprint of Polygonum orientale inflorescence, and to provide reference for its quality evaluation. METHODS: Taxifolin was used as reference. HPLC analysis was carried out with Diamonsil C18 column (200 mm x 4.6 mm, 5 microm) using acetonitrile -0.1% phosphoric acid(gradient elution)as mobile phase at flow rate of 1.0 mL/min. The detection wavelength was set at 280 nm and the column temperature was 30 degrees C. RESULTS: Eighteen common peaks were pointed out from the HPLC fingerprint of Polygonum orientale inflorescence from 12 different habitats. Among of them,four common peaks were identified as taxifolin, catechin, gallic acid and 3,3'-dimethyl ellagic acid-4-O-beta-D-glucoside. Analyzed by "Similarity Evaluation for Chromatographic Fingerprint of Traditional Chinese Medicine" software, the HPLC fingerprint similarities of 12 samples were more than 0.9. CONCLUSION: This method is repeatable and exclusive. It can be used for identification and quality control of Polygonum orientale inflorescence.

Corrigendum to "Role of Baicalin in Anti-Influenza Virus A as a Potent Inducer of IFN-Gamma".

[This corrects the article DOI: 10.1155/2015/263630.].

HPLC determination of malondialdehyde in ECV304 cell culture medium for measuring the antioxidant effect of vitexin-4''-O-glucoside.

To investigate the antioxidant effect of vitexin-4''-O-glucoside, a flavone glycoside, isolated from the leaves of Crataegus pinnatifida Bge. var. major, we developed a simple and sensitive high-performance liquid chromatography (HPLC) method to determine levels of malondialdehyde (MDA) in ECV304 cell culture medium after induction by tert-butyl-hydroperoxide (TBHP). The preparation of analyzed samples involved a one-step derivatization with thiobarbituric acid (TBA). HPLC analysis was performed on a Synergi Hydro-RP, a polar end-capped C18 column (250 x 4.6 mm, 4 mum), using an acetonitrile-ammonium acetate aqueous solution (10 mM, pH 6.8) as the mobile phase under linear gradient conditions with UV detection at 532 nm. The calibration curve was linear over 0.0125-1.25 microM MDA (r = 0.9951). Relative standard deviations (RSDs) of intra-day and inter-day precision were less than 6.1% and 5.0%, respectively. The mean recovery was 96.9 +/- 1.6%. The lower limit of quantification (LLOQ) of MDA was 0.0125 microM. This chromatographic method was successfully applied to investigating the in vitro antioxidant effect of vitexin-4''-O-glucoside. Vitexin-4''-O-glucoside (120 M) protected ECV304 cells from peroxidation induced by TBHP.

[Effect of baicalin on in vivo anti-virus].

OBJECTIVE: This study is about the in vivo antivirus effect of baicalin. METHOD: Adopting in vivo experiment, we administered one group of mice 2 days before infected with influenza virus (A/FM/1/47) , while administer the other group with baicalin one day after infected, to observe the effect of baicalin in protecting and curing mice against influenza virus. RESULT: Baicalin can obviously prolong the survival time of mice infected with influenza virus (A/FM/1/47) (P <0.01), eliminate the influenza virus in lung, reduce the hemagglutination tite and infectivity in pulmo, and reverse mice's pneumonic pathological changes. CONCLUSION: Baicalin can obviously counteract influenza virus.

[The extract of compound Radix Scutellariae on mRNA replication and IFN expression of influenza virus in mice].

OBJECTIVE: To study the extract of compound Radix Scutellariae on inhibiting the mRNA replication and inducing the INF expression of influenza virus in mice's vivo. METHODS: Influenza virus infected mice pneumono-adaption stock A/FM/1/47 (H1N1) evoked mice pneumonia taken as animal model. RT-PCR was adopted to measure the content of influenza virus's mRNA and INF in mice to study the effect of the extract on mRNA replication and IFN expression. RESULTS: The extract of compound Radix Scutellariae could inhibit influenza virus' s mRNA replication (P < 0.01) and induce interferon expression in mice (P > 0.05). CONCLUSION: The extract of compound Radix Scutellariae can induce interferon and inhibit influenza virus replication in mice.

Role of Berberine in the Treatment of Methicillin-Resistant Staphylococcus aureus Infections.

Berberine is an isoquinoline alkaloid widely used in the treatment of microbial infections. Recent studies have shown that berberine can enhance the inhibitory efficacy of antibiotics against clinical multi-drug resistant isolates of methicillin-resistant Staphylococcus aureus (MRSA). However, the underlying mechanisms are poorly understood. Here, we demonstrated that sub-minimum inhibitory concentrations (MICs) of berberine exhibited no bactericidal activity against MRSA, but affected MRSA biofilm development in a dose dependent manner within the concentration ranging from 1 to 64 µg/mL. Further study indicated that berberine inhibited MRSA amyloid fibrils formation, which consist of phenol-soluble modulins (PSMs). Molecular dynamics simulation revealed that berberine could bind with the phenyl ring of Phe19 in PSMα2 through hydrophobic interaction. Collectively, berberine can inhibit MRSA biofilm formation via affecting PSMs' aggregation into amyloid fibrils, and thereby enhance bactericidal activity of antibiotics. These findings will provide new insights into the multiple pharmacological properties of berberine in the treatment of microbial-generated amyloid involved diseases.

Role of Baicalin in Anti-Influenza Virus A as a Potent Inducer of IFN-Gamma.

Baicalin (BA) is a flavonoid compound purified from Scutellaria baicalensis Georgi and has been shown to possess a potent inhibitory activity against viruses. However, the role of BA in anti-influenza virus has not been extensively studied, and the immunological mechanism of BA in antiviral activity remains unknown. Here, we observed that BA could protect mice from infection by influenza virus A/PR/8/34 (H1N1), associated with increasing IFN-γ production, but presented no effects in IFN-γ or IFN-γ receptor deficient mice. Further study indicated that BA could inhibit A/PR/8/34 replication through IFN-γ in human PBMC. Moreover, BA can directly induce IFN-γ production in human CD4(+) and CD8(+) T cells and NK cells, and activate JAK/STAT-1 signaling pathway. Collectively, BA exhibited anti-influenza virus A (H1N1) activity in vitro and in vivo as a potent inducer of IFN-γ in major IFN-γ producing cells.