A practical and rapid screening method for influenza virus neuraminidase inhibitors based on fluorescence detection.

A new analytical method for rapid screening of influenza virus neuraminidase inhibitors was established. The method is based on the principle that, given a certain amount of neuraminidase, the sample and the neuraminidase act in the microplate for a period of time, and the active neuraminidase that is not inhibited by the sample can generate a fluorescence value at a specific wavelength after binding to the substrate, and the rate of inhibition of neuraminidase by the sample can be calculated based on the actual detected fluorescence value. This newly developed method was used to screen and evaluate the in vitro anti-neuraminidase activity of 39 high-purity compounds contained in three traditional Chinese herbal medicines, and finally 25 compounds with strong activity were obtained. The newly established neuraminidase inhibitor analytical method has these advantages of practicality, rapidity, high sensitivity and low cost, and has a good value for promotion and application. This article newly establishes a rapid, sensitive, simple and practical screening method for influenza virus neuraminidase inhibitors, which is a great complement to the existing methods and has a good promotion and application value.

[Chemical analysis of classical prescription Qianghuo Shengshi Standard Decoction by UHPLC-Q Exactive Orbitrap MS].

A UHPLC-Q Exactive Orbitrap MS method was used to analyze the chemical constituents of the classical prescription Qianghuo Shengshi Standard Decoction(QHSS). UHPL conditions were as follows: Waters~(TM) UPLC~(TM) HSS T3 C_(18) column(2.1 mm×100 mm, 1.7 µm) and mobile phase of acetonitrile-0.1% formic acid aqueous solution. Mass spectrometry data of QHSS, each herb extract, and negative sample were collected in both positive and negative ion modes. The chemical constituents of QHSS were identified or tentatively identified based on the accurate molecular weight, retention time, MS fragmentation, comparison with reference substances, and literature reports. A total of 141 compounds were identified, including 18 amino acids, oligosaccharides, oligopeptides, and their derivatives, 19 phenolic acids, 44 coumarins, 18 flavonoids and chromones, 13 saponins, 17 phthalides, and 12 other components. This study comprehensively characterized the chemical constituents of QHSS, laying an experimental basis for the in-depth research on the material basis and quality control of QHSS.

Traditional Uses, Pharmacological Effects, and Molecular Mechanisms of Licorice in Potential Therapy of COVID-19.

The current Coronavirus disease 2019 (COVID-19) pandemic has become a global challenge, and although vaccines have been developed, it is expected that mild to moderate patients will control their symptoms, especially in developing countries. Licorice, not only a food additive, but also a common traditional Chinese herbal medicine, which has several pharmacological effects, such as anti-inflammation, detoxification, antibacterial, antitussive, and immunomodulatory effects, especially in respiratory diseases. Since the outbreak of COVID-19, glycyrrhizin, glycyrrhizin diamine and glycyrrhizin extract have been widely studied and used in COVID-19 clinical trials. Therefore, it is a very interesting topic to explore the material basis, pharmacological characteristics and molecular mechanism of licorice in adjuvant treatment of COVID-19. In this paper, the material basis of licorice for the prevention and treatment of COVID-19 is deeply analyzed, and there are significant differences among different components in different pharmacological mechanisms. Glycyrrhizin and glycyrrhetinic acid inhibit the synthesis of inflammatory factors and inflammatory mediators by blocking the binding of ACE 2 to virus spike protein, and exert antiviral and antibacterial effects. Immune cells are stimulated by multiple targets and pathways to interfere with the pathogenesis of COVID-19. Liquiritin can prevent and cure COVID-19 by simulating type I interferon. It is suggested that licorice can exert its therapeutic advantage through multi-components and multi-targets. To sum up, licorice has the potential to adjuvant prevent and treat COVID-19. It not only plays a significant role in anti-inflammation and anti-ACE-2, but also significantly improves the clinical symptoms of fever, dry cough and shortness of breath, suggesting that licorice is expected to be a candidate drug for adjuvant treatment of patients with early / mild COVID-19.

Synthesis, characterization and inhibitory effects of crocetin derivative compounds in cancer and inflammation.

Crocetin is a natural product possessing extraordinary therapeutic effects for various diseases. However, its extremely low solubility limits its application greatly. Conjugation of organic compounds containing heteroatoms such as N to poor soluble molecules can help the synthesized derivative to form stable hydrogen bonds by lowering the salvation energy, which will improve the solubility of the synthesized compounds. Herein, crocetin was modified by conjugating with piperidyl, diethylin and benzylamine to improve their solubility and bioactivities. In the present study, the conjugation of crocetin with piperidyl, diethylin and benzylamine and their influence on the solubility and the pharmacological effects of crocetin were investigated. With the described strategy, crocetin derivatives were synthesized and their structures were elucidated by 1H NMR, 13C NMR and UPLC-MS spectroscopic analysis. The solubility of crocetin and its derivatives were identified. Upon that, the pharmacological effects of the crocetin derivatives on the tumor and inflammation treatment were investigated. It was shown that, in contrast to crocetin, of which, the solubility and pharmacological effects were low and limited, the synthesized compounds have significantly higher solubility and possess broad spectrum of anticancer effects in multiple tumor cell lines, including B16F10, MCF-7, A549 and SKOV3, as well as enhanced anti-inflammation efficacy in macrophage (RAW264.7) without causing cells damage. Conjugation of piperidyl, diethylin and benzylamine with the crocetin was demonstrated to be a highly efficient strategy to improve the solubility of crocetin. The synthesized crocetin derivatives were shown the promising therapeutics for the tumor and inflammation treatment with high safety.

[Effect of nitrogen, phosphorus and potassium fertilizers on Flos Sophorae Immaturus yield and quality in hilly area of Chongqing].

With Sophora japonica at the flowering stage as the object, the effect of nitrogen, phosphorus and potassium fertilizers on the yield composition factors, yield and quality of Flos Sophorae Immaturus (FSI) was studied. The results indicated that in early spring, nitrogen, phosphorus and potassium fertilizer on the amplification rate of S. japonica, FSI yield composition, yield and quality were different significantly, middle to high nitrogen (1.5-2.0 kg/plant) significantly increased the level of panicled clusters, raceme and flower bud number and yield. Phosphorus (1.5-2.0 kg/plant) could significantly increase the total buds of flower number and yield, potassium showed no significant increase in yield and yield components. Comprehensively considering yield and quality of FSI, nitrogen 1.5-2.0 kg/plant, phosphorus 1.5-2.0 kg/plant and potassium 0.6-0.9 kg/plant are appropriate.

Synthesis and cardiomyocyte protection activity of crocetin diamide derivatives.

A series of novel diamide derivatives (2-8) of crocetin (1) were synthesized and evaluated for their cardioprotective activity in vitro. Using well-established model of hypoxia-induced injury in H9c2 cells, we investigated the effects of 9 compounds and positive drug nicorandil on cellular cytotoxicity by MTT assay, mitochondrial viable staining, LDH activity and mitochondrial membrane potential (MMP). Among the new derivatives, compounds 3 and 4 with good liposolubility showed significantly potent activity than crocetin (1) against hypoxia-induced cytotoxicity. Further mechanisms studies indicated that the cardioprotective effect of compounds 3 and 4 was due to these abilities by decreasing LDH release, preserving mitochondrial viabilities and reducing oxidative stress-induced depolarization of MMP. Our results demonstrated that compounds 3 and 4 as a new class of crocetin diamide derivatives could be developed as potential agents in our further drug development studies for ischemic heart disease.

[Optimization of extraction process of total amino acid from Bubali Cornu by orthogonal test].

OBJECTIVE: To optimize the extraction conditions of the total amino acid from Bubali Cornu. METHOD: An orthogonal test of L9 (3(4)) was designed to select optimum exaction conditions of the total amino acid. The influence of solvent concentration, the dosage of solvent and time of extraction were investigated with the content of total amino acid and thrombin-induced fibrin clotting time as index. RESULT: The optimum extraction condition procedure was described as follows: the concentration of the solvent was 4 mol x L(-1), volume of solvent was 6 times amount of the materials. CONCLUSION: The optimum exaction conditions procedure is reasonable and stable.