The rapid development of antibiotic resistance among bacterial pathogens has threatened to take humans back to the "pre-antibiotic era". The New Delhi metallo-β-lactamase (NDM-1) hydrolyzes nearly all β-lactam antibiotics including carbapenems. Bacterial strains carrying blaNDM-1 gene are termed "superbugs" and clinical inhibitors of NDM-1 have not yet been identified. Discovery of novel NDM-1 inhibitors is a challenging but rewarding research area. This review focuses on the structural characteristics and catalytic mechanisms of NDM-1, and comprehensively summarizes the development of NDM-1 inhibitors in order to facilitate the further development of NDM-1 inhibitors.

Although noninvasive trachea cannula is a common technique in clinic, this technique is hard to be applied in mice. It is even more difficult to perform the repeated noninvasive trachea cannula on the same mouse due to the throat injury. Here we report an effective, rapid and easy method to perform the noninvasive intubation on mice. This method can effectively be applied to many pharmacological and other related studies. In addition, this strategy can protect the mouse from physical injury caused by operative procedure. All procedures involving animal treatment were approved according to the Committee on the Ethics of Animal Experiments of the Institute of Materia Medica, Chinese Academy of Medical Sciences. In alliance with lung function measurement system, we detected the normal lung function values and presented the lung function development curves of mice of different age. We found that the lung function of mice was matured at 8 weeks old or at the body weight of 18-20 g. In addition, we constructed the mouse model of multiple bleomycin induced pulmonary fibrosis by using this method and illustrated that the trachea cannula is more efficiency than nasal inhalation. Using this method, we have confirmed that the immune microenvironment in lung tissue of multiple bleomycin model is distinct from that of single bleomycin induced pulmonary fibrosis model. Thus, this method is a reliable and safe strategy for the pharmacology study.

MicroRNAs (miRNAs) are a class of non-coding single-stranded RNAs involved in the regulation of gene expression found in a wide variety of eukaryotic cells and viruses. Recent studies have shown that some plant-derived miRNAs, which can stably exist in blood, tissues, and organs of animals, play a role in regulating the expression of different target proteins. In this review, we intend to sort out the mechanism of plant miRNA regulation based on the current research, and discuss its application prospects in the mining of miRNA active components of traditional Chinese medicine, small nucleic acid drug development, and drug development using plants as carriers. This can be beneficial to deepen the understanding of plant miRNA regulation, as well as the pharmacological mechanism and biological function of medicinal plants, thus providing new ideas for the prevention or treatment therapies towards human diseases.

This study aims to synthesize fluorinated hyperbranched poly(amido amine)s for the delivery of influenza DNA vaccine. Hyperbranched poly(amido amine)s (HP) were synthesized by using Michael-type polyaddition and then fluorinated to obtain fluorinated polymers (F-HP). The target gene was amplified by designing specific primers to construct the eukaryotic expression plasmid of influenza viral PR8 nucleoprotein (NP) gene as DNA vaccines. Then the polyplexes (F-HP/NP) were prepared by the electrostatic interactions between polymers and plasmid. The results suggested that the molecular weight of HP was 59.7 kDa and polydispersity index (PDI) was 2.67. The fluorine content in F-HP was found to be 20% (w/w). Transmission electron microscopy (TEM) revealed that the polyplexes had spherical shapes with sizes around 100 nm (w/w 5-10) and decreased with increasing w/w ratios. F-HP polyplexes showed improved cell uptake, lysosomal escape and elevated expression levels of NP in vitro than polyplexes based on HP. Finally, the in-vivo immunization by F-HP/NP polyplexes triggered improved CD8⁺ T cell responses. This study suggests that fluorinated hyperbranched poly (amido amine)s represent one of the effective carriers for DNA vaccine delivery. The animal experiments were approved by the Experimental Animal Ethics Committee of Yangzhou University.

In this study the blending process of Qingyan tablets was simplified and simulated, and near infrared spectroscopy was used to monitor the blending process of several raw materials in different particle size systems to explore the influence of particle size on the blending end-point. Five blended batches with different particle sizes were designed in this experiment and the near infrared spectra of the blended samples were collected. Partial least squares regression (PLSR) models of the contents of Platycodonis Radix, Fructus Chebulae, borax, Hanshuishi and microcrystalline cellulose were developed. A quantitative model was applied to determine the blending end-points of three separate batches of blends with different particle sizes. The moving block of standard deviation (MBSD) was used as a qualitative method for determination of the blending end-point. The results show that the smaller the particle size of the materials, the shorter the time to reach the blending end-points, and with more accurate model predictions. In addition, although the MBSD method is convenient and fast without modeling, the results of blending end-point determination were not as accurate as PLSR method. This approach allowed us to determine the blending end-point of Qingyan tablets.

Acid-sensing ion channel 1a (ASIC1a) is an ammonia-chlorine-sensitive ligand-gated ion channel, and is widely distributed and expressed in the central and peripheral nervous systems. In a physiological environment, cells maintain a stable pH value around 7.0-7.5 through various transport modes of H⁺. During the occurrence of some pathological conditions such as allergic asthma, nephritis, arthritis, enteritis, acute lung injury, and other inflammatory diseases, the anaerobic glycolysis of tissue produces H⁺ accumulation of lactic acid and ATP hydrolysis, resulting in tissue acidification and body fluids. The pH value drops sharply to around 4.0-6.0, which further activates ASIC1a, causing a sharp deterioration of the inflammatory disease. In recent years, targeting ASIC1a may be a potential treatment strategy. This review briefly summarizes the role of ASIC1a in inflammatory diseases and discusses the research progress of ASIC1a in inflammatory diseases.

The paper aims to study and compare the effects of Scutellaria baicalensis leaves (SLE) and Scutellaria baicalensis tea (STE) water extracts on the lifespan of Drosophila melanogaster, and explore their anti-aging mechanism through metabolomics. Lifespan, food intake and fertility were measured after administering different doses of SLE and STE to a Drosophila natural aging model, and the metabolic profile of Drosophila was analyzed by metabolomics and multivariate statistical methods. The results showed that SLE and STE (1, 3 g·L^-1) could significantly prolong the average lifespan, median life and maximum lifespan of male Drosophila, improve the fertility of Drosophila, and had no effect on the food intake. Through metabolomics technology, 14 differential metabolites related to aging were found, involving a total of five metabolic pathways. All differential metabolites were reversed after STE intervention, while 13 differential metabolites were reversed after SLE intervention. The results suggest that SLE and STE can delay aging of Drosophila, and the anti-aging mechanism is related to energy metabolism.

Huang-Kui-Si-Wu Formula (HKSWF) can reduce the accumulation of uremic toxin p-cresyl sulfate (PCS) and its precursor p-cresol (PC) in a rat model of chronic kidney disease (CKD) and delay the progression of CKD. However, the mechanism by which HKSWF decreases PC accumulation is not clear. This study investigated the effect of HKSWF on PC production in intestinal microbes as well as its mechanism of action. After CKD model rats were given HKSWF by intragastric administration, feces were collected to analyze the gut bacterial composition by 16S rDNA sequencing technology. All procedures were approved by the Institutional Animal Care and Use Committee of the Nanjing University of Chinese Medicine. The results showed that HKSWF inhibited PC production without decreasing the abundance of harmful bacteria. HPLC-UV-FLD was used to detect p-cresol. An in vitro anaerobic culture system was used to study the effect and mechanism of action of HKSWF on PC production in gut bacteria. The results show that food-derived tyrosine (Tyr) could significantly promote PC production in intestinal bacteria, and HKSWF (4000, 400, 40 μg·mL^-1) could dose-dependently inhibit PC production in gut bacteria in vitro. HKSWF inhibited bacterial PC synthesis by two pathways: it decreased the oxidation pathway from 82.83% to 38.87%, and increased the reductive pathway from 17.17% to 61.13%. This result suggests that HKSWF could inhibit PC production by switching tyrosine metabolism from an oxidative pathway to a reductive pathway. Secondly, HKSWF could directly inhibit the oxidative pathway of tyrosine and decrease the decomposition of PHA, thereby inhibiting PC production. These results suggest that HKSWF could inhibit the formation of harmful uremic toxins by modulating the metabolic pathway of PC in gut microbiota and thereby delaying CKD progression.

This study was performed to determine the metabolic profile of a new illicit drug, PX-2, in human liver microsomes. Q Exactive™ HF Quadrupole-Orbitrap LC-MS (LC-QE-HF-Orbitrap-MS) was employed to determine the metabolic sites and pathways of phase Ⅰ and phase Ⅱ metabolism. PX-2 was added to a microsomal incubation model to simulate human hepatic metabolism. The results showed that a total of 18 phase Ⅰ metabolites and 3 glucuronidated phase Ⅱ metabolites were generated, with the main metabolic pathways of phase Ⅰ metabolism including amide hydrolysis, fluoropentyl oxidative defluorination, benzyl hydroxylation, and carbazole ring hydroxylation. Based on the type and sites of metabolism, phase Ⅰ metabolites M1.1 (amide hydrolysis), M4.1 (carbazole cyclic hydroxylation), and M3.1 (oxidative defluorinative hydroxylation) are proposed to be potential poisoning markers. The results of this study provide a basis for identification of related drugs and establishment of testing methods in biological samples.

The injectable recombinant protein of exendin-4 and human serum albumin (HSA), E2HSA, is a long-acting glucagon like-peptide-1 (GLP-1) receptor agonist, which is in clinical research stage now. This study aimed to evaluate the protective effects of E2HSA on mouse islet β cell function in vitro and in vivo. In vitro, the mouse insulinoma cells NIT-1 were used to assay the effects of E2HSA on cell viability and proliferation. Besides, the water soluble cholesterol was adopted to induce cell injury, and then the effects of E2HSA on cell viability and apoptosis, as well as the mechanism, were studied. In vivo, the alloxan-induced hyperglycemic mice were repeatedly administered with E2HSA by subcutaneous injection, and the blood glucose, serum insulin, and content of insulin in islets were measured. All animal experiments were carried out with approval of the Experimental Animal Welfare Ethics Committee of the Institute of Materia Medica (Chinese Academy of Medical Sciences and Peking Union Medical College). The results showed that E2HSA significantly increased the viability of NIT-1 cells and the amount of bromodeoxyuridine (BrdU) incorporated in cells. Besides, the water soluble cholesterol significantly decreased the cell viability and induced apoptosis of NIT-1 cells, but E2HSA significantly reversed the injury of NIT-1 cells. Nevertheless, E2HSA significantly increased the expression of pancreatic duodenal homeobox-1 (PDX-1) and protein kinase B (PKB) of NIT-1 cells after injured by water soluble cholesterol. Furthermore, repeated injections of E2HSA significantly reduced the fasting and non-fasting blood glucose, increased the serum insulin level and raised the insulin content in β cells of alloxan-induced hyperglycemic mice. In conclusion, E2HSA could promote proliferation of NIT-1 cells, inhibit the water soluble cholesterol induced injury and apoptosis, and increase the insulin content in serum and islets of alloxan-induced hyperglycemic mice, suggesting the protective effects on pancreatic islet β cell function.