Asparagus cochinchinensis is a commonly used traditional Chinese medicine with steroidal saponins as its main active ingredients. Due to the structural similarity and size of the steroidal saponins, these compounds cannot always be effectively separated by a combination of normal phase silica gel column chromatography and reversed phase ODS column chromatography. In this experiment, chromatographic columns with different separation mechanisms were systematically screened, and it was found that a chiral chromatographic cellulose column could effectively separate these components. This column was used to separate 3 mixtures to obtain 6 single compounds (1-6). Structural identification showed that the singular structural difference between these poorly separated components resides in a terminal glycosyl group (xylose or rhamnose) in the C-3 glycosyl chain, and compounds 4 and 6 are two new steroidal saponins. Since the structures of compounds are often unknown during the isolation and purification of natural products, chiral columns are rarely used. This study suggests that chiral chromatographic columns are a valuable option for natural products that are difficult to separate by conventional means.

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.

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.

The adenosine triphosphate (ATP) liposome, prepared with the methods of film dispersion and ion-pairing was evaluated for its therapeutic effect on hypoxic brain damage. The appropriate formulation is adenosine disodium triphosphate, hexadecyl trimethyl ammonium bromide, soybean phospholipid, cholesterol with mass ratio of 1:1.98:8:3. The encapsulation efficiency of ATP liposome was (81.50 ±0.82) % and the loading efficiency was (6.79 ±0.07) %. In vitro release test and rheology test were conducted to investigate the physicochemical properties of ATP liposomes and empty gels respectively. The blank methylcellulose gel, followed with ATP liposome and ATP aqueous solution added to the methycellulose gel, were used for nasal administration in mice respectively. All experiments were approved by the Ethics Committee for Experimental Research in Academy of Military Medical Sciences. After 9 days of continuous administration, ATP liposome hydrogel increased the values of red blood cells and hemoglobin (P < 0.01) compared to ATP hydrogel and blank gel. And the ATP liposome hydrogel significantly increased the standard hypoxia tolerance time in mice compared to ATP hydrogel and blank gel after 13 days of nasal administration (P < 0.05). The immunohistochemical staining of mice hippocampus for the proapoptotic gene p53 showed that ATP liposome hydrogel was capable of protecting brain tissue in hypoxia. It is indicated that the prophylactic administration of ATP liposome nasal gel can significantly improve the hypoxia tolerance of mice, and the ATP liposome nasal gel was proved to be a promising anti-hypoxia preparation.

Cyclophosphamide (CPA) is the first-line chemotherapy for many tumors, but its overdose will lead to hepatotoxicity. This study aims to investigate whether the combined administration of oxymatrine (OMT) with CPA will aggravate the hepatotoxicity induced by CPA and its engaged mechanism. The expression of hepatic Cyp2b10 mRNA and CYP2B10 protein was detected by qPCR and Western blot in mice at different times after OMT (100 mg·kg^-1) administration. Mice were given with different doses of OMT (intragastric administration, ig) every day. At the same time, CPA (200 mg·kg^-1) was also intraperitoneally injected into mice every other day. After 10 days, serum alanine/aspartate aminotransferase (ALT/AST) activity, the mortality of mice and hepatic mRNA expression of Cyp2b10 were detected. Furthermore, the correlation among ALT/AST activity, the mortality and Cyp2b10 mRNA expression was analyzed. All animals were received humane care according to the institutional animal care guidelines approved by the Experimental Animal Ethical Committee of Shanghai University of Traditional Chinese Medicine. The results showed that OMT itself enhanced hepatic mRNA and protein expression of Cyp2b10 (P < 0.05), and increased liver enzymatic activity of CYP2B10 in mice (P < 0.05). In mice treated with CPA plus OMT, OMT obviously enhanced the mortality of mice induced by CPA (from 33.3% to 58.3%). The results of serum biochemical analysis and hepatic mRNA expression of Cyp2b10 showed that OMT further enhanced the increased serum ALT/AST activity and hepatic Cyp2b10 mRNA expression in mice (P < 0.05). There was a good correlation between serum ALT/AST activity and mortality or hepatic Cyp2b10 mRNA expression. These results showed that OMT could enhance hepatic Cyp2b10 mRNA expression and increase liver CYP2B10 enzymatic activity, and then promoted the metabolism of CPA, and thus aggravated CPA-induced hepatotoxicity in mice.

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.

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.

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.

Corona virus disease 2019 (COVID-19) is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. At present, there is no specific antiviral drug for this virus, and the main clinical treatment is support and symptomatic treatment. Direct targeting the virus and the host targets are two strategies for the development of antiviral drugs. At present, the research and development of COVID-19 therapeutic drugs has made some progress on both approaches. Here we review potential anti-SARS-CoV-2 drugs to discuss the antiviral mechanisms and potential of these drugs from the perspectives of virus and host. The role of traditional Chinese medicine in the treatment of COVID-19 is discussed along with the prospects for drug treatment strategies of COVID-19.

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.