The α4β2-nicotinic acetylcholine receptor(nAChR) is a ligand-gated ion channel that is distributed throughout the nervous system. It is involved in the regulation of various neurotransmitters including acetylcholine, dopamine, γ-aminobutyric acid, and norepinephrine. α4β2-nAChR plays an important role in learning, memory, cognition, attention, inflammation, and pain. A large number of studies have shown that α4β2-nAChR is an important therapeutic target for neurological diseases such as Alzheimer's disease, Parkinson's disease, epilepsy, depression, nicotine dependence, pain, etc. It is an important target in the early diagnosis and curative effect detection of neurodegenerative diseases including Alzheimer's disease. This review summarizes the role, mechanisms and related drug research advances on α4β2-nAChR ligand drugs in neurological diseases, as well as providing a theoretical basis for identifying and developing more suitable α4β2-nAChR-related compounds.

Fibrosis is a pathological process of abnormal hyperplasia and excessive deposition of extracellular matrix during the process of repair after tissue and organ damage. Injury/inflammation caused by variously chronic diseases is a major trigger for fibrogenesis. Fibrosis of the liver and kidney is a common organ fibrosis. Recently, the intestinal microbiota has been shown to be extensively involved in the development of liver and kidney diseases, which may follow from changes in the intestinal microbial composition and intestinal integrity. This promotes the development of liver and/or kidney fibrosis through endocrine, cell signaling and other pathways. This paper reviews the research progress in understanding liver fibrosis and kidney fibrosis based on the gut-liver-kidney axis, which may be helpful for providing new strategies and theoretical basis for the diagnosis and treatment of hepatic and renal fibrosis.

Prostate cancer is one of the common malignant tumors of male urogenital system, and the incidence of prostate cancer in China has increased significantly in the past decade. At present, endocrine therapy based on androgen blockade is the main method of clinical treatment except radical surgery and radiotherapy/chemotherapy for prostate cancer. However, the clinical benefit can only be obtained in the early stage of treatment, and nearly90% of patients will develop to the castration resistance, and among them, nearly 90% of patients will have bone metastasis. The quality of life decreases sharply with the progression of disease for patients. In addition to the androgen signal pathway, studies have shown that many other oncogenic signal pathways have involved in the development of castration resistance, including classic cancer signaling pathways, immune and inflammatory signaling pathways, etc. Understanding the mechanism of androgen independent signal pathway in the formation of castration resistance will help to understand the off-target effect of androgen blocking therapy and introduce new treatment targets or strategies to get rid of the "no drug available" dilemma for clinical treatment of castration resistance.

Depression is a serious mental illness with a high incidence. At present, we do not fully understand the specific pathological mechanisms of depression, and the efficacy of drug treatments is very limited. Recent studies have shown that epigenetic changes that occur in specific brain regions may be a key mechanism by which environmental factors to interact with individuals to influence the risk of depression. Therefore, drugs that target epigenetic regulation may become a new direction for the development of antidepressants. Histone deacetylase inhibitors (HDACi) are a class of compounds that inhibit histone deacetylase activity, which has been reported to be associated with depression; this article addresses the use of HDACi in preclinical studies, and their potential therapeutic role and limitations of use in depression.

Chronic kidney disease(CKD) is a serious chronic disease with high incidence, poor prognosis, and a variety of complications. Indoxyl-sulfate(IS) and p-cresol sulfate(PCS) are two typical gut-derived uremic toxins, which are produced by the co-metabolism of intestinal microbes and the host. With the progression of CKD, gutderived uremic toxins such as IS and PCS accumulate in patients with CKD and thereafter accelerate the progression of CKD. Gut microbiota is closely related with CKD, and targeting gut microbiota to regulate gut-derived uremic toxins synthesis and metabolic pathways may be a promising strategy to delay the progression of CKD. In this paper, the relationship between gut microbiota, gut-derived uremic toxins, and CKD was analyzed, and the strategy to delay the progression of CKD by targeting gut microbiota and uremic toxins metabolism pathway was proposed.

The function of circulatory system, nervous system and endocrine system is significantly changed in hypoxic environments. These changes affect the absorption, distribution, metabolism, and excretion of drugs in the body. Drug metabolizing enzymes and transporters are the main factors affecting drug metabolism; microRNA(miRNA) can act directly on drug metabolizing enzymes and transporters and can regulate their genes through hypoxia-inducible factor, inflammatory cytokines, and nuclear receptors. This article reviews the effect of hypoxia on drug metabolizing enzymes and transporters and the mechanisms by which miRNA modulates these proteins and their expression during hypoxia.

Hyperlipidemia is a common disease with abnormal blood lipids and is an important risk factor for various cardiovascular diseases. Traditional Chinese medicine has the advantages of dependable lipid-lowering effects with few side effects and is widely used in the prevention and treatment of hyperlipidemia in China. However, due to the complex composition of traditional Chinese medicine and the many targets for treating hyperlipidemia, the mechanisms by which these medicines lower lipid levels are not well resolved. Lipidomics is a discipline that studies lipids and the interaction of lipids in biological systems. Lipidomics can identify and quantify the lipids in vivo under physiological and pathological conditions, helping to discover the potential biomarkers related to the lipid-lowering effects of traditional Chinese medicine and providing a basis for systematically studying the lipidlowering effect of traditional Chinese medicine. This review introduces the principal research methods used in lipidomics and summarizes the results and prospects of application of lipidomics in the research on the lipid-lowering effects of traditional Chinese medicine.

Chronic obstructive pulmonary disease and asthma are complex inflammatory diseases with airway obstruction as the main characteristics, and have become common respiratory diseases that seriously affect human health. Compared with the clinical use of PDE3 or PDE4 inhibitors alone, dual PDE3/4 inhibitors have synergistic anti-inflammatory and bronchodilator effects, and have attracted widespread attention in recent years. This article reviews the representative research results of dual PDE3/4 inhibitors currently in the preclinical and clinical research stages, summarizing their latest progress and their potential for the treatment of chronic obstructive pulmonary disease and asthma.

Cardiovascular disease is a principal cause of morbidity and death in the world. Although drug therapy has made great progress in the past few decades, there are still many deficiencies in the prevention and treatment of cardiovascular disease. Dyslipidemia is still a common risk feature and is not sufficiently controlled. A growing body of evidence suggests that the occurrence and development of cardiovascular disease is associated with many associated risk factors, such as higher low-density lipoprotein levels, lower high-density lipoprotein levels and high triglyceride levels. A number of clinical trials in patients with dyslipidemia have shown that actively decreasing low density lipoprotein cholesterol can significantly decrease cardiovascular events. ATP citrate lyase(ACLY) is a cytoplasmic homo-tetrameric enzyme. In the presence of adenosine triphosphate(ATP), ACLY catalyzes the conversion of citric acid and coenzyme A to acetyl-CoA and oxalyl acetate. ACLY is the main enzyme for the production of cytoplasmic acetyl-CoA, and cytoplasmic acetyl-CoA is the precursor required for de novo synthesis of cholesterol and fatty acids. Therefore, it is possible to reduce the production of acetyl-CoA and reduce the levels of cholesterol and triglycerides by inhibiting ACLY. ACLY can be used as a molecular target for reducing blood lipids, and there are an increasing number of studies on ACLY inhibitors. In this paper, the structure and mechanism of ACLY and its relationship with lipid metabolism are briefly introduced, and we review some current ACLY inhibitors.

The incidence of malignant melanoma, a highly fatal skin tumor, is on the rise worldwide. Melanomas are highly aggressive and have strong metastatic capability that leads to lethality. Recurrence occurs in patients with distant metastases, even with the latest treatments, and median survival is only a few months. At present, the prevention and treatment of melanoma includes surgical resection, chemotherapy, immunotherapy and targeted therapy. However, these strategies can lead to drug resistance and adverse effects. In recent years, an increasing number of studies have found that natural products have effective anti-melanoma activities, including inhibition of tumor growth, induction of cell apoptosis, inhibition of angiogenesis and metastasis and toxicity to tumor stem cells.In addition, several studies have reported that the combination of natural products and traditional anti-melanoma drugs can enhance the therapeutic efficacy. In this review we summarize the prevention and treatment of melanoma with natural products.

The determination and characterization of solid drug form polymorphisms plays an important role in drug quality control, selection of the production process and clinical efficacy evaluation. Vibrational spectroscopy is a powerful method for the characterization of drug polymorphisms. In this paper we review recent research and application advances in the polymorphic characterization of active pharmaceutical ingredients(APIs) and drug cocrystals/salts by using Fourier transform infrared(FTIR) and Raman spectroscopy to elucidate the characteristics of APIs and drug complexes. This may provide theoretical support for structural analysis during the development process for drugs.

Research on the material basis of Chinese materia medica(CMM) is the basis for modernization of CMM. High-resolution mass spectrometry(HRMS) has become a powerful tool for the qualitative analysis of the components of CMM. Some data-mining strategies based on high-resolution mass spectrometry, such as full-information tandem mass spectrometry scanning acquisition strategy, sequential windowed acquisition of all theoretical fragment ions, mass detect filter, characteristic ion filter, mass spectral tree similarity filter, etc. have greatly promoted the elucidation of the qualitative basis of CMM. In order to provide methods for the rapid discovery and structural characterization of components and metabolites of CMM, this review summarized the advances in HRMS-based data-mining technologies for detecting and characterizing the compounds and metabolites of CMM, which includes class compounds, all compounds and metabolites.

With the implementation of the two-child policy and the growing demand for child health, pediatric medication has been arousing widespread social concern. To develop the drugs suitable for children, including new compounds, new specifications and new dosage forms, is urgently required for pharmaceutical researchers. In this review, several technical bottlenecks for pediatric oral liquid preparations, as well as the novel strategies involved in drug nanocrystals, self-microemulsion, ion exchange resin and Pickering emulsion were discussed, which may be benefit to play a theoretical guiding role in the research and development of children's oral liquid formulation.

Nanomedicine has great potential in cancer therapy, but the complex tumor microenvironment greatly prevents nanomedicine from being effectively delivered into tumor in vivo. It has been widely accepted that the encapsulated drugs in the nanoparticles have to go through five major cascading steps, including blood circulation, accumulation in tumor, penetration into the depth of tumor tissue, internalization by tumor cells and then intracellular drug release, before they can exert the anti-tumor efficacy. Among the five steps, drug accumulation in tumor and penetration in the depth of tumor have been the two major issues undermines the antitumor efficacy of nanomedicine. This paper summarizes the new major progress in improving the tumor accumulation and penetration of nanomedicine, especially the technologies that appeared or developed in the last five years.

Natural deep eutectic solvent(NDES) is a kind of deep eutectic solvents(DESs) which is composed of natural substances with good biocompatibility. Those substances can function as hydrogen bond donor and acceptor, such as choline, amino acids, sugars, etc. NDES have been widely used in many fields due to their advantages of low cost, easy preparation and environmental friendliness. It is especially suitable for the pharmaceutical industry because of its good biocompatibility and safety for use. In this paper, we firstly review the molecular simulation methods for current design of DESs from the formation principle. And then, the materials and preparation of NDES are reviewed and the physicochemical properties are further described. Finally, we review the current application of NDES in pharmaceutics including increasing drug solubility, promoting drug permeability and enhancing oral drug absorption, and meanwhile their future applications in pharmaceutics were also prospected.

As a depot drug delivery system, injectable polylactide-polyglycolide(PLGA) sustained-release microspheres have been successfully used to treat many diseases since the first microsphere product Lupron depot was approved for marketing in the United States in 1989. It has the ability of long-term release in the body for several days to several months, which can not only reduce the times of administration, but also reduce the drug blood concentration fluctuations, significantly improve the safety and patient compliance. In vitro-in vivo correlation(IVIVC) makes the development of microspheres more possible. It can describe the dynamic information of drug release in vivo through the in vitro release behavior of microspheres, and can reduce the workload of each stage and shorten the time span while characterizing the performance of microspheres. IVIVC can provide guidance or support for drug development, production changes, supervision and management. This article summarizes the release mechanism of injectable PLGA sustained-release microspheres, common measurement methods and theories of in vitro and in vivo release. And we also focus on the establishment and application of IVIVC, especially A level IVIVC in the field of microsphere preparations, to provide a reference for further study on in vitro-in vivo correlation of microspheres.

Central composite design(CCD) is one of the most commonly used design methods in response surface optimization and has been widely applied in the field of pharmaceutics to optimize preparations. On the 20 th anniversary of the introduction of CCD into China, the paper reviews its application in domestic pharmaceutical researches. Based on the brief introduction of basic principle and operation steps of CCD, the mistakes emerging in the application of CCD are summarized, including conceptual confusion with Box-Behnken design and facecentered CCD as well as wrong designs. Besides, the issues concerning the selection of factors and responses are discussed. The article is helpful for researchers to comprehensively understand the CCD and facilitates the rational application of this method.

The emerging nano-black phosphorus materials have created a new platform for biomedical research. Nano-black phosphorus has the following advantages: black phosphorus can produce singlet oxygen under near-infrared light irradiation, so it can be used as a photosensitizer for photodynamic therapy; black phosphorus has extensive light absorption in the long wavelength region, and this near-infrared photothermal property can be used in photothermal therapy. The high specific surface area and unique fold structure of the black phosphorus nanosheet make it have very high drug loading.This paper mainly reviews the applications of black phosphorus in biological imaging, photothermal therapy, photodynamic therapy, and as a drug carrier in recent years. Based on the photoelectric properties of black phosphorus nanomaterials combined with intelligent drug delivery platform, the synergistic effects of light/heat/chemistry, light/chemistry/gene, and light/chemistry/immunity can be produced, which has a broad application prospect.

Fatigue is a common complication of type 2 diabetes mellitus (T2DM).We examined the relationship between T2DM fatigue and the skeletal muscle 5-hydroxytryptamine (5-HT) system.In animal experiments, a T2DM model was established in mice by feeding a high-fat diet with intraperitoneal injection of streptozotocin.The mice were treated with the 5-HT_2A receptor antagonist sarpogrelate hydrochloride (SH) and the 5-HT synthesis inhibitor carbidopa (CDP)(separately and in combination).In cell culture experiments, C2C12 cells were stimulated with D-glucose, palmitic acid or 5-HT.5-HT_2AR, 5-HT synthesis and 5-HT degradation were inhibited by SH, CDP, or monoamine oxidase A (MAO-A) inhibitor.The animal experiments were in accordance with the regulations of the Animal Ethics Committee of China Pharmaceutical University.The results showed that 5-HT_2AR, 5-HT synthase and MAO-A were expressed in mouse skeletal muscle and C2C12 cells.The expression of these proteins was significantly up-regulated in T2DM mice or when C2C12 cells were exposed to palmitic acid and D-glucose; palmitic acid was a stronger stimulant of their expression than D-glucose.Rotating rod experiments and biochemical index tests have shown that T2DM fatigue is associated with an increase in skeletal muscle 5-HT_2AR, 5-HT synthesis and 5-HT degradation.5HT_2AR mediates the expression of MAO-A and the synthesis of 5-HT, which indirectly regulates the degradation of 5-HT.MAO-A regulates cell inflammation, mitochondrial ROS production and membrane potential depolarization by mediating 5-HT degradation.MAO-A also inhibits the expression of peroxisome proliferator-activated receptor γ coactivator-1 (PGC-1), carnitine palmitoyltransferase-1 (CPT1) and ATP synthase-6 (ATP6), thus inhibiting mitochondrial functions such as fatty acid β oxidation and ATP synthesis.SH and CDP can effectively treat T2DM fatigue, and can also reduce blood glucose and blood lipid, and the combination of SH and CDP has a clear synergistic effect.

The study is to investigate the effect of glaucocalyxin A(GLA) on mast cell-mediated anaphylaxis.The animal welfare and experimental process of this experiment followed the regulations of the Animal Ethics Committee of Yanbian University. BALB/c mice were used in the animal experiment and randomly divided into five groups, control group, model group, and GLA low, medium, and high dose groups(10, 20, and 40 mg·kg^-1). Mice were sensitized by intradermal injection of anti-dinitrophenyl-immunoglobulin E(DNP-IgE) into the ears and challenged with a mixture of DNP-human serum albumin(HSA) and 4% evans blue into the tail veins to prepare an animal skin passive cutaneous anaphylaxis(PCA) model, which was collected from both ears for measurement of dye staining and histology. Rat peritoneal mast cells(RPMCs) were used in the cell experiment and divided into control, IgE + antigen(Ag), and IgE + Ag + GLA groups to determine histamine release as well as calcium influx levels. High-affinity IgE receptor(FcεRI)-mediated signaling pathway proteins and HMGB1/TLR4/NF-κB(high mobility group box 1/toll like receptor 4/nuclear transcription factor kappa B) signaling proteins were detected by Western blot. The results of animal experiments suggest that GLA inhibits PCA, reduces evans blue dye exudation, and reduces ear inflammation and ear thickness in mice. The results of cellular experiments suggested that GLA could reduce histamine release and calcium influx, and inhibit tumor necrosis factor-α(TNF-α), interleukin(IL)-4, IL-13, and IL-1β production; Western blot results showed that GLA inhibited FcεRI-mediated phosphorylation levels of spleen tyrosine kinase(Syk), Lck/Yes novel tyrosine kinase(Lyn), tyrosine kinase Fyn(Fyn), growthfactor receptor-bound protein 2(Gab2), and phospholipase C(PLC) γ1, while GLA inhibited HMGB1/TLR4 signaling pathway to limit NF-κB p65 nuclear metastasis. The results indicate that GLA inhibits mast cell degranulation and attenuates allergic inflammation through the HMGB1/TLR4/NF-κB signaling pathway.

In the treatment of hypertensive crisis, the novel Rho kinase inhibitor DL0805-2 can rapidly lower systematic blood pressure, reduce pulmonary artery pressure, and has a significant protective effect on lung injury.This experiment intends to evaluate the efficacy of DL0805-2 against pulmonary arterial hypertension (PAH) and preliminarily reveals its underlying mechanism. Animal welfare and experimental procedures are in accordance with the provision of the Animal Ethics Committee of the Institute of Materia Medica, Chinese Academy of Medical Sciences. Sprague Dawley (SD) rats were randomly divided into DL0805-2 low, medium, and high dose groups(1, 3, and 10 mg·kg^-1), bosentan positive control group, model group, and blank control group. The drug was administered daily on the 7th day after model establishment by monocrotaline injection. On the 25th day of the experiment, relevant indicators were examined to observe the therapeutic effect of DL0805-2 on pulmonary hypertension. DL0805-2 significantly relieved the abnormal changes in the physiological parameters related to PAH induced by monocrotaline, including reducing right ventricular systolic pressure, alleviating cardiac damage caused by pressure overload, and reducing the levels of endothelin-1 and inflammatory factors in lung tissues.DL0805-2 also attenuated pulmonary arteries remodeling. It was preliminarily discovered that DL0805-2 exerts preventive and therapeutic effect on PAH through Rho-kinase pathway. Our results suggested that DL0805-2 had good therapeutic effects on monocrotaline-induced PAH rat model. It intervened early in the disease process, effectively prevented the development of the disease, and reduced the mortality of the diseased animals. The mechanism is related to Rho-kinase pathway.

In this study, we investigated the inhibitory effect of SYT-1, a new compound of tetrahydroisoquinoline, on tumor cell proliferation and underlying mechanisms. Cell counting kit-8(CCK-8) method was used to detect cell proliferation; clone formation experiment was used to detect cell clone formation ability; JC-1 probe was used to detect cell mitochondrial membrane potential; 2', 7'-dichlorodihydrofluorescein diacetate(DCFH-DA)probe was used to detect intracellular reactive oxygen species; Annexin V-FITC/PI(fluorescein isothiocyanate/propidium) counterstaining method was used to detect apoptosis; Western blot assay was used to detect the expression level of related proteins. The experimental results show that SYT-1 has a significant inhibitory effect on the proliferation of six human-derived cancer cells. Among them, the inhibitory effect on breast cancer MCF-7 cells is the strongest, the half maximal inhibitory concentration(IC₅₀) of SYT-1 of 48 h administration on MCF-7 cells is 5.87 μmol·L^-1, which is better than that of cisplatin(8.92 μmol·L^-1). Further studies have shown that SYT-1 can dose-dependently inhibit the monoclonal formation ability of MCF-7 cells, and can cause the mitochondrial membrane potential of the cells to decrease and the level of reactive oxygen species to increase. In addition, SYT-1 can significantly inhibit the activation of PI3 K-Akt(phosphatidylinositol 3-kinase/protein kinase B) signaling pathway and induce apoptosis of MCF-7 cells. The above research results show that, as a new type of tetrahydroisoquinoline compound, SYT-1 has the potential to inhibit tumor cell proliferation.

This study aims to investigate the effect of baicalein on the metastasis of esophageal squamous cell carcinoma(ESCC) cells, and to elucidate the potential molecular mechanisms. Wound healing and Transwell migration and invasion assays were performed to detect the effect of baicalein on the migration and invasion of EC9706 and KYSE30 cells; the nude mice models of lung metastasis were applied to examine the function of baicalein in metastasis of ESCC by using KYSE30 cells. All animals were received humane care according to the Institutional Animal Care Guidelines approved by the Experimental Animal Ethical Committee of Henan University. Western blot assay was used to detect the protein levels of ERK/ELK-1/Snail signaling pathway. The data showed that baicalein significantly inhibited the migration and invasion of EC9706 and KYSE30 cells; Mechanistically, baicalein treatment led to a dramatically reduced expression of phosphorylated extracellular signal-regulated kinase 1/2(p-ERK1/2, T202/Y204), p-ETS-domain containing protein-1(p-ELK-1, S383), Snail, N-cadherin, and Vimentin, and a statistical increase of E-cadherin expression in EC9706 and KYSE30 cells; Furthermore, the inhibition of ERK1/2 by U0126 or siRNA remarkably enhanced the effect of baicalein on the above proteins. In summary, baicalein probably inhibits the migration, invasion, and metastasis of ESCC cells via blocking the ERK/ELK-1/Snail signaling pathway.

As a Ginkgo biloba extract preparation, shuxuening injection has a unique advantage in the prevention and treatment of acute and subacute stroke, but its main active ingredient is still unclear. Using a subacute model of stroke in mice constructed earlier, we further explored the contribution and mechanism of the two main components of total ginkgo flavonol glycosides and total ginkgolides in facilitating the neurofunctional recovery in stroke-induced mice. The pharmacodynamics was mainly evaluated by neurobehavioral changes, cerebral infarction volume, blood-brain barrier permeability and brain edema. The pathway and targets were predicted by transcriptome and network pharmacology. Finally, the mechanism was verified at the mRNA and protein levels. The results showed that the beneficial effect of total ginkgolides was greater than that of total ginkgo flavonol glycosides in both the pharmacodynamics and the regulatory mechanism of granulocyte adhesion and diapedesis involving granulocyte colony-stimulating factor(G-CSF), macrophage-1 antigen(MAC-1) and E-selectin. These findings suggest that shuxuening injection may improve the prognosis for mice with subacute stroke by down-regulating GCSF-mediated granulocyte adhesion and diapedesis pathway mainly through the total ginkgolide components. This finding is expected to provide reference for optimizing prescription and searching for natural drugs for targeting the treatment of ischemic stroke prognosis. The animal experiments in this study followed the regulations of Animal Ethics Committee of Tianjin University of Traditional Chinese Medicine.

Network pharmacological approaches were used to predict the components, targets and pathways of Erhuang decoction (EhD) in the treatment of acute lung injury (ALI). The SwissTargetPrediction platform, DisGeNET, Therapeutic Target Database (TTD), GeneCards and Online Mendelian Inheritance in Man (OMIM) databases were used to predict potential targets of EhD and were integrated with the predicted targets for the treatment of ALI. A protein-protein interaction network model was constructed by using String database and Cytoscape software; the DAVID platform was used for Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. A network of drug components-targets-pathways was constructed by Cytoscape software and the SwissDock platform was used to dock the molecules of EhD found in blood with the key disease targets. An ALI model was established in mice and inflammatory factor detection and Western blot protein expression experiments with lung tissue sections were carried out to verify the effect of EhD in the treatment of ALI. Animal experiment ethical requirements were approved by the Ethical Committee Experimental Animal Center of Shandong University (Grant Number: 2016020). We identified 148 potential targets including signal transducer and activator of transcription 3 (STAT3), vascular endothelial cell growth factor A (VEGFA), RAC-alpha serine/threonine-protein kinase (AKT1), and nuclear factor-kappa B/p65 (RELA). The potential targets are largely associated with the biological processes of inflammation, oxidative stress, and apoptosis. Additional pathways relate to cancer, VEGF signaling, mitogen-activated protein kinase (MAPK) signaling, and Toll-like receptors (TLRs) signaling, along with other signaling pathways. Pharmacodynamic experiments showed that EhD could significantly reduce the content of inflammatory factors and the degree of lung injury of ALI mice. Western blot revealed that EhD could significantly decrease the expression of NF-κB/p65 and upregulate the expression of NF-kappa-B inhibitor alpha(IκBα). From the perspective of network pharmacology, the mechanisms of EhD in the treatment of ALI is consistent with the characteristics of multiple ingredients, multiple targets and multiple pathways. This research provides a reference for further study of the mechanism of this traditional Chinese medicine.

We used metabolomics technology to identify and understand the biomarkers and therapeutic mechanisms of umbilical compress therapy based on Xiaozhang Tie(XT) to provide scientific evidence for its clinical application. A total of 10 patients with cirrhotic ascites and gastrointestinal motility disorders who were hospitalized in the Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine from November 2017 to June 2018 were divided into a placebo group(4 cases) or an XT group(5 cases), and 10 healthy volunteers were included as controls. This clinical trial was approved according to the Ethics Committee of Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine(2017-528-11-01). The patients in the XT group were given umbilical compress therapy with Xiaozhang Tie, and patients in the placebo group were administered a plaster patch in which the drug content was less than 5%, receiving one patch per day for three consecutive days.Non-targeted metabolomics technology and UPLC-Q/Orbitrap-MS/MS analysis technology were utilized to investigate the fluctuations in endogenous metabolic profiles in the patient's urine prior to and after administration of XT. By analyzing and comparing the urine metabolic profiles of patients with cirrhotic ascites to those of healthy volunteers, a total of 31 biomarkers were identified, 14 of which were significantly decreased by the intervention with Xiaozhang Tie(P < 0.05). Pathway enrichment analysis revealed that phenylalanine metabolism and tryptophan metabolism are key pathways affected by XT treatment. The results suggest that XT can alleviate cirrhotic ascites by modulating abnormalities in amino acid metabolism.

Xianling Gubao is a common and effective medicine in the treatment of orthopedic diseases. In recent years, it has been reported to be associated with liver injury. However, through the analysis of the adverse drug reaction reports and key hospital cases, we found that there is considerable incomplete information in the reports of Xianling Gubao-related liver injury cases retrieved from the literature. Thus, it is difficult to accurately judge causality between the drug and liver injury. Six cases of liver injury related to Xianling Gubao were identified in key hospitals, two of which achieved the clinical diagnosis according to the assessment of the integrated evidence chain method. We further analyzed the public health data of all residents in Yinzhou. The gross incidence rate of Xianling Gubao-related liver injury was 0.034%, which corresponds to a level of rare incidence. This revealed that Xianling Gubao-related liver injury has significant divergence in individuals and an idiosyncratic nature. The gross incidence of liver injury related to Xianling Gubao was lower than that of other medicines for the treatment of orthopedic diseases. Based on the idiosyncratic drug-induced liver injury model mediated by immune stress, it was found that Epimedii Folium and Psoraleae Fructus were the major components that lead to liver injury, and the liver injury caused by a full prescription was less serious than that encountered with only Epimedii Folium and Psoraleae Fructus. This suggests that the other 4 herbs(Dipsaci Radix, Anemarrhenae Rhizoma, Rehmanniae Radix, Salviae Miltiorrhizae Radix et Rhizoma) can prevent/alleviate the liver injury. Through disassembled prescription analysis, we found that the attenuation efficacy of Salviae Miltiorrhizae Radix et Rhizoma was the most significant.In conclusion, Xianling Gubao may cause idiosyncratic liver injury in a tiny minority of susceptible individuals, but the incidence risk is lower than that of other commonly used drugs for orthopedic disease. Xianling Gubao should be discreetly applied to patients with immune stress. The major components that induced liver injury in Xianling Gubao were Epimedii Folium and Psoraleae Fructus, and Salviae Miltiorrhizae Radix et Rhizoma appears to attenuate this toxicity. This study provides a reference for the rational clinical medication with Xianling Gubao.

Seventy-nine injectable herb extractions have been approved by the Chinese Food and Drug Administration(CFDA) and are frequently administered intravenously for various diseases. Unfortunately, herb-drug interactions are under-investigated and sometimes overlooked in the clinic. In the present investigation the in vitro inhibition of 9 drug metabolizing enzymes including CYP1A, CYP2A6, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP2E1, and CYP3A was assessed using an appropriate probe substrate for each enzyme with human liver microsomes. Metabolite formation was quantified using a validated and sensitive high-performance liquid chromatography-tandem mass spectrometry(LC-MS/MS) assay. The IC₅₀ of each herb extract was estimated using a concentration range from 5% to 0.5%, and the time-dependent inhibition of the nine CYP450 isoenzymes was also determined. Of the 79 approved iv herb injectables, 37 inhibited CYP1A, 24 inhibited CYP2A6, 41 inhibited CYP2B6, 36 inhibited CYP2C8, 31 inhibited CYP2C9, 41 inhibited CYP2C19, 13 inhibited CYP2D6, 25 inhibited CYP2E1, and 42 inhibited CYP3A with 50% or greater inhibition at a test concentration of 5%(v/v). IC₅₀ differences were noted between pre-incubation or co-incubation assays with HLM for 30 min, with the time-dependent inhibitory(TDI) effects were observed with 2 injectables on CYP1A, 5 injectables on CYP2A6, 5 injectables on CYP2B6, 6 injectables on CYP2C8, 1 injectable on CYP2D6 and 6 injectables on CYP3A. Collectively, the results demonstrate that potential herb-drug interactions(HDIs) can occur with the concomitant use of herb injectables and prescription drugs that are cleared by CYP450 enzymes, and further investigation is warrant for the clinical relevance of these interactions.

Eight polyacetylenes were isolated from the extract of the stems and leaves of Chrysanthemum morifolium by various chromatographic methods. Their structures were determined as 2E, 4E, 12Z-tetradecatriene-1-pyrrolidine-1-oxo-8,10-diynoic(1), tetradeca-2E, 4E, 12E-trien-8,10-diynoic acid pyrrolidide(2), tetradeca-2E, 4E-dien-8,10-diynoic acid pyrrolidide(3), tetradeca-2E, 4E, 10Z-trien-8-ynoic acid pyrrolidide(4), 2E, 4E, 12E-tetradecatriene-8,10-diynoic acid isobutylamide(5), 2E, 4E-undecyldiene-8,10-diynoic acid isobutylamide(6), 2E, 4E, 10E-N-isobutyl-2,4,10-tetradecatrien-8-ynoic acid amide(7), and undeca-2E, 4E-diene-8,10-diynoic acid phenylethylamide(8) by spectroscopic methods, including UV, IR, ESI-MS, HR-ESI-MS, 1D and 2D NMR spectra.Among them, compound 1 is a new polyacetylene, and compounds 2-8 were isolated from this plant for the first time. Compounds 5-8 inhibited the proliferation of A549 cell significantly at certain concentration, showing potent antitumor activity.

Ten triterpenoid saponins were isolated and purified from the water extract of Glycyrrhiza glabra by polyamide resin combined with macroporous resin column chromatography, ODS medium pressure column chromatography and semi-preparative RP-HPLC. Their structures were elucidated by physicochemical properties, NMR and MS spectra, and determined as 3β-O-[β-D-glucuronpyranosyl-(1→2)-β-D-glucuronpyranosyl]-30β-O-β-D-glucuronpyranosyl-oleanane-11-oxo-12(13)-ene(1), 3β-O-[β-D-glucuronpyranosyl-(1→2)-β-D-glucuronpyranosyl]-30 β-O-α-L-rhamnopyranosyl-oleanane-11-oxo-12(13)-en-22 β, 30-diol(2), uralsaponin C(3), licorice-saponin A3(4), licorice-saponin P2(5), 22β-acetoxyl-glycyrrhizin(6), macedonoside A(7), 29-hydroxyl-glycyrrhizin(8), licorice-saponin G2(9), glycyrrhizin(10). Compounds 1 and 2 are two new compounds and named as licorice-saponin R3 and licorice-saponin S3.

The quality markers(Q-markers) of traditional Chinese medicine(TCM) have become a topic of interest in TCM research in recent years. Nonetheless, there is still no consensus on how to scientifically characterize TCM Q-markers. Our study establishes an identification method for TCM Q-markers based on the analytical hierarchy process(AHP) and the entropy weight comprehensive method. By constructing an evaluation system encompassing the target layer, the factor layer and the control layer, AHP can be used to analyze the weight of three core TCM quality attributes, including effectiveness, testability and specificity. Following that, the entropy weight method is employed to analyze the specific indicators for each attribute based on the literature and experimental data. Finally, the comprehensive weight of each index is obtained by combining the two weights, and the comprehensive weight and the specific value of each component is multiplied and summed to obtain the integrated score ranking, and thereby identify the TCM Q-markers. Taking Shaoyao Gancao decoction as an example, the analysis revealed that the top 8 components are as follows: paeoniflorin > quercetin > albiflorin > glycyrrhizic acid > naringenin > liquiritin > oxypaeoniflorin > benzoylpaeoniflorin, and can be identified as Q-markers of Shaoyao Gancao decoction. This study not only provides support for the establishment of quality standards and process quality control of TCM formulae, but also provides innovative ideas and methods for quantitative evaluation and accurate identification of TCM Q-markers.

To improve the efficacy of 5-aminolevulinic acid(5-ALA)-mediated photodynamic therapy(PDT), a fluorocarbon microemulsion-based gel(FMBG) loaded with both 5-ALA and carbon dioxide(CO₂) was prepared in this study. Its physical and chemical properties such as particle size, zeta potential, morphology, pH value and viscosity were characterized. Acid-base titration experiment was used to determine the CO₂ loading, a fluorescence derivatization method was established to determine the content of 5-ALA, and the confocal laser scanning microscope and Franz diffusion cell method were carried out to investigate its transdermal ability. Through the laser speckle contrast imaging, the CO₂-affected blood flow perfusion of skin was measured. Finally, the skin irritation test was tested by hematoxylin-eosin staining(H & E) method. These results showed that the prepared FMBG was a milky white gel, with an average particle size of 202.4 nm, a zeta potential of-25.3 mV, a pH of 6.0, and a viscosity of 1 062.0 mPa·s. It can be stored stably for seven days at room temperature. The 5-ALA content of FMBG was measured to be approximately equal to 20%(w/w). At room temperature and normal pressure, the CO₂ loading content of FMBG was 5.016 mg·L^-1, which was 1.5 times as much as that of water. The transdermal absorption experiment and blood perfusion results showed that the FMBG can effectively enable the transdermal delivery of 5-ALA and CO₂, and significantly increased the blood perfusion of skin. H & E staining results indicated that FMBG had negligible skin irritation(all animal tests were approved by the Ethics Committee of 900 Hospital of the Joint Logistics Team). In this study, a safe and stable FMBG loaded with both 5-ALA and CO₂ was successfully prepared. It was suitable for transdermal application, having the potential of enhancing the efficacy of 5-ALAmediated PDT.

Under the guidance of Chinese Pharmacopoeia(2020 edition), the functionality-related characteristics of hydroxypropyl methylcellulose(HPMC) type 2208 from imported A manufacturer, domestic S manufacturer, domestic T manufacturer and different batches of the same manufacturer were characterized. The principal component analysis was used to comprehensively evaluate the functionality-related characteristics. The results were as follows: hydroxypropyl methylcellulose had no significant difference in viscosity and molecular weight distribution between different manufacturers, and there were significant differences in the cumulative particle size distribution of the sample reaches 50%(d₅₀) and 90%(d₉₀), bulk density, tap density and Carr's index. The HPMC from A manufacturer have the biggest inter-batch difference of particle size and their inter-batch difference of polydispersion coefficientis smaller than S manufacturer. Domestic manufactures have the largest inter-batch difference in other functionalityrelated characteristics. The three principal components were extracted by principal component analysis, and the variance contribution rate reached 89.44%, indicating that the extracted principal components can explain all the data well. By constructing a comprehensive evaluation model, the comprehensive score ranking of all HPMC samples is obtained: S manufacturer > A manufacturer > T manufacturer.

Chalcone isomerase (CHI) is the second rate-limiting enzyme involved in the biosynthetic pathway of flavonoids in Glycyrrhiza uralensis. Based on our previous studies, we selected the specific CHI haplotype (GenBank Accession No. KY115232) to maximize flavonoid accumulation. We constructed a plant binary expression vector for overexpression of this CHI gene by the gene fusion method and transfected the plasmid into Agrobacterium tumefaciens ACCC10060 by electroporation. The recombinant A. tumefaciens ACCC10060 subsequently was used to infect cotyledons and hypocotyls of G. uralensis to obtain transgenic hairy roots. A qRT-PCR method was used to determine the copy number of CHI and a UPLC method was used to assay the content of four flavonoids in different hairy root lines. The qRT-PCR results showed that the copy number of CHI in hairy roots was 1 or 5. UPLC results showed that the content of total flavonoids, liquiritin, liquiritigenin, and isoliquiritigenin in transgenic hairy root samples was significantly higher than that in wild-type samples. This study demonstrates that overexpression of CHI significantly increases the content of flavonoids in hairy roots of G. uralensis. This work provides a theoretical basis for clarifying the function of CHI. Three transgenic hairy root lines of G. uralensis were isolated which can be used to increase the accumulation of licorice flavonoids in vitro.

Rhamnose synthase (RHM) is a key enzyme in the biosynthesis of uridine diphosphate rhamnose (UDP-Rha), reversibly converting uridine diphosphate-glucose (UDP-Glc) into UDP-Rha in the presence of NADH or NADPH. In this research, yeast extract (YE) was used to stimulate Sorbus aucuparia suspension cells. Based on a previous study of the transcriptome database of S. aucuparia suspension cells, two RHMs were cloned from S. aucuparia and named SaRHM1 (GenBank No.: MK213340) and SaRHM2 (GenBank No.: MK213341). The SaRHM1 gene contained a 2 007 bpopen reading frame (ORF) encoding a polypeptide of 668 amino acids with a molecular weight of 75.25 kD, and a theoretical isoelectric point (pI) of 7.24. The SaRHM2 gene contained a 2 040 bpORF encoding a polypeptide of 679 amino acids with a molecular weight of 76.26 kD and pI of 6.41. Bioinformatic analysis indicated that SaRHM1 and SaRHM2 contained two special sequences of GxxGxxG/A and YxxxK. Multiple sequence alignments and phylogenetic trees show that SaRHM1 and SaRHM2 have high sequence similarity with other plant species of RHMs. The results of enzyme activity assays in vitro revealed that both recombinant SaRHM1 and SaRHM2 are able to convert UDP-Glc into UDP-Rha. SaRHMs displayed maximum activity at 40 ℃ and a pH of 8 and 9, respectively. The K_m values of SaRHM1 and SaRHM2 for UDP-Glc were 212.4 ± 56.70 and 361.0 ± 63.74 μmol·L^-1, respectively, with V_max values of 235.5 ± 18.98 and 516.5 ± 22.30 nmol·min^-1·μg^-1, respectively. This study reports the cloning and sequencing of RHMs from S. aucuparia and verifies their function, which likely provide rhamnose donors for the subsequent biosynthesis of rhamnosides.