In the era of genome-wide association study (GWAS), a large number of drug response-related loci have been identified in the non-coding sequences. The interpretation of these loci in mechanism is concerned with the effects on the mRNA expression level of these genes. Expression quantitative trait loci (eQTL) studies indicate the relationship of genome variants and the level of mRNA. Its elucidation of the relationship between genetic variation and gene expression, gene interaction and gene regulatory network provides an efficacious mean for pharmacogenomics. The effects of gene polymorphism on drug responses have been unraveled thoroughly in studies which combined pharmacogenomics with eQTL and GWAS.

Bromodomain and extraterminal (BET) proteins are a class of proteins that can interpret epigenetic codes and play an important role in regulating gene transcription through identifying and binding acetylated histones or non-histones proteins. The BET inhibitors have emerged with good therapeutic effects in preclinical disease models such as cancer and inflammation. Some of them have entered clinical studies, demonstrating that there is considerable prospect for drug development with BET as a potential therapeutic target. This review briefly describes the structures and functions of the BET proteins, the BET inhibitors in various diseases, as well as molecular mechanisms involved.

Fibronectin extra-domain B (ED-B) has been a good target in new drug development, several relevant antibody drugs are in phase Ⅱ or Ⅲ clinical trials for metastatic melanoma, soft-tissue sarcoma and so on. Some data of phase Ⅱ clinical trials shows that ED-B antibody drugs (L19-IL2 and L19-TNF α) for melanoma are significantly superior to PD-1 antibody drugs. This article describes several aspects of ED-B, such as biological characteristics, the development of targeted drugs, and the potential therapeutic applications, including modifying protein drug structure, constructing fusion protein, expanding indications, developing companion diagnostics and individual treatments. We also discuss how to promote original innovation in drug discovery, which might help to find new development focus.

To a certain extent, the drug effect is determined by its blood concentration. It is generally accepted that the blood concentrations of constituents of Chinese medicines are very low. There is no sufficient experimental bases and references on its degree and the possibility of taking effect. In this study, 69 papers were collected and analyzed by searching the database of Scifinder, Pubmed, CNKI. The minimum effective blood concentrations of 73 common Western medicines and the maximum blood concentrations of 211 in vivo constituents of 40 Chinese medicines (single herb or compound Chinese medicine) were summarized. It was found that the maximum blood concentrations of the most in vivo constituents of Chinese medicines were much less than the minimum effective blood concentrations of the Western medicines. Specifically, the minimum effective blood concentrations of 17 Western medicines (23% of total) and the maximum blood concentrations of the 143 in vivo constituents of Chinese medicines (68% of total) were less than 100 ng ·mL^-1; the minimum effective blood concentrations of 31 Western medicines (42% of total) and the maximum blood concentrations of the 20 in vivo constituents of Chinese medicines (9% of total) were more than 1 000 ng·mL^-1. In this paper, a systematic summary and comparison of the blood concentrations in traditional Chinese medicines and Western medicines were conducted, which could provide a new ideas and references for the study of the pharmacodynamical material basis and its mechanism in traditional Chinese medicine.

Poly(2-ethyl-2-oxazoline) (PEOz), a hydrophilic long-chain polymer synthesized by living cationic ring-opening isomerization polymerization of 2-ethyl-2-oxazoline, has the characteristics of low toxicity, biocompatibility, flexible chain, and modified expediently. PEOz is a potential substitute of polyethylene glycol (PEG) to render the ability of long-circulation, enhance cellular uptake and endosomal escape behaviors to PEOz modified drug delivery system. In this review, we summarized recent literature for the research progress of physicochemical properties, synthetic methods and the application of PEOz in drug delivery system.

Increasing evidence suggests that hepatocellular carcinomas (HCCs) are sustained by a distinct subpopulation of self-renewing cells known as cancer stem cells (CSC). However, our understanding of their regulation is limited. Rapid reversible changes of CSC-like cells within tumors may result from the effect of biological mediators found in the tumor microenvironment. This paper aims to explore how nitrite, a key cellular modulator whose level is elevated in many tumors, affects CSC-like phenotypes of human hepatoma cells SMMC-7721 cells. The SMMC-7721 cell line was cultured under serum-free conditions to produce floating spheres. The distribution of cell cycle was analyzed by flow cytometry, the capability of cells self-renew was detected by colony-forming capabilities and spheroid-formation assay, the expression of stemness protein such as CD133, CD90 and EpCAM were determined by flow cytometry and Western blot, cell invasion was analyzed by transwell assay, and viability of SMMC-7721 parental cells and spheroids cancer cells was determined by the 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide (MTT) assay. Xenograft tumor models were established by subcutaneously injecting SMMC-7721 spheroids cancer cells, the transplanted tumor tissue ROS levels was detected by reactive oxygen species (ROS) test kits, the expression of HIF-1α was observed by immunofluorescence. Our results showed that the SMMC-7721 spheroid cells were enriched with CSCs properties, indicated by the ability to self-renew, increased expression of CSCs markers, and increased resistance to chemotherapeutic drugs. Additionally, SMMC-7721 parental cells and spheroids cancer cells were treated with 150 μmol·L^-1 sodium nitrite for 6 days, compared with control cells, an increased accumulation of G₀/G₁ phase cells was observable in treatment cells. Indeed, our data demonstrated that in parent cells and spheres cells that were treated with sodium nitrite for different time, the cells' ability to chemoresistance and invasion, clone-forming efficiencies and the spheres forming ability were significantly higher than that of control cells. Exposure of sodium nitrite regulated CSC-like phenotype, indicated by increased expression of known CSC markers, CD133, CD90 and EpCAM in the exposed parental cells, as well as in dormant spheroids cancer cells. Compared with the parent cells, the above effects of nitrite on the spheres cells were significantly enhanced. In vivo data also presented a more significant promotion of tumor xenograft growth from the nitrite treatment than from either of the control. Mechanistic analysis indicated that nitrite induced the upregulation of HIF-1α as well as the downregulation of ROS in the tumor microenvironment. These results suggest that nitrite increases the invasiveness of SMMC-7721 cells through up-regulation of tumor stemness.

The impact of statins on airway inflammation has not yet been established and it may differ from their cholesterol-lowering effects. Oral administration of statins at large-doses may have adverse effects. It is possible to overcome the side effect to increase the clinical efficacy through the inhalation route. Female BALB/c mice were randomly divided into four groups including the control group (NS-vehicle), model group[ovalbumim (OVA)-vehicle], simvastatin (Sim) group and dexamethasone (DXM) group at 10 mice in each group. In this study, we hypothesize Sim as a potential anti-inflammatory drug with biological and pharmacokinetic properties suitable for delivery through the inhalation route. Mice were immunized with OVA and then challenged with OVA aerosol to induce the asthma reaction. Sim was inhaled at a dosage (5 mg ·mL^-1, ih, 15 min) or administrated by intraperitoneal injection (40 mg·kg^-1, ip) or gavage (40 mg·kg^-1, ig) during the OVA-challenge. In the mouse model of asthma, Sim significantly attenuated the total inflammatory cell counts and eosinophil counts (P < 0.01 or P < 0.05) via the different routes. Pretreatment with Sim at 1, 5, 20 mg·mL^-1, ih, significantly decreased the total inflammatory cell counts and eosinophil counts in alveolar lavage fluid (BALF) (P < 0.01) and the inhibitory effect was increased with the dosages of Sim via inhalation. Both of DXM and Sim at 5, 20 mg·mL^-1, ih, were more potent than that of Sim at 1 mg·mL^-1, ih. Sim significantly decreased IL-4 and IL-5 mRNA expression of lung at 5, 20 mg·mL^-1, ih (P < 0.01 or P < 0.05). Sim (5, 20 mg·mL^-1, ih) significantly decreased levels of IL-4 and IL-5 in BALF (P < 0.01 or P < 0.05). However, Sim (1 mg·mL^-1) declined slightly on IL-4 level in BALF. Sim at 5, 20 mg·mL^-1 had a greater rate of decline in IL-5 than at 1 mg·mL^-1. These results suggest Sim with different doses as a potential anti-inflammatory drug for airway inflammatory diseases with properties suitable for delivery by inhalation, which probably overcome the side effects and low clinical efficacy of oral Sim.

This study was designed to investigate the effect of minocycline on microglia activation of M1/M2 phenotypes. The model was induced by lipopolysaccharide (LPS) in BV-2 microglia cells, and was used to evaluate the effect and mechanism of minocycline. We measured nitric oxide (NO), prostaglandin E₂ (PGE₂), tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6) and interleukin-1 beta (IL-1β) in M1 type microglia, and interleukin-10 (IL-10) and transforming growth factor beta (TGF-β) in M2 type microglia through enzyme linked immunosorbent assay (ELISA). We used flow cytometry to detect the expression of M1 marker CD16/32 and M2 marker CD206 in order to evaluate the influence of minocycline on microglia activation of M1/M2 polarization. Finally, we explored the mechanism of minocycline through detection of the protein expression in response to activation of toll like receptor 4 (TLR4)-mediated myeloid differentiation factor 88 (MyD88) dependent pathway, mitogen activated protein kinase (MAPK) signaling and nuclear factor-κB (NF-κB). The results suggest that minocycline obviously inhibited the production of NO, PGE₂, TNF-α and IL-6, and increased the production of IL-10, TGF-β in LPS-stimulated BV-2 cells. Minocycline significantly down-regulated the expression of M1 marker CD16/32 and up-regulated the expression of M2 marker CD206. These results suggest that minocycline can inhibit the activation of microglia to M1 phenotype and promote the transformation of M2 phenotype through down-regulation of p38 and NF-κB signaling pathways.

This study was designed to investigate the synergistic analgesic effect between choline (Cho) and acetaminophen (Ace). Mice were treated with 0.6% acetic acid solution by intraperitoneal injection to build acetate writhing model. The KM mice were randomly divided into four groups:control group (n=10), Cho group (n=50), Ace group (n=50), combination group (Cho+Ace group, n=40), then the writhing times were counted respectively. OriginPro8.5 was used to calculate ED 50. The isobolographic analysis was used to test the interaction of Cho and Ace. To explore the mechanism, forty KM mice were randomly divided into control group, Cho group, Ace group and Cho + Ace group. Blood was collected for detection of TNF-α, IL-6, PGE₂ and NF-κB content using ELISA kits. The result ED 50 was calculated as followings. ED⁵⁰ of Cho and Ace was 19.47 mg·kg^-1 and 20.56 mg·kg^-1. The concentrations were 2.94 mg·kg^-1 for Cho and 3.15 mg·kg^-1 for Ace in the combination test. The levels of TNF-α, IL-6, PGE₂ and NF-κB in Cho group and Ace group were lower than those in the control group (P < 0.05). Compared to the Cho group and Ace group, the levels of TNF-α, IL-6, PGE₂, NF-κB in Cho + Ace group were reduced further (P < 0.05). The results revealed that Cho and Ace have synergistic analgesic effects, which may associate with inhibition of the NF-κB signaling pathway.

This study was designed to explore the "multi-components, multi-targets and multi-pathways" intervention mechanism of Huanglian Jiedu decoction (HLJDD) in the treatment of Alzheimer's disease (AD) by pharmacological network technology, which may establish a foundation for drug development and innovative research. Seventeen active constituents of HLJDD with anti-AD activities were submitted to PharmMapper and Molecule Annotation System (MAS 3.0) bioinformatics softwares to predict the target proteins and carry out related KEGG pathways annotation respectively. The network of "active compound-target-pathway" was constructed and analyzed using the Cytoscape 3.4.0 software. The results suggest that 47 pathways are affected by the 17 active components through 59 target proteins, in which 4 target proteins are related to AD and 2 pathways related to neuroinflammation, respectively. The effect of HLJDD on AD may be dependent on clearing/reducing β-amyloid protein, inhibiting Tau hyperphosphorylation, anti-inflammation and immunoregulation.

Hepatic disease is one of the high-prevalence diseases in China, of which gastrointestinal bleeding is a common complication treated by proton pump inhibitors. Vonoprazan is a novel proton pump inhibitor which acts better than lansoprazole in pharmacokinetics and pharmacodynamics. In this study, the pharmacokinetics of vonoprazan was compared between acute hepatic injury and normal condition in rats. Results showed that the exposure (AUC) of vonoprazan was significantly higher in rats with acute hepatic injury than in normal rats, and the metabolites formation rates of vonoprazan also slowed down, which might be due to the change of activity of enzymes and transporters. This find may provide a theoretical basis for the dose regulation of vonoprazan in patients with hepatic injury.

The glucose consumption activity of 9-substitued analogues of berberine was evaluated in L6 myotubes. It was found that the introduction of an ethoxy group on the 9-position of berberine was beneficial for the activity. 9-Ethoxy berberine analogue 2a exhibited superior activity to berberine in multiple dose levels, and the activity of 2a was 5.4 times as high as that of berberine at the dose of 1.25 μmol·L^-1. At the meantime, the potency on AMPK activation of 2a was 2.8 times of that of berberine at the dose of 10 μmol·L^-1. Therefore, the compound 2a is a promising scaffold for further modification.

In order to find highly active antidiabetic agents, the 3-amino group of skeletal structure of thiazolidine-2, 4-diones (TZDs) was modified to generate the new molecules TM1 and TM2 in the present research. The new molecules TM3-TM6 containing rhodanine structural units were designed based upon the bioisostere and combination principles. The target molecules TM7, which is similar to the traditional TZDs structurally, were designed by connecting the phenolic hydroxyl of the above target molecules to carbazole through a linker. All of these target compounds were synthesized successfully by selecting suitable synthetic routes with optimized procedures. The assay results of peroxisome proliferator activated receptor response element (PPRE) agonist activity revealed that the PPAR agonist activity was decreased due to the change of TZD ring. The assay of α-glucosidase inhibitory activity and protein tyrosine phosphatase-1B (PTP-1B) inhibitory activity showed that most of the seven serials target molecules have weak activities in vitro. However, 3 of the compounds exhibited strong PTP-1B inhibitory activities. TM2-6 exhibited the highest inhibitory activities, which reached 96.71% with IC₅₀ 1.48 mg·L^-1. In addition, the toxicity prediction disclosed that the highly active compounds were almost non-toxic. These results provide a hint for the development of new antidiabetic

The chemical constituents of Dioscorea opposite Thunb. were isolated and purified by Diaion HP-20, Sephadex LH-20, Toyopearl HW-40, MCI Gel CHP-20, ODS, silica gel column chromatography and semi-preparative-HPLC. Nine compounds were found and their structures were elucidated by spectral data and physicochemical properties, which were identified as 2-(1', 2', 3'-trihydroxybutyl-4'-O-α-D-glucopyranoside)-6-(2", 3", 4"-trihydroxybutyl)-pyrazine (1), uracil (2), xanthine (3), hypoxanthine (4), thymine (5), adenosine (6), uridine (7), inosine (8), and 5'-deoxy-5'-methylsulphinyladenosine (9). The compound 1 is a new pyrazine derivative, and the compound 3-5, 8, 9 are found in this plant for the first time.

Three sesquiterpenoids were isolated from the dichloromethane extract of the roots of Stelleropsis tianschanica Pobed through a combination of various chromatographic approaches, including silica gel, Sephadex LH-20, reverse phase C₁₈ and so on. On the basis of spectroscopic data analysis, they were identified as (+)-guaia-l(10), ll-dien-9-one-5α-hydroxy (1), 4β, 5βH-guai-9, 7(11)-dien-12, 8-olide-1α, 8α-diol (2), 4α, 5βHguai-9, 7(11)-dien-12, 8-olide-1α, 8α-diol (3). The compound 1 is a new sesquiterpenoid, and the compound 3 was isolated for the first time from the genus Stelleropsis.

The study is aimed to test the effect of Huangqin Tang (HQT) on serum metabolic profile in rats with ulcerative colitis, and explore its possible action mechanism for ulcerative colitis (UC) rats. The model of UC rats with cell immunoreactivity was made using a compound method (trinitrobenzene sulfonic acid plus ethanol). Rats were randomly divided into the control group, the model group, and HQT group. Ultra performance liquid chromatography tandem mass spectrometry (UHPLC-MS), principal component analysis (PCA) and partial least squares-discriminant analysis (PLS-DA) were employed to analyze the metabolic profile among normal group, the model group, HQT group. Potential biomarkers were screened in the serum based on the variable importance projection (VIP) value > 1, P < 0.05. As compared with the normal group, 16 potential biomarkers such as valine, tryptophan, lactic acid and urea were found and identified in the serum of model group rats. As compared with the model group, a part of the biomarkers were restored nearly to a normal state after HQT administration for 10 days. Metabolomic analysis revealed that the HQT has a certain therapeutic effect in UC rats, and the mechanism may be related to regulation of lipid metabolism, amino acid metabolism and energy metabolism.

To study the substances in fudosteine, one synthetic by-product and five forced degradation products were detected by hydrophilic interaction chromatography (HILIC). Quadrupole-time-of-flight mass spectrometry (Q-TOF MS) was used for accurate mass determination and product ion scanning. Five related substances were identified in the products of mass spectra fragmentations elucidation, and verified further according to synthetic process and stress testing results. The results obtained are valuable for fudosteine manufacturing process control and quality assurance.

With the extracts of Puerariae Lobatae Radix as the research object, the moisture absorption isotherm at 298, 308 and 318 K was determined through dynamic water vapor adsorption. Moisture absorption isotherm models were applied to the simulation of moisture absorption behavior. The enthalpy, entropy and Gibbs free energy for moisture adsorption were calculated according to the model and the enthalpy-entropy compensation was used to analysis moisture adsorption process. It was shown that the adsorption isotherm coincided with GAB model or Ferro-Fontan model. Moisture absorption process was an exothermic process which was driven by entropy. The effect of moisture on compressibility was evaluated through tensile strength and elastic recovery with water content, pressure force and speed variation. It was supposed that right amount of moisture was required to compression and compaction.

Fluorescent polystyrene nanospheres (PS) were used to explore the impact of substrate stiffness on cell uptake of nanoparticles in the breast cancer cells. Polyacrylamide (PAA) gels with varying stiffness were prepared by photopolymerization, and type Ⅰ rat tail collagen was covalently conjugated on the surface of PAA gels to facilitate cell adhesion. Type Ⅰ rat tail collagen was also used to fabricate collagen gels for 3D cell culture. Cells of human breast cancer cell line MCF-7 were incubated in the 2D culture on PAA gels and 3D culture within collagen gels. Next, nanospheres of 20 nm and 50 nm polystyrene were applied to MCF-7 cells in the 2D or 3D cultures. Cell morphology and uptake efficiency were observed with confocal microscopy. Our study demonstrates that substrate stiffness differentially regulated the cell morphology as well as the cell uptake behavior of polystyrene nanospheres in MCF-7 cells under 2D or 3D culture conditions.

The difference in pH between apical and basolateral side of intestinal epithelial and pH dependence character of the combination of FcRn (neonatal Fc receptor) and ligand might improve the delivery of hydrophobic drugs by facilitating the transcytosis of nanocarriers. Here we designed FcBP (IgG Fc domain-binding peptides) decorated coumarin 6 (C6) loaded poly(ethyl ethylene phosphate)-co-poly(ε-caprolactone) (PEG-PCL) micelles with different ligand densities to study the effect of pH and ligand density on the endocytosis and exocytosis process of micelles on human colon adenanocaricinoma cell lines (Caco-2). Active micelles with different ligand densities and passive micelles were prepared using the thin-film hydration method. The size of the micelles was characterized by dynamic light scattering analysis and the morphology was observed by transmission electron microscope. The endocytosis and exocytosis of the micelles at pH 7.4 and pH 6.0, as well as the effect of FcRn on the endocytosis, were investigated by flow cytometry. The results showed that the size of micelles was about 30 nm, which was not affected by FcBP decoration. We found that pH and ligand density could both influence the endocytosis. The uptake of active micelles was higher at pH 6.0 than at pH 7.4, and an optimal ligand density of endocytosis was appeared in both pH environment. Then we proved that FcBP decorated micelles could be endocytosed at pH 6.0 and exocytosed at pH 7.4, and the exocytosis process was also related to ligand density. Micelles with 10% ligand density had the largest exocytosis, showing the potentiality to deliver drugs through the intestinal epithelial. In addition, the competitive inhibition experiments illustrated that the interaction between FcRn and FcBP were essential to endocytosis. The results will enhance the understanding on the FcBP decorated PEG-PCL micelles for transmemberane drug delivery.

LEA (late embryogenesis abundant) proteins that are highly hydrophilic and thermally stable play a role in plant defense. The full-length cDNA of DoLEA2 was cloned by rapid amplification of cDNA ends (RACE) from Dendrobium officinale (GenBank number:KY626329). The cDNA is 1 224 bp and encodes 313 amino acids. The deduced DoLEA2 protein contained LEA_2 and WHy domains. Multiple sequence alignment revealed that DoLEA2 shared a high homology with other species. Phylogenetic tree showed that DoLEA2 belonged to the monocotyledon and its closest relative was P. aphrodite. DoLEA2 was differentially expressed in the different organ. The expression was most abundant in the leaves, followed by that of the roots and stem. DoLEA2 could express in Escherichia coli BL21 (DE3), and the best induction conditions were 0.5 mmol·L^-1 IPTG at 37℃ for 4 h. The growth curves of E. coli BL21 (DE3) showed that the recombinant DoLEA2 protein improved tolerate against salt stress over the control. This study represents the first time of cloning and identification of the function of LEA2 in D. officinale. The result sets up an important foundation for the molecular mechanism of stress resistance in Dendrobium officinale.