Compound Yizhihao, consists of Radix isatidis, Folium isatidis, Artemisia rupestris, has a significant therapeutic effect on the treatment of influenza and fever. However, the mechanism of its action is still unclear. In this investigation, we collected the key target molecule of influenza disease and the chemical constituents of Compound Yizhihao, and developed Naïve Bayesian classification models based on the input molecular fingerprints and molecule descriptors. The built models were further applied to construct classifiers for predicting the effective constituents. We used the professional network-building software to build the constituent-target network and target-pathway network, which revealed the network pharmacology of the effective constituents in Compound Yizhihao. It will contribute to the further research of mechanism of Compound Yizhihao.

The study was aimed to establish a liquid chromatography-tandem mass spectrometric method for the determination of the duloxetine concentration in rat plasma, and compare the pharmacokinetics in normal and diabetes mellitus rat models. Diazepam was used as an internal standard. The separation was achieved on a Waters Xterra^® RP18 column (100 mm × 4.6 mm, 3.5 μm) with a mobile phase consisting of methanol-0.3% formic acid containing 5 mmol·L^-1 ammonium acetate (75:25) at the flow rate of 0.6 mL·min^-1. Electrospray ionization source was applied and operated in the positive multiple reaction monitoring mode. A good linearity of duloxetine was obtained in the concentration range of 10-5 000 ng·mL^-1. The rat models of diabetes mellitus were established by intraperitoneal injection of streptozotocin. The same dose of duloxetine (40 mg·kg^-1) was given by intragastric administration to the normal and diabetic rats. Blood samples were collected from the orbital venous plexus to determinate duloxetine concentration in the plasma. The pharmacokinetic parameters were calculated by DAS software. Statistical analysis was performed by SPSS software. The major pharma-cokinetic parameters of diabetes group were as follows: C_max was 1 185 ± 190.0 ng·mL^-1; AUC_0-∞ was 8 398 ± 1 835 ng·mL^-1·h; t_max was 1.6 ± 0.4 h; t_1/2z was 3.6 ± 0.9 h. The major pharmacokinetic parameters of normal group were as follows: C_max was 368.1 ± 40.7 ng·mL^-1; AUC_0-∞ was 4145 ± 640.1 ng·mL^-1·h; t_max was 1.6 ± 0.3 h; t_1/2z was 4.1 ± 0.8 h. The results of pharmacokinetic experiments suggest that the exposure amount of duloxetine in diabetic rats is twice higher than that in normal rats.

Lepidium apetalum was used as an experimental material in this study. By analyzing the tran-scriptome data of L. apetalum and application of the specific primers, cDNA of cinnamate-4-hydroxylase (C4H) gene was isolated from L. apetalum and named as LaC4H (GenBank accession No. KX064050). Meanwhile, the bioinformatic analysis, prokaryotic expression, purification, tissue-specific expression analysis and expres-sion analysis after methyl jasmonate (MeJA) treatment were carried out. The results indicated that: ① The open reading frame (ORF) of LaC4H was 1 518 bp, which encoded a protein of 505 amino acid residues, with a predicted molecular mass of 57.73 kD. ② Bioinformatic analysis showed that LaC4H protein contained the conserved sequences of cytochrome P450 (CYP450) and 5 substrate recognition sites (SRSs) of CYP73A1, therefore LaC4H protein was a member of CYP450 superfamily. The phylogenetic analysis indicated that LaC4H protein showed the highest homology with C4H protein from cruciferous plants (such as AtC4H from Arabidopsis thaliana). ③ Through the construction of the prokaryotic expression vector pET-32a-LaC4H, the recombinant LaC4H protein was successfully expressed in E. coli BL21 (DE3) cells and the recombinant LaC4H protein was purified by Ni²⁺ affinity chromatography. ④ Real-time PCR analysis indicated that LaC4H was expressed in a high transcript level in stems, lower levels in leaves and flowers, the lowest level in roots. After MeJA treatment, the expression level of LaC4H in leaves was increased significantly to reach the highest level at 48 h. Furthermore, the expression levels of LaC4H were positively correlated with the flavonoids contents in leaves. The results of this study provide the fundamental information on LaC4H gene in the flavonoids biosyn-thesis pathway of L. apetalum.

To develop a cell-penetrating peptide with high membrane penetrating ability and effective antitumor activity, we designed and synthesized an analogue of penetratin, [Cys-CPT^{2, 9}] penetratin, by substitution of Gln² and Asn⁹ with Cys-CPT. We investigated the transmembrane activity and antitumor activity of [Cys-CPT^{2, 9}] penetratin. The fluorescence intensity of [Cys-CPT^{2, 9}] penetratin in HeLa cells was observed by laser confocal microscopy and flow cytometry, and the cell uptake mechanism of [Cys-CPT^{2, 9}] penetratin was evaluated by using different endocytic inhibitors, finally the anti-tumor activity of [Cys-CPT^{2, 9}] penetratin was tested by MTT assay. The results showed that the membrane activity of [Cys-CPT^{2, 9}] penetratin was significantly enhanced in laser confocal microscopy and flow cytometry assay, and the intracellular fluorescence intensity was 5 times higher than penetratin. The cell uptake mechanism study of [Cys-CPT^{2, 9}] penetratin indicated that it mainly entered the cell through the clathrin and endocytosis. Moreover, [Cys-CPT^{2, 9}] penetratin exhibited anti-tumor activity against HeLa cells, and its inhibitory effect on cancer cells was stronger than CPT. [Cys-CPT^{2, 9}] penetratin is a new cell-penetrating peptide with high translocation ability, and has anti-tumor activity against HeLa cells.

In our study of the chemical constituents of the dried mature fruits of Arctium lappa L., ten compounds were isolated by various chromatography methods and preparative HPLC. Their structures were elucidated as (7R, 8R)-4, 7, 9, 9'-tetrahydroxy-3, 3'-dimethoxy-8-4'-oxyneolign-7'-ene-9'-O-β-D-glucopyranoside (1), (7R, 8R)-4, 7, 9, 9'-tetrahydroxy-3, 3'-dimethoxy-8-O-4'-neolignan-9'-O-β-D-glucopyranoside (2), (7R, 8R)-4, 7, 9, 9'-tertahydroxy-3, 3'-dimethoxy-8-4'-oxyneolignan (3), (7S, 8R)-dihydrodehydrodiconiferylalcohol-4-O-β-D-glucopyranoside (4), (7S, 8R, 7'R, 8'R)-pinoresinol-4, 4'-di-O-β-D-glucopyranoside (5), (8S, 7'S, 8'R)-4, 4', 9'-trihydroxy-3, 3'-dimethoxy-7', 9-epoxylignan-7-oxo-4'-O-β-D-glucopyranoside (6), 2-methoxy-4-hydroxyphenol-1-O-β-D-xylopyranosyl-(1→6)-O-β-D-glucopyranoside (7), 3-methoxy-4-hydroxyphenol-1-O-β-D-xylopyranosyl-(1→6)-O-β-D-glucopyranoside (8), 4-hydroxy-3-methoxybenzylalcohol-4-O-β-D-xylopyranosyl-(1→6)-O-β-D-glucopyranoside (9) and 2-phenethyl β-primeveroside (10) by their spectroscopic data (IR, UV, CD, MS, HR-ESI-MS, and 1D and 2D NMR) and comparison to literature data. Compound 1 is a new 8-O-4'-neolignan. Compounds 2-10 were isolated from the dried mature fruits of Arctium lappa L. for the first time.

This study was aimed to build a new photo-sensitive co-delivery liposomes which combine photodynamic therapy with chemotherapy to reverse drug resistance of breast cancer. Photodynamic photosen-sitizer chlorin e6 trimethyl ester (Ce6tM) and chemotherapeutic drug doxorubicin hydrochloride (DOX) were loaded into the liposomes (liposomes loaded with Ce6tM and DOX, CDL) by thin-film hydration extrusion and ammonium sulfate active loading methods. CDL was characterized with cryo-transmission electron microscopy (Cryo-TEM), dynamic light scattering particle size, zeta potentials and photo-sensitive DOX release behaviors in vitro. CDL cytotoxicity, singlet oxygen production, DOX accumulation, intracellular ATP level and cell cycle analysis in MCF7/ADR cells were evaluated. Finally, the tissue distribution of DOX and antitumor effects of CDL in BALB/c-nu nude mice bearing MCF7/ADR tumor were investigated. The results showed that the particle size of obtained CDL was 90.7 ± 1.1 nm and distributed uniformly. CDL possessed outstanding properties of photo-sensitive drug release profile. The accumulated release of DOX reached (96.52 ± 0.11)% in 2 min under 671 nm laser irradiation (2 W·cm^-2). Interestingly, DOX in CDL could maintain rapid release after 671 nm laser irradiation with low power and short time (15 s, 0.25 W·cm^-2). This phenomenon was caused by oxidation of unsaturated phospholipids in CDL under 671 nm laser irradiation and had nothing to do with the slightly elevated temperature. Photo-sensitive drug release behavior contributed to increased DOX accumulation in MCF7/ADR cells. The half inhibition concentration (IC₅₀) of DOX in CDL laser group in MCF7/ADR cells was decreased by 601.9-fold compared with no laser group, which could be related to increased accumulation of DOX, decreased ATP levels and cell cycle arrest in MCF7/ADR cells. With the help of CDL, DOX accumulation in tumor was increased and in cardiac toxicity was reduced in vivo. CDL laser group showed a good anti-tumor effect. The tumor inhibition rate was (94.7 ± 6.2)%. These results suggest that CDL has a promising potential in reversing drug resistance of breast cancer.

The appropriate regulation of intracellular bioenergy and nutrient metabolism is a basic requirement for proper function and survival of pancreatic beta cells, where mitochondria-endoplasmic reticulum (ER)-associations play crucial roles. Mitochondria are changed dynamically according to intracellular energy and nutrients, which provides material foundation for energy homeostasis; while ER regulates metabolic enzymes and protein synthesis in different pathways. This review sheds light upon the development of mitochondria-ER associations and its role in the regulation of insulin secretion in pancreatic beta cell. The impact on beta cell viability is discussed. Interruption of calcium and redox oxidative species results in reduction of glucose-stimulated insulin secretion, while intracellular calcium levels could be partial altered by depleting calcium from the ER. Given the tight link between ER and mitochondria, the association are crucial to the homeostasis and are an indicator of overall beta cell status, with a potential as a novel drug target for treatment of type 2 diabetes mellitus.

Serotonin (5-hydroxytryptamine, 5-HT) is an endogenous molecule playing key roles in life activity. In the treatment of metabolic diseases such as obesity, the traditional paradigm of appetite suppression by serotonin in the central nervous system has some limitations. In recent years, great progress has been made in the study of the regulation of energy metabolism by peripheral serotonin system. Recent studies revealed that peripheral serotonin plays an important role in regulation of adipogenesis and energy expenditure in adipose tissues, insulin secretion in pancreatic β-cell and glycogen synthesis in liver, etc. This review summarizes the recent advances in the function of serotonin in the peripheral organs of energy metabolism. We propose that peripheral serotonin system may serve as an attractive new therapeutic target for the treatment of metabolic diseases in the near future.

Lymphoid-specific tyrosine phosphatase (LYP) is a phosphatase that is encoded by protein tyrosine phosphatase non-receptor type 22 and is mainly distributed in lymphoid. In psychological condition, LYP inhibits T-cell receptor (TCR) signaling in association with C-terminal kinase (CSK). While in pathological condition, mutant LYP dissociates with CSK, which augments the inhibition of TCR signaling and leads to autoimmune diseases. Consequently, LYP is now considered as a new target of type Ⅰ diabetes, rheumatic arthritis and Graves disease and some other autoimmune disorders. This review mainly focuses on the development of LYP inhibitors in their structures and activities.

Transient receptor potential (TRP) channels are non-selective and cation-permeable channels in the cell membrane, widely distributed in tissues and organs of human body. As biosensors, TRP channels can regulate the functions of vision, hearing, taste, pain, and touch, etc. So far, more than 100 different kinds of natural modulators targeting TRP channels have been identified from 70 species of plants or animals. In this review article, we attempt to summarize the effect of known natural active compounds on TRP channels with focuses on their sources, structures, action features and mechanisms. Hopefully this review can provide some useful information that can facilitate discovery of more specific natural modulators, and development of innovative therapeutic drugs targeting TRP channels.