L-Proline-m-bis (2-chloroethyl) amino-L-phenylalanyl-L-norvaline ethyl ester hydrochloride (MF13) is a new anticancer tripeptide. Our previous study in vitro and in vivo showed that MF13 had anti-proliferative activities in a panel of human hepatocellular carcinoma (HCC) cell lines from different origin. In the present study, we focused on the inhibition effect on HCC of MF13 combined with other anti-cancer drugs. The results of combination chemotherapy in vitro indicated that the combination of MF13 with mitomycin C (MMC) at appropriate concentrations led to a synergistic effect; however, the combination of MF13 with vincristine (VCR) showed no synergistic effect. In the Bel-7402 tumor bearing nude mice, the antitumor effect of the groups of 2 mg·kg^-1 MF13 + 2 mg·kg^-1 MMC or 2 mg·kg^-1 MF13 + 50 mg·kg^-1 cyclophosphamide (CTX) exhibited synergistic anticancer efficacies while the group of 2 mg·kg^-1 MF13 + 0.3 mg·kg^-1 VCR did not have the same effect. Based on our data, we believe that MF13 can be considered as a potential agent against human hepatocellular carcinoma no matter how treated, alone or combined with other drugs.

The mesoporous silica nanoparticles (MSN) in different pore size and sirolimus (SRL) loaded self-microemulsifying drug delivery system (SMEDDS) were prepared. The results in morphology were collected by scanning electron microscope, transmission electron microscope, small-angle X-ray diffraction, and N₂ adsorption-desorption. The results showed that the prepared MSN has ordered nanochannels with a pore size of 6.3, 8.1, 10.8 nm, respectively. The particle size of SRL-SMEDDS were measured by particle sizing system, which was 20.6±1.3 nm. The stirring method was developed to prepare SRL-SMEDDS-MSN. It was found that the optimal ratio of SRL-SMEDDS to MSN was 2:1, while the drug loading rate was near 0.83%, and the flow properties of SRL-SMEDDS-MSN were of good condition. The differential scanning calorimetry results proving a molecular or amorphous dispersed state of SRL in MSN while the suspension experiment has shown great reconstitution properties of SRL-SMEDDS-MSN. There is no significant influence on maximum drug release rate of different pore size of SRL-SMEDDS-MSN in 250 mL water within 2 h, while the results of the first 40 min have an obvious difference. Above all, MSN might provide a new strategy for the solidification of SMEDDS.

The paper was aimed to investigate the association of VDR polymorphisms with tacrolimus (FK506) concentration in Chinese renal transplant recipients. A total of 114 renal transplant recipients receiving tacrolimus were genotyped for VDR rs1540339 and rs2853559 by Agena Bioscience MassARRAY^® system and CYP3A5*3 by PCR-RFLP method. Trough concentrations of tacrolimus on day 7 after renal transplantation were collected from clinical data. Statistical analysis was performed with Spearman's correlation, Mann-Whitney U test and Kruskal-Wallis H test. The dose-adjusted concentration of tacrolimus in VDR rs2853559 GA and GG carriers were considerably higher than that of AA carriers. After stratification by CYP3A5*3 genotypes, VDR rs2853559 GA and GG carriers had a higher dose-adjusted tacrolimus concentration than that in AA carriers in CYP3A5 nonexpresser. CYP3A5*3 and VDR rs2853559 explained 45.6% variability of tacrolimus C₀/D. In CYP3A5 non-expressers, VDR rs2853559 explained 14.4% variability of tacrolimus C₀/D. The results illustrated that VDR rs2853559 polymorphisms was associated with tacrolimus concentrations, and the determination of this SNP may be useful for individualized medicine of tacrolimus.

Complement activation-related pseudo-allergic reactions (CARPA) may represent 77% of all immune-mediated immediate hypersensitivity reactions. Because of the universality of the CARPA response and correlation between it and drug properties, complement activity tests are recommended as one of the tests for immunotoxicity and bioequivalence of drugs. However, in-vivo tests of complement activation are complicated, and the immunological differences between different individuals and between human and animal, making it very necessary to establish a standard and sample evaluation model for testing the effects of drugs on complement activity. In this study, the standard reaction serum was prepared by pooling sera collected from 40 healthy blood donors; a standard positive control was prepared by incubation with a heat-agglutinated IgG and zymosan A; SC5b-9, C5a, C4d and Bb were chosen as the test targets and evaluation criteria of the results was defined, all of these constituted the in-vitro model. By using this in-vitro model, the immunological toxicity of the different prescription of antifungal drug amphotericin B, and voriconazole for injection, and the bioequivalence of amphotericin B liposome formulations were studied.

Parkinson's disease (PD) is the most prevalent neurodegenerative disorder, with several risk factors contributing to the onset, such as aging, genetics, oxidative stress and neuroinflammation. There are several PD animals that mimics different risk factor. α-Synuclein mutation mice and systemic lipopolysaccharide (LPS) injection mice are two kinds of most common animal models that replicate genetic mutation and neuroinflammation, respectively. However, in these two animal models, the pathogenesis occurred after a long period of stimulation. In the present study, four-month-old α-synucleintransgenic mice (A53T) were intraperitoneally injected with LPS once a week for continuous 8 weeks to simulate the inflammatory response. The behavioral results showed that the time of mice staying on the rod and the performance score were markedly decreased, indicating motor dysfunction. Dopaminergic neuronal function also decreased. It was noted that the movement dysfunction and pathological changes were aggravated in LPS plus α-synuclein challenged mice compared with LPS or α-synuclein stimulated alone, suggesting that the double attack had synergistic effects. Mechanistic study demonstrated that LPS and α-synuclein combined challenge led to obvious neuroinflammatory response and apoptosis, which might contribute to motor and dopaminergic neuronal dysfunction. In addition, differential proteomic study showed that the expression of CD99L2 and COX7RP significantly increased in the midbrain of LPS plus α-synuclein challenged mice, which were closely related to inflammation and apoptosis, and might be involved in the pathogenesis of PD. In conclusion, the present study demonstrated that LPS could potentiate dopaminergic neuronal function in α-synuclein transgenic mice, which might be an ideal method to develop PD animal model.

Rhizoma Dioscoreae Bulbiferae is a traditional Chinese medicine with hepatotoxicity, but the metabolic profile of fatty acids has not been identified in the rats with liver injury. In this project, a gas chromatography-mass spectrometry method was applied to simultaneous quantification of 16 non-esterified fatty acids (NEFA) and esterified fatty acids (EFA) in the serum of control, ethanol extraction of Rhizoma Dioscoreae Bulbiferae (ethanol extraction, ET) and diosbulbin B (DB)-treated rats. Meanwhile, the change of fatty acid metabolic profile of liver injured rats was analyzed by principal component analysis (PCA) and partial least squares-discriminant analysis (PLS-DA). The results of NEFA concentration indicated that the serum concen-trations of palmitic acid (C16:0), stearic acid (C18:0), palmitoleic acid (C16:1n7), oleic acid (C18:1n9), vaccenic acid (C18:1n7), linoleic acid (C18:2n6), linolenic acid (C18:3n3), eicosatrienoic acid (C20:3n6), arachidonic acid (C20:4n6) and docosahexaenoic acid (C22:6n3) in DB-treated rats decreased significantly, while that of C18:2n6 and C20:3n6 obviously increased and that of C20:4n6 and C22:6n3 noticeably dropped in ET-treated rats when compared with control. Furthermore, the results of EFA concentration illus-trated that the serum concentrations of C16:0, C18:0, C20:4n6, C22:6n3 and eicosapentaenoic acid (C20:5n3) in two toxic groups were remarkably decreased when compared with control. The fatty acid meta-bolic profiles of the two toxic groups exhibited significant difference from the normal levels, and the degree of deviation of ET group was higher than that of DB group. More importantly, the results of PLS-DA showed that C20:4n6 and C22:6n3 were important indicators of the hepatotoxicity induced by ET and DB, and the serum concentrations of the two fatty acids had good correlation with the levels of alanine aminotransferase, aspartate aminotransferase and total bilirubin using Pearson's correlation analysis and canonical correlation analysis (CCA). Therefore, C20:4n6 and C22:6n3 were identified as potential biomarkers of ET and DB-induced liver injury. The project can provide a foundation for furture investigation of molecular mechanism of hepato-toxicity caused by Rhizoma Dioscoreae Bulbiferae.

Acting as the key step of various cell signaling pathways, protein-protein interaction (PPIs) plays a significant role in the regulation of biological functions. Many of the proteins involved are potential drug targets, therefore manipulation of PPI would greatly a great interest in physiology and pharmacology. Most PPIs could not be directly targeted by small molecule drugs due to their flat and shallow interaction surfaces. Selective regulator may be obtained by extraction and chemical synthesis helical peptide that forms folding substructure scaffold in PPI. However, most peptides when separated from its protein progenitor are not able to maintain its biological active helical structure, but tend to form random coil conformation, which is vulnerable to enzyme and suffer low potency and drugability. By modification through all-hydrocarbon bridged cyclic peptide designating 'stapled peptide', it turned out to be the most effective and directive methodology to solve the drawbacks. Stapled peptide not only can boost its potency, but also its biostability and cell permeability. These significant advantages make peptide stapling an important way of modification, which may definitely generate more and more peptide drug candidates targeting PPIs in the foreseen future. In this paper, chemistry and bioactivity of the stapled peptides reported up-to-date are systematically reviewed and discussed.

A series of novel benzimidazole and benzothiazole derivatives were designed and synthesized as inhibitors of SIRT1-SIRT3. The target compounds were synthesized from potassium O-ethyldithiocarbonate through a three-step route. The structures of the obtained compounds were elucidated by ¹H NMR and HR-MS. Of all compounds, six showed potent SIRT2-inhibitory activities with IC₅₀ values ranging from 2.8 to 21.2 μmol·L^-1. Among them, compound 10c displayed the most potent SIRT2-inhibitory activities (IC₅₀ = 2.8 μmol·L^-1), with more than 35-fold selectivity over SIRT1 and SIRT3 (IC₅₀ > 100 μmol·L^-1).

The study was aimed to investigate the correlation between the immunocompetence and the fingerprints of supernatant extracts from Radix Astragali by multi-index integrated evaluation, and reveal the material basis of Radix Astragali improving the immunological function. After oral administration 10 batch of supernatant extracts from Radix Astragali on immunosuppressed mice, phagocytic index, delayed type hypersensitivity (DTH) degree, organs index and the level of cytokines IFN-γ, IL-4 (ELISAs) were measured, and each common peak from HPLC-DAD/HPLC-ELSD fingerprints were correlated with the above data. A number of components in supernatant extracts of Radix Astragali display a substantial correlation with the immunocompetence. Supernatant extracts of Radix Astragali can significantly enhance immunological function on mice, which is related to various components in Radix Astragali.

To develop a taste-masking oral preparation of azithromycin for pediatrics, the reversed lipid nano-micelle techniques were used to mask the bitterness of azithromycin. Dry emulsion (DE) for taste-masking was prepared by solidifying the reversed-micelle oil solution containing azithromycin. Colloidal silicon dioxide was used as absorbent and solid carrier. Solidification was confirmed through dying test and observed by scanning electron microscope (SEM). The DE formulation was characterized by X-ray powder diffraction and SEM in order to investigate the crystal state of drug. Reconstitution emulsion droplet size and morphology were also determined using Nano ZS90 Zetasizer and transmission electron microscopy (TEM). The taste testing was performed in two different ways, namely, human taste panel test and the measurement of the amount of drug released in simulated oral cavity condition. The intestinal mucosal irritation test of DE formulation was also investigated in rats in comparison with commercial product (Zithromax). The optimal taste-masking formulation of azithromycin can be re-dispersed immediately with mean diameter of 530.1 nm after agitation in water. The results of taste testing showed that the bitterness of azithromycin was successfully masked by DE formulation similar with Zithromax at the same dose, moreover reduced intestinal irritation compared to Zithromax. These results indicate that the DE formulation for taste-masking of azithromycin is promising and valuable in the future development of azithromycin for pediatrics.