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  • Yu-zhuo WU, Rui LI, Bo-lin QIU, Jing-fang ZHANG, Ling-yan WANG, Jun-gui DAI, Su-juan WANG, Hua-wei ZHANG, Sheng LIN
    Acta Pharmaceutica Sinica. 2017, 52(4): 598-602.

    A new phomapyrone derivative (1), and 9 known compounds were isolated from the ethyl acetate fraction of solid fermentation of Fusarium redolens, the endophytic fungus from Edgeworthia chrysantha, by using various isolation techniques such as Sephadex LH-20, MCI GEL and Semi pre-HPLC, etc. Their structures were identified by spectroscopic analysis, including MS, UV, CD, specific rotation, IR, 1D and 2D NMR, as (+)-7S-4-deoxy-9-hydroxyphomapyrone C (1), uracil (2), uridine (3), 2'-deoxyuridine (4), adenosine (5), 2'-deoxyadenosine (6), cordysinin B (7), ergosterol (8), ergosta-5α, 8β-epidioxy-6, 22-dien-3β-ol (9), and (22E, 24S)-3α-hydroxy-24-methylcholesta-5, 8, 22-trien-7-one (10).

  • Jing HU, Kun-ming QIN, Ting-ting ZHU, Xiao-li WANG, Meng-xue FAN, Bao-chang CAI
    Acta Pharmaceutica Sinica. 2017, 52(4): 603-608.

    In this study, we developed a qualitative analytical method based on liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (UHPLC-Q-TOF-MS/MS) for identification of multi-constituents of raw Fructus Arctii (RFA) and processed Fructus Arctii (PFA). We established a UHPLC-UV analytical method for simultaneously determining 6 major compounds in Fructus Arctii. UHPLC-Q-TOF-MS/MS qualitative analysis was performed under negative and positive ion modes and a total of 23 chemical compounds were identified. The analysis data were subjected to a principle component analysis with a t-test. Ten peaks were found to be the main difference (P < 0.05) between RFA and PFA. HPLC-UV quantitative method result showed the contents of 6 constituents were different between RFA and PFA. The results indicated that there was less arctiin, chlorogenic acid, isochlorogenic acid A in PFA than in RFA. However, there were higher levels of arctigenin, isochlorogenic acid B, isochlorogenic acid C in the PFA than RFA, which may be the main reason for different clinical efficacy of RFA and PFA.

  • Xuan QIN, Ang CHEN, Jian LU, Yuan-jin ZHANG, Ming-yao LIU, Xin WANG
    Acta Pharmaceutica Sinica. 2017, 52(4): 609-614.

    Parthenolide is a sesquiterpene lactone derived from the plant feverfew (Tanacetum parthenium) that possesses multiple anti-inflammatory and anti-cancer properties. A specific, sensitive, accurate and precise LC-MS/MS method was developed and validated in the quantitative analysis of parthenolide in rat plasma. A liquid-liquid extraction method was used to separate the analyte and the internal standard (IS, costunolide) from plasma. A water-methanol mobile phase system was utilized in the gradient chromatographic separation. The calibration curve with good linearity (r2 > 0.99) was established between 2 and 128 ng·mL-1 with accuracy and precision within acceptable limits at different QC levels. High extraction recovery was achieved for both parthenolide (89.55%-95.79%) and IS (96.87%). Based on this LC-MS/MS method, the plasma stability and pharmacokinetics of parthenolide were assessed in rats. Parthenolide was proved to be very unstable in rat plasma, and was distributed and eliminated quickly in vivo, with a half-life less than 90 min. A high dose of parthenolide (80 mg·kg-1) resulted in a very low initial concentration (138.86±21.07 ng·mL-1). The systemic exposure of parthenolide (area under the curve) increased disproportionally from 40 mg·kg-1 dose group to 80 mg·kg-1 dose group. The present study may provide helpful information for the development of parthenolide as a drug candidate.

  • Gui-zhen WANG, Xia HAO, Lei LIU, Zhen-yu LI, Xue-mei QIN, Guan-hua DU, Xiang ZHANG, Ke LI
    Acta Pharmaceutica Sinica. 2017, 52(4): 615-619.

    Due to the difference in structure and chemical properties of amino acids, it is difficult to determine amino acids with different properties by GC-MS. A method was established for the simultaneous determination of 20 amino acids by using ethyl chloroformate (ECF) as the derivatization reagent and adjusting pH 9-10 in the second derivatization in this study. The results showed that 20 amino acids were well separated and the compounds in the 2-20 μg·mL-1 concentrations were detected with a good linear response correlation coefficient r2 greater than 0.99. The results of precision and system adaptability show that RSD < 10%, the average recovery of samples in the 78.3%-109.9%. The results showed that the method was simple and easy to use, and the derivatization method could be carried out in aqueous solution. The derivatization product was stable with wide linear range, thus this method could be used for the determination of a variety of biological samples such as Astragalus mongolicus, mouse urine sample and mouse serum sample.

  • Xiao-han YANG, Guo-jie XU, Xin-yue ZHANG, Da LI, Hong-xia LI, Yu-feng SUN, Fan ZHANG, Chun-sheng LIU
    Acta Pharmaceutica Sinica. 2017, 52(4): 620-624.

    A rapid fluorescence polarization immunoassay (FPIA) has been developed for the determi-nation of aflatoxins in samples of naturally-contaminated herbal teas. The tracers were synthesized by chemical method and determined by thin layer chromatography (TLC) and mass spectroscopy (MS). Fluorescence polarization was evaluated by the detection of polarized light. The results showed that the limit of detection (LOD) of FPIA for aflatoxins was 20 ng·mL-1, the IC50 was 371.80 ng·mL-1, and the linear range of the developed FPIA was 92.76-252.32 ng·mL-1. Compared with conventional HPLC methods, the FPIA developed in this study has the advantages of short analysis time and low cost. This method may be suitable for high-throughput screening of aflatoxins in herbal teas.

  • Shu-jun LIANG, Sheng-yan ZHOU, Si-tong YANG, Dian-rong XIA, Yuan-feng WEI, Yuan GAO, Shuai QIAN, Jian-jun ZHANG
    Acta Pharmaceutica Sinica. 2017, 52(4): 625-633.

    The purpose of this study was to investigate the thermodynamics of naringenin (NAR)-isonicotinamide (INT) cocrystal (stoichiometric ratio, 1:2) formed in different solvents. The dissolution behavior of cocrystal was explored in the water. Solubility of NAR-INT cocrystals under various temperatures were measured, followed by fitting the complexation model to calculate the thermodynamic parameters solubility products (Ksp), complexation constants (K12) and Gibbs energy change (ΔG) of cocrystal during formation progress. Ternary phase diagrams (TPDs) of the NAR-INT-solvent systems under various temperatures were plotted. Based on the non-linear simulation, 1:2 complexation model was well fitted to the NAR-INT cocrystal formation in ethanol, isopropanol and ethyl acetate, while no complexation model was more suitable for that in methanol. The cocrystallization reaction was exothermic and spontaneous (ΔG < 0, ΔH < 0, ΔS < 0). Ksp increased while K12 decreased when increasing temperature, suggesting that the two components could cocrystallize more easily at the lower temperature. In comparison to TPDs in other solvents, the area of homogeneous liquid phase in ethyl acetate was the smallest, indicating the easiest formation of NAR-INT cocrystal in ethyl acetate. The current study provides a theoretical foundation for preparation and optimization of scale-up NAR-INT cocrystals.

  • Meng-meng ZHANG, Yu-ting YANG, Qian-wen YU, Qin HE
    Acta Pharmaceutica Sinica. 2017, 52(4): 634-640.

    In this study, we aim to develop a pH-sensitive transmembrane peptide TH (AGYLLGHINLHH LAHL (Aib) HHIL-Cys) modified liposome loaded with immunoadjuvant α-galactosylceramides (αGC-TH-Lip) and then investigate its effect on the immune function in tumor-bearing mice and its immune mechanism of action. The liposomes were prepared by membrane dispersion-probe ultrasound method and the size and zeta potential of αGC-TH-Lip were also characterized. The uptake of TH modified liposomes (TH-Lip) and polyethylene glycols modified liposomes (PEG-Lip) in DC2.4 cells in vitro were analyzed and the activation of natural killer T (NKT), natural killer (NK) and macrophages in tumor-bearing mice were also measured after systemic administrations of samples. Besides, the degree of maturation of dendritic cell (DC), the number of cytotoxic T lymphocyte (CTL) and the differentiation of helper T cell (Th) were determined. The results showed the particle size of αGC-TH-Lip was about 117.9 nm and the zeta potential was about-8.37 mV under the neutral condition (pH 7.4) and the αGC-TH-Lip had high serum stability in 50% fetal bovine serum. The uptake of TH-Lip in DC2.4 cells in vitro was 1.48 times higher than that of PEG-Lip. After systemic administrations of the samples, the numbers of NKT cells, NK cells and macrophages in tumor-bearing mice were (0.43±0.048)%, (12.80±0.50)% and (3.13±0.26)%, respectively, and the number of mature DCs and CTLs reached (2.30±0.22)% and (32.30±0.80)% separately, which was significantly different from the con-trol group. Finally, we discovered the αGC-TH-Lip had the strongest induction effect on the differentiation of Th1 cells, while barely promote the differentiation of Th2 cells. All the above results demonstrated that the αGC-TH-Lip can improve the immune the activity of mice, enhance the effect of α-galactosylceramide and promote the differentiation of lymphocytes toward the direction of cellular immunity, which consequently achieve a better anti-cancer immune activity.

  • Juan LIU, Tong CHEN, Yuan YUAN, Rui-feng JI, Juan GUO, Ying-ping WANG, Guo-liang WANG, Lu-qi HUANG
    Acta Pharmaceutica Sinica. 2017, 52(4): 641-646.

    Polar auxin transport gene PIN (PIN-FORMED) determines the concentration gradient of auxin and plays an important role in development and secondary metabolism of plants. This study was designed to analyze the bioinformatics and expression of the PIN genes in Panax ginseng to explore a novel way of breeding ginseng varieties. Heatmap and cluster analysis of PIN2, PIN3, PIN6 was performed in four-year-old Jilin ginseng. Sequence homology alignment, RT-PCR amplification, sequencing and bioinformatics analysis were used to identify three PIN family genes PgPIN2, PgPIN3 and PgPIN6 in P. ginseng. PIN expression in ginseng adventitious root and culture seedling was analyzed with qRT-PCR technique. Results suggested that in ginseng adventitious root tip, PgPIN3 and PgPIN6 exhibited a high level of expression; in ginseng culture seedling root, PgPIN2 showed a high level of expression; in four-year-old Jilin ginseng at the fruit ripening stage, PgPIN2 and PgPIN6 were highly expressed in root and rhizome, while PgPIN3 had a high level in ginseng leaf, fruit and root. Tissue specific expression profile showed that PgPIN2 and PgPIN6 probably were involved in the development and tropism growth in ginseng roots, while PIN3 might be in relation to the growth and development of the aerial part of plants.

  • Chao XIONG, Jing-jian LI, Wei SUN, He-gang LIU, Lan WU, Di LIU, Zhi-gang HU, Shi-lin CHEN
    Acta Pharmaceutica Sinica. 2017, 52(4): 647-652.

    Persicae semen has been used for years as a traditional Chinese medicine to treat diseases. Because of their similar morphologies, Persicae semen was commonly inadvertently mixed with Armeniacae semen amarum (a toxic herbal seed). Development of a reliable method for discriminating Persicae semen from its adulterant is necessary to reduce confusion for the drug safety in clinical practices. This study evaluates the efficiency of high-resolution melting (HRM) combined with internal transcribed spacer 2 (ITS2) to analyze Persicae semen. Our findings show that HRM allows not only the identification of adulteration but also the quantification of the most common admixture. HRM sensitivity in adulterant detection was assessed through the analysis of mixing samples with different proportions of Prunus persica and Prunus armeniaca control. The results are presented as a linear regression with r of 0.96 and imply the capability of the method to detect adulteration. In particular, HRM detected seeds of Prunus persica in Prunus armeniaca at concentrations as low as 1%, and commercial products labeled as 'Persicae semen' were purchased from markets and could rapid authenticated by HRM analyses. This study is significant in the verification of the authenticity in the quality control of herbal medicine. In the near future, it is promising to be the main trend for identifying traditional Chinese medicinal materials.

  • Ying PENG, Fei MA, Ting-ting XU, Shan WANG, Long-jian HUANG, Xiao-ying WANG, Yong-jun WANG, Xiao-liang WANG
    Acta Pharmaceutica Sinica. 2017, 52(3): 339-346.

    Translating of scientific advances into clinical practice is a major challenge in the stroke research field in the past decades. There were many reasons involved:animal models might not accurately capture all aspects of clinical stroke in humans, the blind and randomized design principle was not closely followed, the inclusion and exclusion criteria was not previously established, sample size was inadequate, endpoint was not scientific nor blindly assessed, inadequate reporting of data and statistical flaws. To bridge the gap between experimental and clinical research, international consortia have attempted to establish standardized guidelines for study design and data report, which include optimizing animal models as well as experimental design, using innovative approaches to assess endpoint, making raw data and negative results available, establishing prior registration mechanism, conducting multicenter preclinical randomized controlled trials (pRCTs), systematic reviews and meta-analysis of preclinical studies, evolving the original focus on neuroprotection into a broader consideration of the role of neurovascular unit and ischemic cascade.