Sanhuang Xiexin Decoction (SXD) is commonly used to treat type 2 diabetes mellitus (T2DM) in clinical practice of traditional Chinese medicine (TCM). In order to elucidate the specific analysis mechanisms of SXD for T2DM, the method of network pharmacology was applied to this article. First, the effective ingredients of SXD were obtained and their targets were identified based on the TCMSP database. The T2DM-related targets screened from the GEO database were also collected by comparing the differential expressed genes between T2DM patients and healthy individuals. Then, the common targets in SXD-treated T2DM were obtained by intersecting the putative targets of SXD and the differential expressed genes of T2DM. And the protein–protein interaction (PPI) network was established using the above common targets to screen key genes through protein interactions. Meanwhile, these common targets were used for GO and KEGG analyses to further elucidate how they exert antidiabetic effects. Finally, a gene pathway network was established to capture the core one in common targets enriched in the major pathways to further illustrate the role of specific genes. Based on the data obtained, a total of 67 active compounds and 906 targets of SXD were identified. Four thousand one hundred and seventy-six differentially expressed genes with a P value < 0.005 and ∣log2fold change∣>0.5 were determined between T2DM patients and control groups. After further screening, thirty-seven common targets related to T2DM in SXD were finally identified. Through protein interactions, the top 5 genes (YWHAZ, HNRNPA1, HSPA8, HSP90AA1, and HSPA5) were identified. It was found that the functional annotations of target genes were associated with oxygen levels, protein kinase regulator, mitochondria, and so on. The top 20 pathways including the PI3K-Akt signaling pathway, cancers, HIF-1 signaling pathway, and JAK-STAT signaling pathway were significantly enriched. CDKN1A was shown to be the core gene in the gene-pathway network, and other several genes such as CCND1, ERBB2, RAF1, EGF, and VEGFA were the key genes for SXD against T2DM. Based on the network pharmacology approach, we identified key genes and pathways related to the prognosis and pathogenesis of T2DM and also provided a feasible method for further studying the chemical basis and pharmacology of SXD.

Muscarinic acetylcholine receptors (mAChRs) have five subtypes and play crucial roles in various physiological functions and pathophysiological processes. Poor subtype specificity of mAChR modulators has been an obstacle to discover new therapeutic agents. Muscarinic toxin 7 (MT7) is a natural peptide toxin with high selectivity for the M1 receptor. With three to five residues substituted, M3, M4, and M5 receptor mutants could bind to MT7 at nanomolar concentration as the M1 receptor. However, the structural mechanisms explaining MT7–mAChRs binding are still largely unknown. In this study, we constructed 10 complex models of MT7 and each mAChR subtype or its mutant, performed molecular dynamics simulations, and calculated the binding energies to investigate the mechanisms. Our results suggested that the structural determinants for the interactions on mAChRs were composed of some critical residues located separately in the extracellular loops of mAChRs, such as Glu4.56, Leu4.60, Glu/Gln4.63, Tyr4.65, Glu/Asp6.67, and Trp7.35. The subtype specificity of MT7 was attributed to the non‐conserved residues at positions 4.56 and 6.67. These structural mechanisms could facilitate the discovery of novel mAChR modulators with high subtype specificity and enhance the understanding of the interactions between ligands and G‐protein‐coupled receptors. Copyright © 2015 John Wiley & Sons, Ltd.

Citalopram is a selective serotonin reuptake inhibitor (SSRI) mainly prescribed to treat major depression.

Background Citalopram is a selective serotonin reuptake inhibitor (SSRI) mainly prescribed to treat major depression.Objective The aim of this study was to compare the pharmacokinetic characteristics of a new and a branded citalopram 20 mg formulation to support the marketing authorization of the test formulation in China.Methods A single-dose, open-label, randomized-sequence, two-period crossover design was used in this study. Healthy Chinese male cytochrome P450 (CYP) 2C19 extensive metabolizers, aged 18-40 years, were eligible to participate. CYP2C19 poor metabolizers were excluded, based on genotyping of genomic DNA from blood samples. Twenty-four subjects were randomly assigned to receive the test formulation followed by the reference formulation, and then vice versa. A 2-week washout occurred between study periods. Blood samples were collected for up to 144 h post-dose. Quantification was carried out using a validated high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method. Pharmacokinetic parameters were calculated and analysed statistically. The two formulations were considered pharmacokinetically equivalent if the 90 % confidence intervals (CIs) of the log-transformed ratios (test/reference) of the maximum plasma concentration (C-max), area under the plasma concentration-time curve from time zero to the last measurable concentration (AUC(last)), and area under the plasma concentration-time curve from time zero to infinity (AUC(infinity)) were within the predetermined acceptance range (70-143 % for C-max; 80-125 % for AUC(last) and AUC(infinity)) according to China State Food and Drug Administration bioequivalence guidelines. Tolerability was monitored by clinical assessment, vital signs, laboratory analysis and interviews with participants about adverse events.Results A total of 24 participants, with a mean (SD) age of 26 (3) years (range 22-32 years), body weight of 65.2 (5.0) kg (range 53-73 kg), and height of 172.7 (4.9) cm (range 159-182 cm), were enrolled in this study. Both formulations showed similar pharmacokinetic profiles. Mean (SD) AUC(last), AUC(infinity) and C-max were 1436 (341) ng . h/mL, 1595 (381) ng . h/mL and 32.3 (5.9) ng/mL, respectively, for the test formulation, and 1444 (388) ng . h/mL, 1648 (504) ng . h/mL, 33.1 (7.4) ng/mL, respectively, for the reference formulation. Median (range) time to reach C-max (t(max)) was 2 (1-12) hours (test) and 3 (1-6) hours (reference). The 90 % CIs of the treatment ratios for the ln-transformed values of C-max, AUC(last) and AUC(infinity) were 92.5-103.6, 95.2-100.6 and 96.4-105.4, respectively. No significant difference was found between treatments with regard to pharmacokinetic parameters. Fifteen adverse events were reported during the study but none were considered serious.Conclusion This single-dose study found that the test and reference citalopram 20 mg tablets met the regulatory criteria for assuming bioequivalence in the selected healthy Chinese male subjects. Both formulations were well tolerated.

Objective To explore the protective effect of active component of Eclipta on hepatic injury induced by Concanavalin A(ConA) in mice.Methods Mouse hepatic injury models were established by injection of ConA(15 mg/kg) via tail vein.The effect of Eclipta extracts on serum alanine aminotransferase(ALT) and liver histology in both normal mice and hepatic injury mice was examined.A pure compound was obtained by means of bioactivity-guided isolation,and IR, ~(13)C-NMR,~1H-NMR,~1H-~(13)C HMBC and ~1'H-~(13)C HMQC were used for structural identification.Results Eclipta extracts decreased serum ALT in hepatic injury mice induced by ConA,and demonstrated anti-apoptosis effect induced by ConA in hepatocytes.Wedelolactone,a coumarin,was isolated from Eclipta,and its structure was identified on the basis of spectroscopic analysis.Wedelolactone inhibited ConA-induced T cell proliferation selectively,and completely antagonized the effect of ConA at the concentration of 10μg/mL.Conclusion Eclipta herb has hepatoprotective effect. Wedelolactone,the anti-inflammatory and anti-apoptosis active component of Eclipta,might serve as a lead compound for developing new antiinflammatory drugs.

A simple sensitive and robust method for simultaneous determination of citalopram and desmethylcitalopram was developed using liquid chromatography tandem mass spectrometry (LC–MS/MS). A 200 μL aliquot of plasma sample was employed and deproteinized with methanol and desipramine was used as the internal standard. After vortex mixing and centrifugation, the supernatant was diluted with water (1:1, v/v) and then directly injected to analysis. Analytes were separated by a Zorbax XDB C18 column with the mobile phase composed of acetonitrile and water (30:70, v/v) with 0.25% formic acid and monitored in MRM mode using a positive electrospray source with tandem mass spectrometry detection. The total run time was 3.5 min. The dynamic range was 0.2–100 ng/mL for citalopram and 0.25–50 ng/mL for desmethylcitalopram, respectively. Compared to the best existing literatures for plasma samples, the same LOQ for CIT (0.5 ng/mL) and lower LOQ for DCIT (0.25 vs 5 ng/mL) were reached, and less sample preparation steps and runtime (3.5 vs 10 min) were taken for our method. Accuracy and precision was lower than 8% and lower than 11.5% for either target. Validation results and its application to the analysis of plasma samples after oral administration of citalopram in healthy Chinese volunteers demonstrated the method was applicable to pharmacokinetic studies.

Objective To evaluate the strengthening and restoring effects and hepatoprotective effect of Eclipta extracts.Methods Sixty Kunming mice were randomly divided into control group,low-dose Eclipta group,high-dose Eclipta group,cyclophosphamide(CY) group,low-dose Eclipta CY group and high-dose Eclipta CY group(n=10).Eclipta 8 g·kg-1·d-1 or 4 g·kg-1·d-1 were administered by oral gavage for 7 d.Immunosuppression mouse models were established by intraperitoneal injection of 0.05 g/kg CY on the fourth day of administration.The effects of Eclipta extracts on thymus index,spleen index and carbon clearance index in both normal mice and immunosuppression mice induced by CY were tested,and those on proliferation of lymphocyte and T cells induced by ConA were examined.Based on these data,strengthening and restoring effects of Eclipta were observed.Besides,70 Kunming mice were randomly divided into control group,low-dose Eclipta group,high-dose Eclipta group,CCl4 group,DDB group,low-dose Eclipta CCl4 group and high-dose Eclipta CCl4 group.20 g·kg-1·d-1 and 10 g·kg-1·d-1 Eclipta were administered by oral gavage for 7 d.Hepatic injury mouse models were established by intraperitoneal injection of 10 mL/kg CCl4 2 h after the last administration.The effects of Eclipta extracts on serum ALT,MDA,TP and T-Bil levels in both normal mice and hepatic injury mice induced by CCl4 was tested,and those on the viability of the primary culture hepatocytes were determined.Based on these data,hepatoprotective effect of Eclipta was observed. Results Eclipta extracts increased thymus index and carbon clearance index in both normal mice and immunosuppression mice induced by CY,promoted proliferation of lymphocyte and proliferation of T cells induced by ConA,reduced serum ALT and MDA levels in hepatic injury mice,and improved the viability of the primary culture hepatocytes. Conclusion Eclipta herb has strengthening and restoring effects and hepatoprotective effect.

A new full-length cDNA encoding strictosidine synthase (designated as OjSTR, GenBank Accession No. 1087598), which catalyzes a committed step in camptothecin biosynthetic pathway, was isolated from young leaves of Ophiorrhiza japonica for the first time. OjSTR was 1,258 bp and contained a 1,062 bp open reading frame encoding a deduced protein of 353 amino acid residues. Sequence analyses showed that OjSTR had high homology with other STRs from some TIA-producing plants. Phylogenetic tree analysis showed that OjSTR had closest relationship with STR from O. pumila. Tissue expression pattern analysis revealed that OjSTR constitutively expressed in all the tested tissues at different levels, which was high in flower, moderate in leaf and root, low in stem. Expression profiles under plant defense signals such as methyl jasmonate and salicylic acid were investigated, and the results revealed that expression of OjSTR was all induced, implying that OjSTR was high elicitor responsive.

In this study, an efficient transformation system for the medicinal plant Anisodus acutangulus was successfully developed and optimized using Agrobacterium rhizogenes . Three bacterial strains, A4, R1601, and modified C58C1 and three explant types, leaf blade, petiole, and stem, were examined. The highest transformation efficiency of 94.44% was achieved using strain C58C1 with stem explants. Over 20 independent hairy root lines were successfully established with strain C58C1 using stem explants, all of which contained the ro/B and ro/C genes as confirmed by polymerase chain reaction (PCR). Out of four media compositions, the liquid 1/2 MS medium was found the most suitable for hairy root growth. The maximum biomass of one hairy root line increased up to 80 times in liquid 1/2 MS medium after a 30 day culture period. Different hairy root lines displayed a varied capacity for tropane alkaloid production and the best hairy root line (T4) from the C58C1-stem combination produced up to 10.21 mg/g (dw) of hyoscyamine, which was about 1.5-fold higher than in the wild type plants. To our knowledge, this is the first report to demonstrate the production of tropane alkaloids in hairy roots of A. acutangulus .

In this paper we report on the molecular cloning, sequencing and partially characterisation of a lectin from bulb of the Chinese medicinal plant Zephyranthes grandiflora. The full-length cDNA of Z. grandiflora bulb lectin (ZGBL) consisted of 986 bp and contained a 576 bp ORF encoding a 191 amino acid protein. Bioinformatics analysis results clearly indicate that ZGBL belongs to the monocot mannose-binding lectin family, which contains 3 putative mannose-binding sites per subunit. RT-PCR analysis results indicate that ZGBL is constitutively expressed in all the tested tissue types including root, bulb, leaf and flower. Interestingly, ZGBL is more closely related to the Orchidaceae rather than the Amaryllidaceae family on molecular evolution.

AIM: To investigation the anti-coxsackievirus B3 (CVB3m)effect of the ethyl acetate extract of Tian-hua-fen on HeLacells infected with CVB3m.METHODS: HeLa cells were infected with CVB3m and thecytopathic effects (CPE) were observed through light microscope and crystal violet staining on 96-well plate and A600 was detected using spectrophotometer. The protective effect of the extract to HeLa cells and the mechanism of the effect were also evaluated through the change of CPE and value of A600.RESULTS: The extract had some toxicity to HeLa cells at a higher concentration while had a marked inhibitory effect on cell pathological changes at a lower concentration.Consistent results were got through these two methods.We also investigated the mechanism of its anti-CVB3m effectand the results indicated that the extract represented an inhibitory effect through all the processes of CVB3m attachment, entry, biosynthesis and assemble in cells.CONCLUSION: The results demonstrate that the ethyl acetate extract of Tian-hua-fen has a significantprotectiveeffect on HeLa cells infected with CVB3m in a dose-dependent manner and this effect exists through the process of CVB3mattachment, entry, biosynthesis and assemble in cells,suggesting that the ethyl acetate extract of Tian-hua-fen can be developed as an anti-virus agent.

m5受体是M受体家族一个新成员 ,与其它亚型相比 ,由于缺乏高选择性配体和标记条件以及富含m5受体的组织来源 ,目前对其分子结构、结合特性、组织分布、生理功能等知之甚少。随着免疫沉淀、原位杂交和药理学标记方法的发展 ,现已证实m5受体主要分布于大脑皮质最外层 ,海马齿状回、CA1、CA2区 ,嗅结节 ,横核以及外周血淋巴细胞、单核细胞、睫状肌等部位 ,提示m5受体在调节大脑和外周组织胆碱能神经活动方面具有独特而重要的作用 ,有可能成为治疗帕金森病、阿尔采末病、局灶性脑缺血等疾病新的药物靶点。