Icariside II suppressed tumorigenesis by epigenetically regulating the circß-catenin-Wnt/ß-catenin axis in colorectal cancer.

Icariside II, a flavonol glycoside, one of the major components of Traditional Chinese Medicine Herba epimedii. In the present study, we found that Icariside II suppressed the proliferation of CRC by inducing cell cycle arrest and apoptosis in vitro and inhibited tumor growth in vivo. The further mechanism investigation showed that Icariside II suppressed the expression of ß-catenin and led to the functional inactivation of Wnt/ß-catenin signaling. Circß-catenin was considered as a promising candidate for mediating the tumorigenesis and the activation of Wnt/ß-catenin signaling in CRC cells. Furthermore, Icariside II has been proven to suppress the biogenesis of circß-catenin via epigenetically targeting DNA methyltransferases (DNMTs) to decrease global DNA methylation levels in CRC cells. Taken together, our results indicated that Icariside II suppressed tumorigenesis by epigenetically silencing the activation of circß-catenin-Wnt/ß-catenin axis in colorectal cancer. More importantly, the information gained from this study suggest that Icariside II may have great potential to be developed as a therapeutic drug for CRC patients.

Magnetic resonance texture analysis for the identification of cytokeratin 19-positive hepatocellular carcinoma.

PURPOSE: To investigate potential findings associated with cytokeratin 19 (CK19)-positive HCC, with special emphasis on MR texture analysis. MATERIALS AND METHODS: Forty-eight patients with CK19-negative HCC and 38 patients with CK19-positive were retrospectively evaluated by texture analysis based on conventional MRI. Clinicalpathological characteristics, conventional MR imaging findings, and the MR texture analysis contained of 2415 texture features in the seven conventional sequences were compared. Significant features for differentiating were identified by univariate and multivariate analyses. Receiver operating characteristic analyses of the significant findings were performed and compared to evaluate their diagnostic performance. RESULTS: There was no significant difference between the top1 texture feature (three-dimensional standard deviation separation of intensity on T2-weighted original images, abbreviated as: StdSeparation 3D) and the combined top1-6 feature in identifying CK19-positive HCC(P = 0.660). Univariate and multivariate analyses indicated that serum alpha-fetoprotein (AFP) level ≥400 ng/mL(P = 0.013), arterial rim enhancement(P = 0.005), and StdSeparation 3D texture character(P = 0.002) were independent variables associated with CK19-positive HCCs. The combination of the three indices showed a better performance than AFP level(P = 0.0028), arterial rim enhancement(P < 0.0001), and their combination(P = 0.0098); while no significantly better than the StdSeparation 3D texture character alone(P = 0.0788). An acceptable discrimination(AUC = 0.765) with both sensitivity and specificity greater than 75% was achieved for StdSeparation 3D texture character. CONCLUSION: Serum AFP level ≥400 ng/mL, arterial rim enhancement, and the StdSeparation 3D texture character were independently associated with CK19-positive HCC. The StdSeparation 3D texture character may be a reliable imaging biomarker which can improve the diagnostic performance.

[Quality evaluation of Magnoliae Officinalis Cortex based on combinative method of fingerprint,quantitative analysis of multicomponents and chemometrics].

This study aims to establish a combinative method based on fingerprint,assay of multi-component and chemometrics for quality evaluation of Magnoliae Officinalis Cortex. Twenty batches of samples were determined by UPLC and a common mode of fingerprint was established. The similarities between fingerprints of 20 batches of samples were over 0. 90 and the common mode were evaluated. Eight components were identified as syringing, magnocurarine, magnoflorine, magnoloside B, magnoloside A, honokiol,magnolol,and piperitylmagnolol by comparison with reference substances and their content in samples were simultaneously determined.Based on the results,the fingerprint had good consistency between the same origin and minor diversity between the different sources.Piperitylmagnolol and peak 13 could be used as a distinction with the different sources. According to content of 8 components,Fisher discriminant analysis model was established and different source sample was classified pursuant to the discriminant fraction. It is indicated that simultaneous quantification of multi components coupled with chemometrics analysis could be a well-acceptable strategy to identify and evaluate the quality of Magnoliae Officinalis Cortex.

[Effect of matrine on NO and ADMA metabolism pathways in serum and tissues of mice with lipopolysaccharide-induced intestine tissue inflammation].

OBJECTIVE: To discuss the effect of matrine on nitric oxide (NO) and asymmetric methylarginine (ADMA) metabolism pathways in serum and tissues of mice with lipopolysaccharide (LPS) -induced intestine tissue inflammation. METHOD: Kunming mice were randomly divided into five groups: the normal control group, the LPS group and matrine (80, 40, 20 mg x kg(-1) x d(-1)) groups. The mice were intragastrically administered with drugs for 3 d (distilled water of the same volume for the normal control group and the LPS group). One hour after the last intragastrical administration, normal saline or LPS (1 mg x kg(-1)) were intraperitoneally injected. Twelve hours later, serum and tissues were collected to determine NO and ADMA levels and observe the pathological changes of intestinal tissues. The Western blot method was adopted to detect the protein expressions of arginine methyltransferases 1 (PRMT1) and dimethylarginine dimethylaminohydrolase 2 (DDAH2) in intestinal tissues. RESULT: Compared with the model group, matrine (80, 40, 20 mg x kg(-1) x d(-1)) groups showed lower NO content in serum and tissues, higher ADMA level in serum and increased PRMT1 expression in intestinal tissues, but without effect on DDAH2 expression. CONCLUSION: Matrine could inhibit LPS-induced intestine tissue inflammation in mice. Its action mechanism is related to the decreased NO content in serum and tissues and increased ADMA level in serum and PRMT1 expression in intestinal tissues.

Arbuscular mycorrhizal symbiosis for sustainable cultivation of Chinese medicinal plants: a promising research direction.

Arbuscular mycorrhizal (AM) are symbiotic systems in nature and have great significance in promoting the growth and stress resistance of medicinal plants. During our literature search from the Chinese Scientific Information Database (Chinese National Knowledge Infrastructure, CNKI) we obtained 65 articles with "AM fungi" and "medicinal plant" as the key words, which indicates that in China, research efforts on these topics have been increasing. The main purposes of this review are to discuss the effects of mycorrhiza on the active ingredients of Chinese medicinal plants in comparison with results obtained in other plants in studies conducted by the international research community, and to introduce works published in Chinese journals to international colleagues.

Arbuscular mycorrhizal symbiosis and active ingredients of medicinal plants: current research status and prospectives.

Medicinal plants have been used world-wide for thousands of years and are widely recognized as having high healing but minor toxic side effects. The scarcity and increasing demand for medicinal plants and their products have promoted the development of artificial cultivation of medicinal plants. Currently, one of the prominent issues in medicinal cultivation systems is the unstable quality of the products. Arbuscular mycorrhiza (AM) affects secondary metabolism and the production of active ingredients of medicinal plants and thus influence the quality of herbal medicines. In this review, we have assembled, analyzed, and summarized the effects of AM symbioses on secondary metabolites of medicinal plants. We conclude that symbiosis of AM is conducive to favorable characteristics of medicinal plants, by improving the production and accumulation of important active ingredients of medicinal plants such as terpenes, phenols, and alkaloids, optimizing the composition of different active ingredients in medicinal plants and ultimately improving the quality of herbal materials. We are convinced that the AM symbiosis will benefit the cultivation of medicinal plants and improve the total yield and quality of herbal materials. Through this review, we hope to draw attention to the status and prospects of, and arouse more interest in, the research field of medicinal plants and mycorrhiza.

[Spectrometric determination of trace elements in anticancer new medicine Fagopyrum dibotrys].

The golden buckwheat Fagopyrum dibotrys produced in Yunnan has a unique anti-cancer effects. It is a main raw material of "Wei Mai ning" capsules which is the national second-class anti-cancer drug. The present paper used (5 : 1) mixed acid as digestive juice to process the sample, and determine the twelve elements including K, Ca, Cu, Na, Mg, Mn, Fe, Zn, Pb, Cr, Cd and Co in the Fagopyrum dibotrys by inductively coupled plasma atomic emission spectrometry(ICP-AES). The detection limits of this method were 0.017-0.084 microg x mL(-1), the RSDs (n = 8) were all 0.09%-1.87%, and the addition standard recoveries(ASR) (n = 8) were 98.2%-107.4% for all elements. The research results showed that there is rich K(1 477.3 microg x g(-1)) in the Fagopyrum dibotrys, there are not harmful elements Cd and Pb, and this result is mainly related to the geochemistry background where the sample lived. The contents of seven remaining kinds of elements ranked as Na (826.1) > Ca (765.2 > Mg (493.4) > Zn (112.7) > Fe (56.5) > Cu (11.4) > Mn (4.49 microg x g(-1)). This result provides some theoretical basis for the study of internal relations between trace elements in Fagopyrum dibotrys and efficacy. It' s also useful for better development and utilization of the resource.

Identification of zinc-finger BED domain-containing 3 (Zbed3) as a novel Axin-interacting protein that activates Wnt/beta-catenin signaling.

Axin, a key modulator of the Wnt/beta-catenin pathway, acts as a scaffold protein in phosphorylating and degrading cytoplasmic beta-catenin. Canonical Wnt proteins appear to stabilize beta-catenin by inducing the interaction of LRP5/6 with Axin. This interaction requires the phosphorylation of the Ser or Thr residues in the PPPP(S/T)PX(T/S) motifs at the intracellular domain of LRP5/6. In this work, we identified a novel Axin-interacting protein, zinc-finger BED domain-containing 3 (Zbed3), by yeast two-hybrid screening. The interaction was confirmed in co-immunoprecipitation experiment in mammalian cells and in vitro pulldown assays. Moreover, we found Zbed3 also contains a PPPPSPT motif, which is crucial to its binding to Axin. The Ser and Thr residues in the motif appear to be also phosphorylated by glycogen synthase kinase 3beta (GSK3beta) and the CKI family kinases, as GSK3beta and CKIepsilon could enhance the interaction of Zbed3 with Axin. Mutation of the Ser (SA) or Thr (TA) residue to Ala in the motif markedly impaired its ability to interact with Axin. Expressing Zbed3, but not these mutants, led to inhibition of GSK3beta-mediated beta-catenin phosphorylation, cytoplasmic beta-catenin accumulation, and activation of lymphoid enhancer binding factor-1-dependent reporter gene transcription. Furthermore, knockdown of Zbed3 with RNA interference attenuated Wnt-induced beta-catenin accumulation, lymphoid enhancer binding factor-1-dependent luciferase reporter activity, and the Wnt target gene expression. These results together indicate that Zbed3 is a novel Axin-binding protein that is involved in Wnt/beta-catenin signaling modulation.

Caprin-2 enhances canonical Wnt signaling through regulating LRP5/6 phosphorylation.

The low-density lipoprotein receptor-related proteins 5 and 6 (LRP5/6) are coreceptors for Frizzled and transmit signals from the plasma membrane to the cytosol. However, the mechanism for LRP5/6 signal transmission remains undefined. Here, we identify cytoplasmic activation/proliferation-associated protein 2 (Caprin-2) as a LRP5/6-binding protein. Our data show that Caprin-2 stabilizes cytosolic beta-catenin and enhances lymphoid enhancer-binding factor 1/T cell factor-dependent reporter gene activity as well as the expression of Wnt target genes in mammalian cells. Morpholino-mediated knockdown of Caprin-2 in zebrafish embryos inhibits Wnt/beta-catenin signaling and results in a dorsalized phenotype. Moreover, Caprin-2 facilitates LRP5/6 phosphorylation by glycogen synthase kinase 3, and thus enhances the interaction between Axin and LRP5/6. Therefore, Caprin-2 promotes activation of the canonical Wnt signaling pathway by regulating LRP5/6 phosphorylation.

Nuclear Dvl, c-Jun, beta-catenin, and TCF form a complex leading to stabilization of beta-catenin-TCF interaction.

In canonical Wnt signaling, Dishevelled (Dvl) is a critical cytoplasmic regulator that releases beta-catenin from degradation. Here, we find that Dvl and c-Jun form a complex with beta-catenin-T-cell factor 4 (TCF-4) on the promoter of Wnt target genes and regulate gene transcription. The complex forms via two interactions of nuclear Dvl with c-Jun and beta-catenin, respectively, both of which bind to TCF. Disrupting the interaction of Dvl with either c-Jun or beta-catenin suppresses canonical Wnt signaling-stimulated transcription, and the reduction of Dvl diminished beta-catenin-TCF-4 association on Wnt target gene promoters in vivo. Expression of a TCF-Dvl fusion protein largely rescued the c-Jun knockdown Wnt signaling deficiency in mammalian cells and zebrafish. Thus, we confirm that c-Jun functions in canonical Wnt signaling and show that c-Jun functions as a scaffold in the beta-catenin-TCFs transcription complex bridging Dvl to TCF. Our results reveal a mechanism by which nuclear Dvl cooperates with c-Jun to regulate gene transcription stimulated by the canonical Wnt signaling pathway.

[Effects of N P K on yield and baicalin contents of Scutellaria baicalensis].

OBJECTIVE: To find out the effects of different levels and combinations of nitrogen, phosphorus and potassium on the growth and secondary metabolites of Scutellaria baicalensis. METHODS: The quantities of growth, physiological indicators and baicalin contents in different fertilization treatment were calculated and compared. RESULTS: The growth physiological indicators and baicalin contents of S. baicalensis in complex fertilization treatment were better than those in single element fertilization treatment, and nutrition balance types better than maladjustment type. The reasonebly fertilitzing in complex fertilization treatment, nitrogen could improve the fertilizer efficiency of phosphorus. CONCLUSION: According to the fertilization mathematic model, the optimum fertilization dosage should be primarily defined as N: 22.6-25.0 g/m2, P: 30.6-44.6 g/m2, K: 5.4-21.4 g/m2, that is 2N:3P:1K.

[Present situation and prospects of special fertilizer for traditional Chinese medicine herbs].

OBJECTIVE: To find out the present situation and the development trend special fertilizer of the traditional Chinese medicina plants. METHOD: By consulting a great deal of literatures on special fertilizer and fertilization on traditional Chinese medicine herbs, and based on the scientific research and manufacture experience of the author, and the theoretic actuality of the researches on the fertilization of traditional Chinese medicine herbs, the present study of the special fertilizer inside and outside of our country was analyzed. CONCLUSION: The view points of developing special fertilizer for Chinese traditional medicine were put forward, and the development trend of special fertilizer for traditional Chinese medicine herbs was forecasted.

[The investigation of the licorice resources in northeast China].

OBJECTIVE: To find out the licorice(Glycyrrhiza uralensis) distribution area, resource complexion and resource reserves of northeast China, to analyze the cause of the swing of the pendulum of resources, to put forward the countermeasure of resource protection and to provide evidence for the establishment of relative statutes. METHOD: Combination of visit-inquisition and sample-square investigation involving the resource complexion of the 32 counties and cities in the northeast China wasmade. RESULT AND CONCLUSION: The east distribution boundary and the whole distribution current of licorice in the northeast China were determined the northeast licorice distributing region was compartmentalized into three typical sub-regions, and the licorice population character and artificial disturbing status in main counties of every sub-region were described. The licorice reserves were also figured out. At the same time, the nature and the artificial factors that influenced the swing of the pendulum of licorice resource were analyzed, and the correlative safeguard measure was brought forward.

Effects of Deleting the Amino-terminal Domain of GRK-2 on Its Function.

To reveal the possible role of the amino-terminal domain of G protein-coupled receptor kinases(GRKs)in receptor phosphorylation and/or modulation of its kinase activity, a truncated mutant of GRK-2 lacking the amino-terminal domain(deltaN-GRK2)was made. deltaN-GRK2 was expressed effectively in E.coli as a GST fusion protein and was purified by affinity chromatography on a GSH-Sepharose column. deltaN-GRK2 was then separated from GST tag by thrombin cleavage and recovered. Although deltaN-GRK2 had nearly identical activity with wild-type GRK-2 in phosphorylation of peptide substrate, it completely lost the ability to phosphorylate the light-activated receptor rhodopsin. Furthermore, deletion of the amino-terminal domain rendered GRK-2 unresponsive to the regulation of kinase activity by a truncated form of rhodopsin, (329)G-Rho(*) and beta gamma subunits of G protein. These results demonstrated that the amino-terminal domain was necessary to GRK2 for both the phosphorylation of receptor and the regulation of its kinase activity by the receptor. It was reasonable to postulate that this domain has little, if any effect on the catalytic domain of natural form of GRK2.