Objective To study if 5-Aza-2’-deoxycytidine along or together with 4-phenylbutyric acid could affect miR-196b expression levels in chronic myeloid leukemia cells .Methods K562 cells were treated with DNA methylation inhibitor 5-Aza-2’-deoxycytidine, histone deacetylase inhibitors 4-phenylbutyric acid separately and the combined treatment with both of them, then expression levels of miR-196b were detected using Real-time PCR.Results The half inhibition concentration of 4-phenylbutyric acid was 1.58mmol/L.Comparing with the expression level of miR-196b in normal human bone marrow cells, the expression levels of miR-196b were significantly lower in Aza group , PBA group and negative control cells and nearly consistent among three groups , and as high as normal cells in combined treatment group . Conclusion The expression level of miR-196b in K562 cells could not return to normal treated with 5-Aza-2 ’-deoxycytidine or 4-phenylbutyric acid separately , while could restore normal when treated with both agents , indicating that miR-196b expression level in K562 cells is related with both DNA methylation and histone acetylation .

Objective To study the application of Cell Counting Kit-8(CCK-8) in detecting the growth inhibiting effect of 5-Aza-2’-deoxycytidine on chronic myeloid leukemia cell .Methods The proliferation of K562 cells was detected by CCK-8 with different concentrations of 5-Aza-2 ’-deoxycytidine and the cell cycle and apoptosis of K 562 cells were detected after K562 treated by 50% inhibitory concentration of 5-Aza-2 ’-deoxycytidine .Results The 50% inhibitory concentration of 5-Aza-2’-deoxycytidine was 15.55nmol/L, after treated with this concentration , K562 cells showed that G2 phase arrest occurred , proliferation inhibited and apoptosis peaks appeared .Conclusion Inhibition of proliferation of K562 cells with different concentrations of 5-Aza-2’-deoxycytidine varied in a dose-dependent relationship , and 5-Aza-2’-deoxycytidine could promote apoptosis of K 562 cells.CCK-8 can be used in detecting the effect of 5-Aza-2’-deoxycytidine on chronic myeloid leukemia cells .

Object: To study the proliferation of hCTGF on cells and its biological function on bone injury healing.Methods: The fibroblast with potential differentiation was transfected by eukaryotic gene delivery system and then transferred into the experimental animal model with bone fracture.The data were collected by molecular biological and clinical orthopedic technique detection analysis.Results: The results demonstrated an obvious proliferation of hCTGF on cells,suggesting that hCTGF have the biological activity of repairing bone injury via gene therapy.The results provide a new activity factor and treatment approach for bone injury in clinics.

Four kinds of ionic liquids [BMIM] Br, [BMIM] BF4, [BMIM] PF6, [HMIM] PF6 were used to analyze the content of oleanic acid and paeoniflorin in Paeonia lactiflora with ultrasonic-assisted extraction coupled with HPLC. The chromatographic column, Purospher star RP-C18 (4.6 mm x 250 mm, 5 μm), was used. Acetonitrile and water (90:10) as mobile phase was used to determine the content of oleanic acid with a gradient elution and flow rate at 1.00 mL · min(-1), detection wavelength at 210 nm, chromatographic column temperature at room temperature. Paeoniflorin content was determined using acetonitrile and water (18:82) as mobile phase with a gradient elution and flow rate at 1.00 mL · min(-1), detection wavelength at 250 nm, the chromatographic column temperature at room temperature. The result show that oleanic acid has the highest extraction yield when the conditions are solid-liquid ratio of 1:80 (g · mL(-1)), and the [BMIM] Br methanol solution concentration of 0.6 mol · L(-1). Under the optimal extraction conditions, the content of oleanic acid from 0.24 to 3.76 μg showed a good linearity (r = 0.9999), the average recovery was 97.20%. Paeoniflorin has the highest extraction yield when the conditions are solid-liquid ratio of 1:130 (g · mL(-1)), and the [C4 MIM] PF6 methanol solution concentration of 0.6 mol · L(-1). Under the optimal extraction conditions, paeoniflorin content from 0.42 to 4.20 μg showed a good lin- earity (r = 1.000), the average recovery was 98.84%. This method is simple and reliable, its repeatability is also very good. It has important significance in the study P. lactiflora of ionic liquid microextraction.
Chromatography, Reverse-Phase ; methods ; Glucosides ; analysis ; Ionic Liquids ; chemistry ; Monoterpenes ; analysis ; Oleanolic Acid ; analysis ; Paeonia ; chemistry ; Ultrasonics

OBJECTIVETo study the chemical constituents of paeoniflorins from root cortex of the Paeonia suffruticosa.

METHODPaeoniflorins were isolated by partition, silica gel column chromatography, medium-pressure RP chromatography and HPLC. Their structures were elucidated by spectroscopic methods including IR, MS and NMR techniques.

RESULTNine paeoniflorins were isolated and identified from the root bark ethanol extract of P. suffruticosa.

CONCLUSIONThe new compound was identified as paeoniflorin-4-ethyl ether, which may be an artifact produced during extraction proved by simulate experiment.

Benzoates ; chemistry ; isolation & purification ; Bridged-Ring Compounds ; chemistry ; isolation & purification ; Glucosides ; chemistry ; isolation & purification ; Molecular Structure ; Paeonia ; chemistry ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry

OBJECTIVETo study the chemical constituents in rhizome of Pinellia ternata.

METHODThe constituents were isolated by silica-gel and Sephadex LH-20 chromatography. The structures were identified by spectroscopic analysis including 2D NMR techniques.

RESULTSix compounds were obtained and identified as stigmast-4-en-3-one(I), cycloartenol(II), 5alpha,8alpha-epidioxyergosta-6,22-dien-3-ol(III), beta-sitosterol-3-O-beta-D-glucoside-6'-eicosanate(IV), alpha-monpalmitin(V), beta-sitosterol(VI). The bioactive assay indicated that: compound III was active against the human tumor cell lines HCT-8, Bel-7402, BGC-823, A549, A2780.

CONCLUSIONCompounds I-IV were isolated from Pinellia ternata for the first time, compound II was the first triterpene isolated from this genus. Compound III may be one of the antitumor constituents of P. ternata.

Antineoplastic Agents, Phytogenic ; chemistry ; isolation & purification ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Ergosterol ; analogs & derivatives ; chemistry ; isolation & purification ; pharmacology ; Humans ; Phytosterols ; chemistry ; isolation & purification ; Pinellia ; chemistry ; Plants, Medicinal ; chemistry ; Rhizome ; chemistry ; Stigmasterol ; analogs & derivatives ; chemistry ; isolation & purification ; Triterpenes

Liver regeneration depends on powerful immune system of ownself. TNF-α,IFN-γ,IL-6,IL-12,etc. that secreted by innate immune cells such as macrophages,dendritic cells,NK cells and NKT cells,could act on the hepatocytes and regulate liver regen-eration (LR) through corresponding signaling pathways. This article summarizes the mechanism of different innate immune cells on hep-atocytes,clarifies the recent advances of liver innate immune cellsduring liver regeneration process,lay the foundation for revealing the molecular mechanism of the development of liver regeneration and liver diseases, and for the research and development of new therapeutic methods for liver diseases.

Objective of this study was to perform bioinformatics analysis of the characteristics of gene expression profiling regulated by amifostine and predict its novel potential biological function to provide a direction for further exploring pharmacological actions of amifostine and study methods. Amifostine was used as a key word to search internet-based free gene expression database including GEO, affymetrix gene chip database, GenBank, SAGE, GeneCard, InterPro, ProtoNet, UniProt and BLOCKS and the sifted amifostine-regulated gene expression profiling data was subjected to validity testing, gene expression difference analysis and functional clustering and gene annotation. The results showed that only one data of gene expression profiling regulated by amifostine was sifted from GEO database (accession: GSE3212). Through validity testing and gene expression difference analysis, significant difference (p < 0.01) was only found in 2.14% of the whole genome (460/192000). Gene annotation analysis showed that 139 out of 460 genes were known genes, in which 77 genes were up-regulated and 62 genes were down-regulated. 13 out of 139 genes were newly expressed following amifostine treatment of K562 cells, however expression of 5 genes was completely inhibited. Functional clustering displayed that 139 genes were divided into 11 categories and their biological function was involved in hematopoietic and immunologic regulation, apoptosis and cell cycle. It is concluded that bioinformatics method can be applied to analysis of gene expression profiling regulated by amifostine. Amifostine has a regulatory effect on human gene expression profiling and this action is mainly presented in biological processes including hematopoiesis, immunologic regulation, apoptosis and cell cycle and so on. The effect of amifostine on human gene expression need to be further testified in experimental condition.
Amifostine ; pharmacology ; Computational Biology ; Gene Expression ; drug effects ; Gene Expression Profiling ; methods ; Humans ; Microarray Analysis ; Molecular Sequence Annotation

OBJECTIVETo investigate the chemical constituents of the red alga Gymnogongrus flabelliformis Harv.

METHODCompounds were isolated by normal phase silica gel and Sephadex LH - 20 gel column chromatography and reverse phase HPLC. Their structures were elucidated by spectroscopic methods including MS, 1H-NMR, 13C-NMR. Cytotoxicity of the compounds was screened by using standard MIT method.

RESULTFive compounds were isolated from G. flabelliforrmis, their structures were identified as(3S, 6R, 7E)-( + )-3-hydroxyl-4, 7-mega-stigmadien-9-one (1), (3S, 5R, 6S, 7E)-(-)-3-hydroxy-5, 6-epoxy-7-megastigmene-9-one (2), (3S, 5S, 6R, 7E)-(+)3-hydroxy-5, 6-epoxy-7-megastigmene-9-one (3), dehydrovomifoliol (4), (3R)-(-)4-[(2R, 4S)-4-acetoxy-2-hydroxy-2, 6, 6-trimethylcyclohexylidene] -3-buten-2-one (5).

CONCLUSIONAll of the compounds were obtained from this species for the first time and compound 1 was a new natural product. These compounds were inactive (IC50 > 10 microg x mL(-1)) in the MTT assay against several human cancer cell lines.

Antineoplastic Agents ; chemistry ; isolation & purification ; pharmacology ; Butanols ; chemistry ; isolation & purification ; pharmacology ; Cell Line, Tumor ; Cell Survival ; drug effects ; Chromatography, High Pressure Liquid ; Cyclohexanones ; chemistry ; isolation & purification ; pharmacology ; Humans ; Inhibitory Concentration 50 ; Magnetic Resonance Spectroscopy ; Norisoprenoids ; chemistry ; isolation & purification ; pharmacology ; Rhodophyta ; chemistry

OBJECTIVETo investigate the chemical constituents of red alga Corallina pilulifera.

METHODCompounds were isolated by normal phase silica gel and Sephadex LH - 20 gel column chromatography, reverse phase HPLC and recrystallization. Their structures were elucidated by spectroscopic methods including MS, 1H-NMR, 13C-NMR, HSQC, HMBC. Cytotoxicity of the compounds was screened by using standard MTT method.

RESULTSeven compounds were isolated from red alga C. pilulifera, their structures were identified as (E) -phytol epoxide (1), phytenal (2), phytol (3), dehydrovomifoliol (4), loliolide (5), 3beta-hydroxy-5alpha, 6alpha-epoxy-7-megastigmene-9-one (6), 4-hydroxybenzaldehyde (7).

CONCLUSIONAll of the compounds were obtained from this species for the first time. These compounds were inactive (IC50 > 10 microg x mL(-1)) in the MTT assay.

Benzaldehydes ; chemistry ; isolation & purification ; pharmacology ; Benzofurans ; chemistry ; isolation & purification ; pharmacology ; Cell Line, Tumor ; drug effects ; Humans ; Phytol ; chemistry ; isolation & purification ; pharmacology ; Rhodophyta ; chemistry