OBJECTIVETo determine the interaction between miR-21 and DNA methylation in different breast cancer cells.

METHODSFluorescence tagged miR-21 inhibitor and its negative control (NC) were transient transfected into MCF-7 and MDA-MB-231 cell, the transfection efficiency was observed using fluorescence microscopy, and the miR-21 expression level and genome DNA methylation status before and after transfection were assessed by real-time PCR and bisulfite-qMSP respectively. To investigate the regulation effect of DNA methylation on miR-21, cells were treated with 5-AZA (2.5 µmol/L) for 72 h, with dimethyl sulfoxide (DMSO) treatment as its negative control (NC), and the expression level of phosphatase and tensin homolog deleted on chromosome ten (PTEN) and AKT(also known as Protein Kinase B), two downstream genes of miR-21 were detected by Western blot.

RESULTSThe expression of miR-21 in MCF-7 cell was significantly knocked down (P < 0.01) by miR-21 inhibitor, with the genome DNA methylation level (P < 0.05) and all the three Dnmts: Dnmt1, Dnmt3a, and Dnmt3b unregulated. In contrast, the miR-21 expression in MDA-MB-231 cell was elevated ( P < 0.01) by miR-21 inhibitor, meanwhile, down- regulated of genome DNA methylation (P < 0.05) and Dnmt3b expression, upregulation of Dnmt3a were also observed. In addition, treated with 5-AZA resulted in significant increases of miR-21 expression in both MCF-7 and MDA-MB-231 cells (P < 0.01), with the protein level of PTEN increased in MCF-7 cell, which was further involved in the downregulation of AKT.

CONCLUSIONThe regulation effects of DNA methylation by transient transfection of miR-21 in MCF-7 and MDA-MB-231 cells are almost opposite, whilst the expression of miR-21 in two cell lines were all upregulated by decreased DNA methylation level and our results may provide some experimental evidences for the future development of rational therapy for different breast cancer.

Azacitidine ; Breast Neoplasms ; genetics ; Cell Line, Tumor ; DNA (Cytosine-5-)-Methyltransferase 1 ; DNA (Cytosine-5-)-Methyltransferases ; metabolism ; DNA Methylation ; Down-Regulation ; Gene Expression Regulation, Neoplastic ; Humans ; MCF-7 Cells ; MicroRNAs ; genetics ; PTEN Phosphohydrolase ; metabolism ; Promoter Regions, Genetic ; Proto-Oncogene Proteins c-akt ; metabolism ; Real-Time Polymerase Chain Reaction ; Up-Regulation

The phosphorylation is one of most common protein post-translational modifications. The protein phosphorylation plays important roles in the life through the reversible process of phosphorylation and dephosphorylation by kinases and phosphatases. Systematical analysis of the phosphorylation state of proteins would greatly help to reveal the mystery of the life. Recently, with the development of mass spectrometer, bioinformatics sortwares and enrichment methods of phosphopeptides, phosphorylation stduy of orgnism proteins by mass spectrometer has become mature gradually. Liver is one of the most important metabolic and immune organs. In-depth study of protein phosphorylation in liver is of great importance to reveal its function. And booming phosphoproteomics has been applied into the study of liver, which has deepened the knowledge of molecular mechnism of its physiology and pathology states. Here, we review the recent progress on the research and development of phosphoproteomics and their application in liver proteomics study.
Computational Biology ; Humans ; Liver ; metabolism ; pathology ; physiology ; Mass Spectrometry ; Phosphopeptides ; metabolism ; Phosphorylation ; Protein Processing, Post-Translational ; Proteomics

This study deals with colony expansion characteristics, biomass increment and Na accumulation in mycelia of three ectomycorrhizal fungi ( Boletus edulis, Xerocomus chrysenteron and Gomphidius viscidus ) under treatment of Na2SO4 and NaCl. The experimental results showed that no impact was observed from Na2SO4 and NaCl treatments on the growth mode of the mycelia, but the biomass increment of X. chrysenteron and G. viscidus was significantly restrained under 0.1 mol/L Nacl. Under the treatment of Na2SO4, biomass increment of X. chrysenteron is significantly higher than the control, but the biomass of G. viscidus is lower than that of the control, and no significant impact was measured on growth of B. edulis under both treatments of Na2 SO4 and NaCl. The experiment also indicated that Na accumulation in mycelia varied significantly among the three tested strains, highest Na accumulation was measured in X. chrysenteron under treatment of NaCl, while under the treatment of Na2SO4 , accumulation of Na in B. edulis is much higher than the other two strains.

Objective To prospectively evaluate the value of US,MSCT,EUS and MRI in the quantitative evaluation of the extent of pancreaticobiliary duct obstruction in pancreatic cancer.Methods Consecutive 68 patients with pancreatic carcinoma underwent US,MSCT,EUS and MRI before surgery.The diameter of extrahepatic bile duct and pancreatic duet were measured,and correlation analysis was performed with surgical specimens.Results Diameters of extrahepatic bile duct scaled by US.MSCT,EUS and MRI were(16.60±6.33)mm,(18.90±6.74)mm,(18.80±5.88)nun and(17.26±4.83)mm,and diameter measured from surgical specimens was(18.39±6.05)mm;the correlation among the four imaging examinations and the surgical evaluation were r=0.3839,P=0.1055;r=0.7113,P=0.0011; r=0.3759,P=0.0465;r=0.3376,P=0.2872,respectively. Kappa Values were 0.6285,0.7115,0.6661 and 0.7490,respectively.The diameter of pancreatic duct was(15.90±3.41)mm,(6.83 4-3.70)mm,(6.77±3.22)mm and(5.58±2.65)mm,and diameter measured from surgical specimens was(5.97±2.60)mm,the correlation among the four imaging examinations and the surgical evaluation were r=0.3584,P=0.2895;r=0.6148,P＜0.0001; r=0.7373,P＜0.0001;r=1.0746,P＜0.0001.Kappa values were 4.159,9.094,9.001 and 4.050.All of these parameters were in coherence with surgical findings.Condusions US could be used as the initial method in the assessment of extrahepatic and pancreatic duct obstruction.MRI and MSCT,combined with EUS if necessary,could be used to quantitatively evaluate the extent of pancreaticobiliary obstruction.

Objective To explore clinieopathologieal features and prognosis in a high grade malignancy group of colorectal mueinous carcinoma, signet-ring cell carcinoma and low-differentiated carcinoma. Methods Retrospective analysis and follow-up were made on 148 patients with colorectal mucinous carcinoma, 55 patients with signet-ring cell carcinoma and 281 low- differentiated carcinoma treated in our hospital from 1988 to 2006. Obtained data were analyzed by SPSS13.0. Related factors underwent x2 analysis, survival analysis were estimated using Kaplan-Meier method and compared using the Log-rank test. Results Coloreetal signet-ring cell carcinoma were significantly different from mucinous carcinoma and low- differentiated carcinoma in gender, age, tumor location, bowel obstruction, operative modus, tumor embolism, tumor infiltration and lymph node metastasis(x2 = 7, 67,38.4,86.0,14.5,93.7,17.3,62. 1,24. 4,56.17 ,P <0.05). Median survival time was 24 months in signet-ring cell carcinoma, 47 months in mueinous carcinoma and 49 months in low- differentiated carcinoma. The difference is of significance(x2 = 21.3, P < 0.05). Conclusions Clinicopathological characteristics and prognosis of colorectal signet-ring cell carcinoma, mueinous carcinoma and low- differentiated carcinoma is of significant difference(P < 0.05). Malignancy of signet-ring cell carcinoma is higher with worse prognosis.

Objective To explore the clinical diagnosis and management of hepatecellular adenoma(HA). Methods Seventeen HA cases treated in our hospital from Janurary 1986 to June 2007 were studied retrospectively.The clinical and pathologic data were reviewed. Results Of 17 patients,9 were male and 8 were female with median age of 46 years.None of the female patients had a history of oral contraceptives.Most patients were agymptomatic.Preoperative liver function test and AFP serum test were normal.Preoperative imaging study yielded deftnite diagnosis in only a fraction of patients.All 17 cases underwent surgical resection and the postoperative course is uneventful.All cases were followed up ranging from 6 to 252 months.Recurrence was not found.Pathology reported atypical hyperplagia in 2 cases and with malignant transformation in 1 case. Conclusions The diagnosis of HA is exclusive and dependent on clinical data,laboratory test and radiography.HA has a tendency to malignancy.Surgery is recommended in all cases.

Chemical constituents of ethyl acetate extract of Sapium sebiferum leaves were isolated and purified by various chromatographic methods, including column chromatographies over silica gel, macroporous adsorption resin, and Sephadex LH-20, as well as preparative TLC and semi preparative HPLC. As a results, 15 compounds were separated from Sapium sebiferum leaves and their structures were examined by spectral analysis including NMR and MS data and identified as( + )-(7R,7'R,7"S,7'"S,8S,8'S,8"S,8'"S)-4", 4"'-dihydroxy-3,3',3",3',5,5'-hexamethoxy-7,9';7',9-diepoxy-4,8";4',8'"-bisoxy-8,8'-dineo-lignan-7",7"',9",9"'-tetraol(1) ,1-(4'- hydroxy-3'-methoxyphenyl)-2-[4"-(3-hydroxypropyl) -2", 6"-dimethoxyphenoxy] propane-1, 3-diol (2), Thero-2, 3-bis-(4-hydroxy-3- methoxypheyl)-3-methoxy-propanol(3) , threo-5-hydroxy-3,7-dimethoxyphenyl propane-8,9-diol (4), boropinol B (5), threo-8S-7-methoxysyringylglycerol(6), 5-hydroxymethylfurfural(7), 5-( methoxy-methyl)-1H-pyrrole-2-carbaldehyde (8), quercetin (9) , kaempferol (10), ethyl gallate(11), coniferaldehyde(12), vanillin(13), 7-hydroxy-6-methoxy-2H-1-henzopyran-2-one(14),and 1-heptacosanol (15). All compounds except for compounds 9-11,14 were separated from this plant for the first time.
Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Mass Spectrometry ; Molecular Structure ; Plant Leaves ; chemistry ; Sapium ; chemistry

0.05),the mean FA on the left was higher than the right(t=1.912,P

As one of the most serious types of psoriasis, pathogenesis of erythrodermic psoriasis (EP) is unclear so far. In this study, we aimed to detect the levels of Th1/Th2 cytokine-associated transcription factors and T-lymphocyte clone in peripheral blood mononuclear cells (PBMCs) derived from EP patients, and gene expression level of T-bet/GATA-3 in skin lesion. The potential role of Th1/Th2 reaction pattern played in the pathogenesis of EP was also discussed. Serum levels of IFN-γ, IL-2, IL-4 and IL-10 were quantified by ELISA among 16 EP patients, 20 psoriasis vulgaris (PV) patients and 15 healthy controls. The expression levels of T-bet/GATA-3 in the skin lesion and PBMCs were examined by real-time qPCR. The ratio of Th1/Th2 was measured by flow cytometry. The levels of IFN-γ, IL-2, IL-4 and IL-10 were higher in EP patients than in the healthy controls. The levels of IL-4 and IL-10 were 69.44±11.45 and 12.62±4.57 pg/mL, respectively, in EP patients, significantly higher than those in PV patients and healthy controls (P<0.05). Flow cytometry revealed the levels of both Th1 and Th2 in PBMCs from EP patients were higher than those in healthy controls, and the Th1/Th2 ratio was dramatically lower than in PV patients (P<0.01). The ratios of IFN-γ/IL-4 and T-bet/GATA-3 in EP patients were both less than 1.0, suggesting a reversal when compared with the other two groups. Our study indicated that the EP patients exerted a Th1/Th2 bidirectional response pattern, and the balance of Th cell subsets inclines to Th2, which might be one of the important mechanisms of EP pathogenesis.

Polymeric micelles which are self-assembled from amphiphilic copolymers are thermodynamically stable, and they can solubilize hydrophobic drugs by the hydrophilic core. Many excellent active compounds are confined because of general low oral bioavailability due to poor solubility. Take into account from the two points above, polymeric micelles may be used as proper oral carrier to improve the dissolubility of hydrophobic drugs, and enhance the permeation though gastrointestinal tract, therefore, the pharmacodynamics is elevated. Meanwhile, the segments in copolymers are multivariate, so many kinds of micelles can be obtained, such as, pH- or thermo- sensitive as well as mucoadhesive ones. The modified micelles can alter drug release profiles while solubilizing them, that is why the oral bioavailability increase further. In this review, recent progress of polymeric micelles used in oral administration is summarized, and the prospect of polymeric micelles' application in this field is also evaluated.
Administration, Oral ; Animals ; Area Under Curve ; Biological Availability ; Drug Carriers ; Drug Delivery Systems ; Humans ; Micelles ; Pharmaceutical Preparations ; administration & dosage ; Poloxamer ; chemistry ; Polyethylene Glycols ; chemistry ; Polymers ; chemistry ; Risperidone ; administration & dosage ; pharmacokinetics ; Silymarin ; administration & dosage ; pharmacokinetics ; Solubility