Purpose:To study the expression and clinical significance of nm23 protein in human breast carcinoma.Methods:The expression of nm23 were detected by immunohistochemical S P methods in tissue samples from 65 cases of breast carcinoma with full follow up .Their prognostic value was analyzed by Cox regression model.Results:Positive expression of nm23 was 55.4% in the 65 breast carcinoma specimens. In the total number of patients, the 5 year overall survival rate(OS) and disease free survival rate(DFS) were 63.1% and 52.3%,respectively. Expression of nm23 was negatively associated with distant metastasis( P =0.0003) and positively with DFS and OS( P =0.0034, P =0.0001). In both monovariate Kaplan Meier plot and multivariate Cox regression analysis, the axillary lymph node status and expression of nm23 were two important prognostic factors of breast carcinoma. The hazard ratios of axillary lymph node status and expression of nm23 were 3.1046 and 0.3704, respectively. Conclusions:In breast carcinoma, secondary to the axillary lymph node status, the expression of nm23 could be an independent prognosticator, which would be helpful in selecting the high risk cases for further treatment.

An automated analytical method for simultaneous determination of vitamin A and E in livers, fortified infant formulae and eggs has been developed based on on-line solid phase extraction (SPE) coupled with a dual gradient high performance liquid chromatography system with column-switching. Firstly, food samples were centrifuged after saponified in mixture solution of anhydrous alcohol, potassium hydroxide and ascorbic acid at 80 ℃ for 30 min. Secondly, the saponified sample was loaded and washed on the first dimension extraction column using methanol-water (60∶40, V/V). Afterwards, the targeted analytes were trapped and enriched on the SPE column. Finally, the trapped analytes were transferred to the second dimension analysis column by valve-switching technique for the following separation and determination. Several key factors such as the type of SPE columns, elution buffer as well as pH of washing solution were optimized. The results showed that the calibration curves of vitamin A and E were linear in the range of 0 . 02-20 mg/L with correlation coefficient (R2) more than 0. 9998. In addition, the limits of detection (S/N=3) were found in the range of 3. 0-30. 0 μg/L. The spiked recoveries of the vitamin A and E from livers, eggs and fortified infant formulae ranged from 87 . 3% to 115 . 0% with the relative standard deviations ( RSDs ) of 1 . 8% -4 . 6%. The developed method is simple, sensitive and rapid to determine vitamins A and E in animal derived food.

AIM: To establish HPLC-fingerprints and quantitatively determine platycodin-D from Platycodon grandiflorum.METHODS: HPLC analysis was carried out on Hypersil C18 column(250 mm ? 4.6 mm,5 ?m),with a mobile phase of acetonitrile-0.05 mol/L phosphoric acid system,gradient elution,with a flow at 0.5 mL/ min,an ultraviolet detection wavelength was at 210 nm for fingerprint and at 206 nm for platycodin-D,column tem-perature at 30 ℃.RESULTS: Twelve common peaks were identified in chromatograms with reference to platycod-in-D peak from the 18 batches of the samples.CONCLUSION: The method of the HPLC-fingerprint and quantita-tive analysis is rapid,simple and accurate with a good reproducibility and can be used for the quality control of Platycodon grandiflorum.

Objective To evaluate the clinical significance of telomerase in the diagnosis of lung cancerusing SROC curve method.Methods Looking“telomerase”and“lung cancer”as keywords,retrieving journalspublished within past 20 years in order to incorporate into literatures and to collect data .To conduct the SROC analysisusing Meta -DiSc 1.4 software.Results 1.Twenty -two documents were sampled as tissue specimensand the heterogeneity was relatively large (P =0.017).To analyze the data with random effective models ,thecombined sensitivity and specificity were 0.788(0.761 -0.814)and 0.955(0.936 -0.969),respectively.TheSROC AUC area under the curve was 0.9515,SE(AUC) =0.0145.2.There was no heterogeneity(P =0.633)amongthe 10 lavaged literatures.By use of fixed effects model for data analysis ,the combined sensitivity and specificitywere 0.777(0.734 -0.816) and 0.922(0.888 -0.948),and SROC AUC area under the curve was0.9369,SE(AUC) =0.0141.Conclusion Telomerase is a ideal tumor marker,and the detection of telomeraseactivity in lavage fluid is stable and accurate in clinical diagnosis .

Objective To observe the expression of GATA-3 and IL-13 in mice with radiationinduced lung fibrosis,and to study the function of GATA-3.Methods A total of 63 C57BL/6 female mice were randomly into 2 groups,including 21 mice in control group and 42 mice in irradiated group.The thoraces of mice in irradiated group were exposed with 12 Gy of X-rays.All the mice were sacrificed at 1 h,and 1,2,4,8,16 and 24 weeks post-irradiation.The lung issues were stained by using HE and Masson methods to determine the histological changes.The expression of IL-13 in serum,and the expression of hydroxyproline and the mRNA and protein of GATA-3 in lung tissue were assayed.Results Compared with control group,there was a significant histological and pathologic change in irradiated group.The content of hydroxyproline in irradiated group was significantly higher than that in control group( Z =3.14,P ＜0.05).The expressions of GATA-3 and IL-13 were found in mice post-irradiation.Without causing conspicuous fibrotic pathological changes,there was a significantly elevated expression of Th2-specific transcription factor GATA-3 mRNA at 1 and 2 weeks post-irradiation ( t =6.50,6.33,P ＜ 0.01 ),while the expression of IL-13 reached the maximal value in serum at 16 weeks post-irradiation( t =32.21,P ＜0.01 ).Conclusions GATA-3 might play a role in promoting radiation-induced pulmonary fibrosis by upregulation of expression of Th2 cytokine IL-13.

Objective To investigate the antioxidation activity and resistance of lipid peroxidation of panax notoginseng flower total saponins.Methods The panax notoginseng flower buds were extracted with ethanol. The hydroxyl free radical(?OH), 1,1-diphenyl-2-picrythydrazyl radical(DPPH)clearing rate and resistance of lipid peroxidation of rat liver induced by Fe2+-cysteine were determined by spectrophotometry. Results Half clearance of hydroxyl free radical and DPPH. by panax notoginseng flower total saponin was 0.035 mg/ml and 0.094 mg/ml, the maximum inhibition rate of lipid peroxidation of rat liver induced by Fe2+-cysteine was 89.31%, therefore moderate concentration of extracts had a strong inhibitory effect on lipid peroxidation. Conclusions Panax notoginseng flower total saponins have antioxidant activity and resistance of lipid peroxidation.

A method for determination of residues of 26 β2-agonists in pork liver was developed using high performance liquid chromatography with tandem mass spectrometric ( HPLC-MS/MS ) . After enzymatic hydrolysis with β-Glucuronidase/Arylsulfatase for 12 hours, the pH of sample solution was adjusted to 1 using perchloric acid for protein precipitation. The precipitate was extracted with 0. 1mol/L perchloric acid aqueous. The extracts in the above two steps were combined and adjusted to pH 4 for the solid phase extraction ( MCX) . And then the 26 β2-agonists residues in the extracts were separated on a reversed phase HPLC column using a gradient elution program of 0. 1% formic acid aqueous solution (A) and 0. 1% formic acid in acetonitrile solution ( B) . Multiple reaction monitoring ( MRM) with positive polarity was selected to monitor qualitative and quantitative ion. Based on the optimized method, 26 β2-agonists could be analyzed in 15 min. The recoveries ranged from 64 . 0% to 112 . 7% for the 26 kinds ofβ2-agonists residues with three spiked levels of 5, 10 and 20 μg/kg. The relative standard deviations ( RSDs) were less than 15. 2%. The limits of detection (LOD) for the 26 kinds of β2-agonists were 0. 15-1. 35 μg/kg.

Objective To establish the mitochondrial DNA depleted cell line (ρ0 cells) of lung cancer cell A549 and to observe the radiosensitivity difference between ρ0 cells and normal A549 cells (ρ + cells).Methods The ρ0 cells were depleted of mitochondrial DNA by culturing chronically in the presence of low concentration of ethidium bromide (EB),and then the cell model was confined.Radiosensitivity of both ρ0 cells and ρ + cells was detected using the cologenic formation assay.After 6 MV X-ray irradiation in vitro,cell cycle distribution and reactive oxygen species (ROS) level were analyzed by flow cytometry and fluorescence microplate reader,respectively.Results A ρ0 cell line was successfully established and had a lower radiosensitivity than ρ + cells (t =12.57,P ＜ 0.01).After irradiation with a dose of 8 Gy,compared to ρ+ cells,ρ0 cells showed prolonged G2 arrest with less cells in G2 (t =6.82,P ＜ 0.01) and had lower increase of ROS level (t =14.51,P ＜ 0.01).Conclusions ρ0 cells have a lower radiosensitivity than ρ + cells,in which the reduction of ROS production and the prolongation of G2 arrest post-irradiation may be involved.

Objective To investigate the relationship between single nucleotide polymorphism in estrogen metabolizing genes CYP17、CYP19 and breast cancer susceptibility.Methods A case-control study was performed.PCR-base restriction fragment length polymorphism(PCR-RFLP)and short tandem repeat polymorphism(STRP)assay were used to detect the single nucleotide polymorphism of CYP17、CYP19 in 213 breast cancer cases and 430 matched controls.Resuits CYP17 A2/A2 genotype was found in 6.7%of breast cancer cases,which was significantly higher(P＜0.05)than that in controls(2.4%);the frequency of A2 allele of CYP17 was 16.2%in breast cancer cases,which was significantly higher(P＜0.05)than that in controls(10.6%).There Was alSO significant difference in the frequency of(TTTA)10allele of CYP19 which was 12.4%in breast cancer cases and 8.2%in controls(P=0.02).Conclusions The allele of CYP17 A2 and CYP19(TTTA)10 and CYP17 A2/A2 genotype were positively associated with the susceptibility of breast cancer.

Objective To construct the KU80 inhibition cell model by RNAi in U2OS cell and to explore the relationship between the Ku80,telomeres and radiosensitivity in telomerase-negative tumor cells.Methods U2OS cells were transfected with the recombinant plasmids of pshRNA-K80 by the lipofectamine,and the stable transfected cell clones were selected by G418.After the selection,the cells were collected and analyzed by the flow cytometry.RT-PCR and Western blot were used to measure the expression of Ku80 and Real-time PCR was used to detect the length of telomeres.The radiosensitivity of U2OS was determined by clone formation array.Results The transfection efficiency of the positive cell clones detected by the flow cytometry was (83.23 ± 7.63) ％.The inhibition rate of the Ku80 gene transcription in the cell group with recombinant plasmid was(68.09 ± 1.16)％ and the inhibition rate of the Ku80 protein expression in the same group was (11.03 ± 2.45) ％.The results of Real-time PCR showed that the telomere length of the cell group with recombinant plasmid (1.07 ± 0.07) was significantly shorter than that of the control group (4.42 ± 1.30,F =38.58,P ＜ 0.05) and that of the empty plasmid group (4.11 ±0.84,F =38.58,P ＜ 0.05).Compared to the control group,the telomere length of the empty plasmid group did not changed(4.42 ±0.84 vs.4.11 ±0.84).U2OS cells with Ku80 expression suppressed had lower SF2 than that of the control cells (F =1089.61,P ＜0.05),and resulted in the SER of 1.47.Conclusions The Ku80 inhibition cell model in telomerase-negative U2OS cell line is successfully established which has the shorter telomere length,and is more sensitive to radiation.Telomere shortening caused by pshRNA-of Ku80 is likely to be one of the mechanisms of radiosensitization in this kind of cell model.