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.

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.

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 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.

Objective To detect the selective inhibitory effects of irradiation plus adenovirusmediated horseradish peroxidase ( HRP)/indole-3-acetic acid (IAA) suicide gene system using tumorspecific and radio-inducible chimeric promoter on human hepatocellular carcinoma subcutaneously xenografted in nude mouse.MethodsRecombinant replicated-deficient adenovirus vector containing HRP gene and chimeric human telomerase reverse transcriptase (hTERT) promoter carrying 6 radioinducible CArG elements was constructed.A human subcutaneous transplanting hepatocellular carcinoma (MHCC97 cell line) model was treated with -γ-ray irradiation plus intra-tumor injections of adenoviral vector and intra-peritoneal injections of prodrug IAA.The change of tumor volume and tumor growth inhibiting rate,the survival time of nude mice,as well as histopathology of xenograft tumor and normal tissues were evaluated.Results Thirty one days after the treatment,the relative tumor volumes in the negative,adenovirus therapy,irradiation,and combination groups were 49.23 ± 4.55,27.71 :± 7.74,28.53 + 10.48 and 11.58 ± 3.23,respectively.There was a significantly statistical difference among them (F = 16.288,P ＜0.01 ).The inhibition effect in the combination group was strongest as compared with that in other groups,and its inhibition ratio was 76.5%.The survival period extended to 43 d in the combination group,which showed a significantly difference with that in the control group(x2 = 18.307 ,P ＜0.01 ).The area of tumors necrosis in the combination group was larger than that in the other groups,and the normal tissues showed no treatment-related toxic effect in all groups.However,multiple hepatocellular carcinoma metastases were observed in the liver in the control group,there were a few metastases in the monotherapy groups and no metastasis in the combination group.Conclusions Adenovirus-mediated suicide gene therapy plus radiotherapy dramatically could inhibit tumor growth and prolong median survival time.It might provide a promising therapeutic modality for hepatocellular carcinoma therapy.

Objective To investigate the therapeutic effects and toxic effects of radiochemotherapy versus chemotherapy alone in patients with advanced gastric cancer after gastrectomy.Methods A total of 183 patients with stage Ⅱ-Ⅳ (M0) gastric cancer,who received treatment after gastrectomy from 2004 to 2010,were included in the study.Of the 183 patients,64 received radiochemotherapy after gastrectomy,and 119 received chemotherapy alone after gastrectomy.The survival rates and hematological and gastrointestinal toxic effects were compared between the two groups;survival difference was also analyzed after the patients were stratified by TNM stage,number of metastatic lymph nodes,and extent of lymph node dissection (D0,D1,and D2).The Kaplan-Meier method was used for calculating survival rates,and the log-rank test was used for survival difference analysis and univariate prognostic analysis;the chi-square test was used for comparing toxic effects between the two groups.Results The follow-up rate was 87.8％ ;72 patients were followed up for at least 3 years,and 29 patients for at least 5 years.The 1-,3-,and 5-year survival rates for the radiochemotherapy group were 86％,62％,and 55％,respectively,as compared with 77％,53％,and 43％ for the chemotherapy group (P =0.079).There were no significant differences in grade 3-4 hematological and gastrointestinal toxic effects between the two groups (P =0.363 and 0.617).The univariate analysis showed that radiochemotherapy had a significantly better survival benefit than chemotherapy alone in patients with stage ⅢB-Ⅳ (M0) gastric cancer,patients who underwent D0 lymph node dissection,and patients with more than 6 metastatic lymph nodes (P =0.022,0.025,and 0.021).Conclusions Compared with chemotherapy alone,radiochemotherapy tends to improve survival in patients with gastric cancer after gastrectomy,and its toxic effects are tolerable.The patients with stage ⅢB-Ⅳ (M0) gastric cancer,patients who undergo D0 lymph node dissection,and patients with more than 6 metastatic lymph nodes can benefit from radiochemotherapy.

BACKGROUND:Currently, cellular composition and the features of the nucleus pulposus are stil not to be clarified.
OBJECTIVE:To establish the in vitro culture system of rat nucleus pulposus-derived mesenchymal stem cells and to identify their multi-lineage differentiation potential.
METHODS:Mesenchymal stem cells from the nucleus pulposus tissues of Sprague-Dawley rats were cultured in vitro. Then, cells at passage 3 were induced to differentiate into osteoblasts, adipocytes and chondrocytes as experimental group. cells cultured with basic culture medium served as controls.
RESULTS AND CONCLUSION:cells isolated from rat nucleus pulposus could form the sunflower-like colonies and exhibit clone-like growth when they cultured at a low density. cells at passage 3 became homogeneous and exhibited fibroblast-like morphology. After 28 days of osteogenic induction, arizarin red positive signals were detected in the experimental group. The mRNA expressions of RunX2, osteopontin and osteocalcin were significantly increased in the experimental group, compared to the control group (P<0.05). After 21 days of adipogenic induction, oil red-O positive cells were detected in the experimental group. The mRNA expressions of C/EBPαand PPARγ2 were significantly increased in the experimental group, compared to the control group (P<0.05). After 21 days of chondrogenic induction, safranin O/fast green staining was positive in the experimental group. The mRNA expressions of aggrecan and Col2a1 were significantly increased in the experimental group, compared to the control group (P<0.05). Our findings in this study suggested that nucleus pulposus-derived mesenchymal stem cells could be isolated from the Sprague-Dawley rat nucleus pulposus and exhibited clonal-like growth when they were cultured in vitro. These cells were confirmed to have the potential to differentiate into adipocytes, osteoblasts and chondrocytes in vitro.

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.