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.

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.

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 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 investigate the efficacy of double-“ U” embedding and pursestring suture and binding pancreaticojejunostomy for the prevention of pancreatic fistula.Methods The retrospective cohort study was adopted.The clinical data of 208 patients who underwent pancreaticojejunostomy at the Hunan Provincial People's Hospital from March 2011 to March 2015 were collected.Of 208 patients,106 patients undergoing double-“ U” embedding and pursestring suture and binding pancreaticojejunostomy were allocated into the double-“ U” group and 102 patients undergoing Child pancreaticojejunostomy were allocated into the Child group.Observation indicators included (1) surgical effects:anastomosis time,postoperative pancreatic leakage,duration of hospital stay,(2) follow-up situations.The follow-up using telephone interview and outpatient examination was performed to detect postoperative long-term complications and recovery of patients by abdominal ultrasound or computed tomography (CT) at every 6 months postoperatively up to September 2015.Measurement data with normal distribution were represented as x ± s and comparison between groups was analyzed by t test.Count data were analyzed using the chi-square test.Results (1) Surgical effects:208 patients underwent successful surgery without occurrence of death.The anastomosis time was (13.0 ± 1.5) minutes in the double-“ U” group and (20.0 ± 1.6) minutes in the Child group,with a statistically significant difference between the 2 groups (t =4.713,P ＜ 0.05).Two patients in the double-“ U” group were complicated with grade A of pancreatic leakage,including 1 of 36 patients with normal pancreatic remnant and 1 of 70 patients with fibrotic pancreatic remnant.Nine patients in the Child group were complicated with pancreatic leakage,including 6 in grade A,1 in grade B and 2 in grade C,and there were 6 of 33 patients (4 in grade A,1 in grade B,1 in grade C) with normal pancreatic remnant and 3 of 69 patients (2 in grade A,1 in grade C) with fibrotic pancreatic remnant.There were statistically significant differences in the pancreatic leakage between the 2 groups and among the patients with normal pancreatic remnant in the 2 groups (x2 =2.951,4.994,P ＜ 0.05).The duration of postoperative hospital stay was (13.5 ± 1.2)days in the double-“U” group and (15.7 ± 2.6)days in the Child group,with a statistically significant difference (t =1.011,P ＜ 0.05).No readmission in the 2 groups occurred.(2) Followup situations:91 of 106 patients in the double-“U” group were followed up for 6-54 months with a median time of 30 months.During the follow-up,8 patients were dead,12 patients didn't undergo reoperation due to multiple metastases in the liver,lung and greater omentum,4 and 4 patients were respectively complicated with relapsing pancreatitis and refluxing cholangitis,and other patients had good conditions without the occurrence of diabetes,diarrhea,indigestion and hypopancreatism.Eighty-eight of 102 patients in the Child group were followed up for 6-54 months with a median time of 25 months.During the follow-up,10 patients were dead,11 patients didn't undergo reoperation due to multiple metastases in the liver,lung and greater omentum,6 and 6 patients were respectively complicated with relapsing pancreatitis and refluxing cholangitis,and other patients had good conditions without the occurrence of diabetes,diarrhea,indigestion and hypopancreatism.Conclusion Double“U” embedding and pursestring suture and binding pancreaticojejunostomy for the prevention of pancreatic fistula can reduce the suture time,incidence of pancreatic leakage and duration of postoperative hospital stay,and it is especially suitable for the patients with normal pancreatic remnant.

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 detect specific cell killing effect of radiation combined with horseradish peroxidase (HRP)/indole-3-acetic (IAA) suicide gene therapy controlled by a novel radio-inducible and cancer-specific chimeric gene promoter in lung cancer. Methods We constructed a plasmid expressing HRP enzyme under the control of chimeric human telomerase reverse transcriptase (hTERT) promoter carrying 6 CArG elements, a plasmid expressing HRP enzyme under the control of hTERT promoter carrying single CArG element, and two control plasmids, which named pE6-hTERT-HRP, phTERT-HRP, pControl-HRP, and pControl-luc, respectively. After radiation, the proliferation inhibition and apoptosis induction effect of each type of plasmid in lung cancer cells (A549, SPC-A1) and normal lung cells (hEL) was detected by cell counting and Annexin V-FITC staining. The change of radiosensitivity of lung cancer cells with plasmid system was also detected by clonogenic assays. Results After a single dose radiation of 6 Gy,the average proliferation inhibition rates of pE6-hTERT-HRP, phTERT-HRP, pControl-HRP, and pControl-luc systems were 72. 92% ,40.60% , 51.00% and 25.19% (F= 67.31 , P< 0.01) in A549 cells ,64.63%,30.02%,48.23% and 23.16% (F=64.94, P< 0.01) in SPC-A1 cells, and 20.81%,18.05%, 44.20% and 18.32% (F=52. 19,P<0.01) in normal hEL cells, respectively. The average early apoptosis rates of these four plasmid systems were 36. 63%, 22. 30%, 24. 33% and 12. 53% (F =50. 99,P <0. 01) in A549 cells, 33.73%, 17. 37%, 22. 43% and 11.20% (F = 20. 76, P < 0. 01) in SPC-A1 cells, and 13.53 %, 12. 5%, 21.93% and 12. 16% (F = 15. 08, P < 0. 01) in normal hEL cells,respectively. The sensitizing enhancement ratios of the four plasmid systems were 3.45, 2. 29, 3.05 and 1.21 in A549 cells, while 2. 68, 2. 15, 3.05 and 1.21 in SPC-A1 cells, respectively. Conclusions The new suicide gene system controlled by chimeric promoter may provide a novel therapeutic modality for lung cancer.

Immunohistochemistry was used to detect the protein expression of Snail in 5 specimens jusxta-eancerous normal breast tissues, 35 specimens of cancerous tissues without metastasis and 20 specimens of breast carcinoma with lymphonode metastasis. Breast carcinoma cell line MDA-MB-231 was transfected with antisense Snail. Results showed that the expression of Snail protein was significantly higher in breast carcinomas than in their normal tissues. The mRNA and protein expressions of Snail in the breast carcinoma cells treated with antisense Snail was significantly decreased while the E-cadherin protein significantly increased (both P < 0.05). The number of invasive ceils treated with antisense Snail was (10.5±1.3)%, while in treated with EGFP was (68.2±2.1)% (P < 0.05). The abnormal expression of Snail contribute to the invasiveness of breast carcinoma. The antisense Snail could prevent the cells ability to invade in vitro, and the effect is related with the up-regulated E-cadherin protein.