Objective:To explore the expression of Netrin-1 protein in human renal clear cell carcinoma (RCCC) and the relationships between Netrin-1, pathology and prognosis. Methods:72 cases of RCCC admitted in our hospital from 2008 June to 2009 June and their adjacent tissues were selected for study. They included 30 cases in stage I-II, 42 cases in stageⅢ-IV;9 cases in grade I, 9 cases in grade II, 40 cases in gradeⅢand 14 cases in grade IV. All cases were followed up for more than 5 years. Survival analysis lines were made by Kaplan—Meier method and the difference between groups was tested by the Log-rank test. The expression of Netrin-1 protein was detected by immunohistochemistry staining and its clinical significance was analyzed. Results:Renal clear cell carcinoma:51 cases in high expression of Netrin-1 and 21 cases in low expression, normal tissues: 12 cases in high expression of Netrin-1 and 60 cases in low expression, the difference between the two groups is significant (χ2=42.921, P<0.01). The difference of the expression of Netrin-1in Fuhrman grade and AJCC clinical stage is significant (χ2=8.000,χ2=6.203;P<0.05). The 5-year survival rate in low protein expression group and in high protein expression group was 79%(17/21) and 62%(32/51). The survival curve had different trend, with no significant difference between groups (χ2=1.360, P=0.245). Conclusions: Netrin-1 protein plays an important role in the development of RCCC. It might be a new specific tumor marker of RCCC, and might become a new target in treatment of RCCC.

Objective To develop a blooding device for blood sucking insects. It's designed for researches, such as artificial breeding of indoor medium insects, evaluation of vermicides'effect, and artificial infection of pathogenic microorganisms. Methods This device was developed with AT89C51 MCU. Results The desired instrument could provide blood which could be adjusted from 30℃to 38℃. The error of temperature could be controlled within 0.5℃. Conclusion The instrument is easily-operated and stable. It can replace the artificial blood-breeding.

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Objective: To investigate the effects of VPA (valproate) on proliferation, cell cycle distribution and apoptosis of human kidney carcinoma ACHN cells and the possible underlying mechanisms. Methods: The effect of VPA on the proliferation of ACHN cells was examined by CCK-8 (cell counting kit-8) assay. Flow cytometry was used to analyze the cell cycle distribution and apoptosis of ACHN cells exposed to VPA. The mRNA expressions of cyclin E1, P 21WAF1, Bcl-2 and Bax were detected by real-time fluorescence quantitative-PCR. Results: Incubation with various concentrations of VPA for 48 h resulted in a significant inhibition of proliferation of ACHN cells with an IC50 (50% inhibitory concentration) value of 4.21 mmol/L. After treatment with VPA, the cell cycle was arrested obviously at G 0/G1 phase and the apoptotic rate was significantly increased as compared with the control group. After treatment with 4 mmol/L VPA for 48 h, the levels of P21WAF1 and Bax mRNAs were both significantly increased, and at the same time, the levels of cyclin E1 and Bcl-2 mRNAs were obviously decreased. Conclusion: VPA can inhibit the proliferation of kidney carcinoma ACHN cells by inducing cell-cycle arrest and apoptosis. Copyright © 2013 by TUMOR.

Objective To detect neuroendocrinal factor,and explore whether neurohormonal activation exists in children with conge-nital heart disease(CHD).Methods Concentrations of atrial natriuretic peptide(ANP),brain natriuretic peptide(BNP),angiotensionⅡ(AngⅡ),aldosterone(ALD),endothelin-1(ET-1),and norepinephrine(NE)were determined in 100 children with CHD and were assayed at 24 hours,1 month,3 months and 6 months after interventional therapy in 70 cases with CHD.Results Children with CHD heart disease had elevated levels of ANP(25.6?7.5) pmol/L,BNP(15.7?7.4) pmol/L,ET-1(1.12?0.31) pmol/L(all P=0.0001),(AngⅡ)(90.3?11.5) ng/L,NE(1.07?0.08) nmol/L and ALD(246.1?42.3) pmol/L(all P=0.001).There was a highly significant stepwise increase in ANP,BNP,ET-1,AngⅡ,ALD and NE according to New York Heart Association class,with even asymptomatic patients having evidence of significant neurohormonal activation.The neurohormonal function gradually returned to normal after interven-(tional) therapy.Conclusions Neurohormonal activation in children CHD bears the hallmarks of chronic heart failure,relating to symptom severity and ventricular dysfunction.

Objective To synthesize 628F-Py-AMD3465,to investigate its biodistribution in mice and to perform the microPET/CT imaging on mice bearing human lung cancer cell (A549).Methods AMD3465 quaternary ammonium salt precursor was directly labeled with 18F,then 628F-Py-AMD3465 was synthesized through nucleophilic reaction,hydrolysis,neutralization and the product was purified using HPLC.The labeling yield and radiochemical purity were analyzed by HPLC.Fifteen Kunming mice were injected with 5.55 MBq of 628F-Py-AMD3465 and sacrificed at 5,20,40,60 and 120 min postinjection.The selected tissues were harvested and weighed,and the radioactivity in the tissues was measured by an automated γ-spectrometer.The ％ID/g was calculated.MicroPET/CT studies were performed on A549-bearing mice after injecting 6-18F-Py-AMD3465 through vena caudal.Paired t test was used.Results 6-18F-Py-AMD3465 was successfully synthesized with the labeling yield of (9.0±2.0)％,the total synthesis time was about 60 min,and the radiochemical purity was more than 98％.Biodistribution studies showed that the radiouptake was higher in the kidneys and bladder of normal mice,which demonstrated that 6-18 F-Py-AMD3465 was mainly excreted through the kidneys.Biodistribution in A549-bearing mice was similar to that in normal mice.The tumor/muscle ratio at 40 min was 5.0,but the radiouptake of the tumor was still lower than that of the normal lung:(8.05±0.35) ％ID/g vs (9.33±0.66) ％ID/g;t=5.26,P＜0.05.MicroPET/CT imaging showed that the high-uptake location of 6-18F-Py-AMD3465 in tumor-bearing mice was similar to the normal mice,and the tumor uptake reached the maximum level at 45 min post-injection (SUV 0.67).Conclusions 6-18F-Py-AMD3465 can be synthesized by a simple method.A lower uptake could be shown in the tumor compared to that in the lung and the tracer has limited diagnostic value for lung cancer.

Objective To establish a method for the determination of gastrodin in Tianyuan Granula. Methods The sample was refluxed with methanol, then eluted with 80 % ethanol through the alumina column(neutral). The chromato-graphic conditions were as follows: Diamonsil Cts chromatographic column(250 mm ×4.6 mm, 5 μm) with a mobile phase of acetonitrile-0. 15 % phosphoric acid (1 : 99), the detection wavelength being at 220 nm and the flow rate be-good (r=0.9999 95, n=7).The average recovery was 99.09 % and RSD=1. 50 % (n=6). Conclusion The method for determination of gastrodin in Tianyuan Granula is accurate, and with a good reproducibility.

Objective To improve the accuracy of diagnosis of the greater omentum blunt trauma(GOBT)by analyzing the MSCT findings and pathological features.Methods The MSCT appearances of 5 1 cases of GOBT with operation and (or)pathological con-firmed were analyzed retrospectively.All patients were underwent MSCT plain scan in whole abdomen.Two experienced radiolo-gists blinded to the clinical diagnosis,reached a diagnosis from the MSCT findings by two ways of plain and plain joint post-process-ing MSCT images.The detection rates of GOBT were compared by two-related-samples Fisher exact probability chi-square test.Re-sults Fifty-one cases of GOBT were distributed over the intestine,in the superior abdominal cavity,the left and the right abdominal cavity,those cases were 27,18,3 and 3 respectively.The MSCT findings of GOBT mainly consist of greater omentum(GO)streak-ing,GO hematoma and fluid around GO.Those detectable rates were 80.3%,68.6%and 90.2 % respectively.The accuracy in di-agnosis of GOBT with MSCT was 94.1%(48/51)in our study.The detection rate of GOBT by two ways was significant (P=0.04).The detection rate of GOBT was increased obviously by the way of plain joint post-processing MSCT images.Conclusion MSCT manifestations of GOBT has some characteristics.A more accurate diagnosis of GOBT could be made according to the MSCT findings by plain joint post-processing MSCT images.

Objective: To express glycosyl-phosphatidylinositol (GPI) modified Met- RANTES fusion protein on Chinese hamster ovary (CHO) cells and to develop a novel immunosuppressant GPI anchored form of Met-RANTES. Methods: The eukaryotic expression vector PEF/GPI-Met-RANTES were constructed and transfected into CHO cells by electroporation. The transfectants were selected with methotrexate (MTX). Expression of the recombinant protein was assessed by flow cytometric analysis, cell immunofluorescence staining and immunogold electron microscopy. Results: The chimeric molecules of GPI anchored form of Met-RANTES including the whole reading frame were constructed, and the sequence was identical to the designed sequence. GPI anchored form of Met-RANTES was stably expressed on CHO- DHFR- cells. Conclusion: A large amount of GPI modified Met-RANTES fusion protein was expressed on CHO cells. GPI anchored form of Met-RANTES may be used as novel immunosuppressant for suppressing reaction in graft rejection.

ObjectiveTo study the feasibility of ex vivo gene transfer to donor heart by delivering i ntracoronarily a reporter gene (Lac-Z gene) to donor heart at the time of trans plantation.MethodsThe model of heterotopic heart transplantation in murine was established and the method of perfusing intracoronarily was performed. Twelve male BALB/c mice were divided into 2 groups randomly: gene-transfer group (group 1) and control grou p (group 2). In group 1, plasmid vector (PSV-?-gal containing Lac-Z gene)/li posome (DOSPER) was perfused intracoronarily into each donor heart at the time o f transplantation. In group 2, normal saline was perfused. Donor hearts were har vested 3 days after transplantation. Freezing sections were made for detection o f the transfer and expression of Lac-Z gene by histochemical staining (X-gal). ResultsThe expression of Lac-Z gene was detected in the donor hearts of group 1. In tw o donor hearts, the expression of Lac-Z gene was detectable in the myocardial c ells in the mid-layer of ventriculus; In one donor heart the expression was fou nd in the myocardial cells under epicardium. But no expression of Lac-Z gene wa s detected in donor hearts of group 2.ConclusionEx vivo gene transfer intracoronarily to donor heart in the form of plasmid vector /liposome complex at the time of transplant ation is feasible.