Objective To construct a stable expression plasmids miR-218,to lay the experimental foundation for the expression and mechanism of miR-218 in human renal cell carcinoma.Methods The primers were designed by Has-miR-218 and the miR-218 fragment were amplified by PCR,and then which was connected with the carrier pcDNA3.1 (+) to build a stable expression plasmids.The recombinant plasmids were trasfected into renal cell carcinoma cell line 769-P,and the expression of miR-218 in these cells were detected.Results The plasmids were constructat successfully,which was determind by enzyme digestion and sequencing results.The constructed expression plasmids pcDNA3.1-miR-218 transfection of human renal cell carcinoma cell line 769-P,miR-218 cxpression level of cells was significantly increased after trasfected by recombinant plasmids carrying miR-218 gene (relative expression quantity was 1.64).Conclusion The miR-218 expression plasmids pcDNA3.1-miR-218 were successfully constructed,the plasmids can be applied to the study of miR-218 function and mechanism in renal cell carcinoma.

Objective To investigate the association of inducible nitric oxide synthetase (iNOS) expression with the expression of mutation type p53 (mt p53) in prostatic carcinoma (PCa). Methods The expression levels of iNOS and mt p53 were in 28 PCa specimens and 10 cases of benign prostatic hyperplasia (BPH) were detected by immunohistochemical method. Results The expression levels of both iNOS and mt p53 in PCa were higher than those in BPH. The expression of iNOS was related to the expression of p53 protein. Conclusion iNOS and mt p53 may play a role in the pathogenesis of PCa.

Child ; China ; Humans ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; diagnosis ; therapy

Objective To investigate effects of Kanglaite injection on proliferation, cycle and apoptosis of human breast cancer MCF-7 cells;To discuss its relevant mechanism. Methods Logarithmic growth phase cells were divided into control group and Kanglaite-treatment group (10, 20, 40μL/mL). Cells were cultured in RPMI-1640 for 24 h before drug treatment. The inhibition rate of Kanglaite injection on proliferation of human breast cancer MCF-7 cells was detected by MTT assay. Apoptosis and cell cycle of MCF-7 cells were detected by flow cytometry. Changes in cell nucleus were determined by Hochest staining assay. Protein expressions of Bcl-2 and Bax were detected by ELISA and Western blot. Results Kanglaite injection for 12 h, 24 h or 48 h resulted in a significant inhibition of MCF-7 cells proliferation (P<0.05, P<0.01);Compared with the control group, Kanglaite injection-treated cells showed increased percentage in G2/M and G0/G1 phases (P<0.001, P<0.01), but showed decreased percentage in S phase (P<0.01), and apoptosis rate increased (P<0.05, P<0.001). Kanglaite injection significantly decreased protein expression of Bcl-2, and enhanced protein expression of Bax of MCF-7 cells (P<0.01, P<0.001). Conclusion Kanglaite injection can inhibit the proliferation of human breast cancer MCF-7 cells, decrease cell cycle and induce apoptosis, the mechanism is related with decreasing protein expression of Bcl-2 and enhance the protein expression of Bax.

Objective To discuss prevention and treatment of the late complications of deteniaceacal-colon continent urinary reservoir. Methods 105 patients who underwent detenia ceacal-colon con-tinent urinary reservoir were followed up for 9 -82 months(mean 40.5 months).Clinical data of these wereretrospectively analyzed. Results Late complications occurred in 25.7%(27/105) of the cases and 9were treated with operations,including difficulty with catheterization in 4 cases(3.8%),3 treated with dila-tion and 1 with efferent limb reconstruction with ileal,urinary pouch stones in 5(4.8%),4 treated with li-thotomy,urinary pouch perfotation in 1 (0.95%),treated with repair,urinary pouch superdistention in 1(0.95%),treated with reduction, single or double lateral hydronephrosis in 8(7.6%),caused by strictureof single or double lateral lower ureters in 4 (2 treated with excision lesion and anastomosis ) and by refluxin 1,1 case suffering from azotemia preoperatively worsened to uraemia treated with regular haemodialysis,hyperchloremia in 7 (6.7%),only 1 needed drugs treatment,repeated urinary infection in 11,amomg them,10 were complicated by the other complications,all treated with sensitive antibiotic. Conclusions Most ofpatients underwent detenia ceacal-colon continent urinary reservoir had good curative effects. Late complica-tions afflicting these patients mostly had immediate or mediate relations with symptomatic urinary infection.Prevention and treatment of symptomatic urinary infection can reduce the occurrence of late complications ofthis operative method,regular and adequant pouch washing and catheterization is efficient to prevention oflate complications of this operative method.

Objective To explore the effects of miR-218 on biological functions of renal cell carcinoma cell line through regulating the expression levels of miR-218.Methods The pcDNA3.1-miR-218 was transfected into renal carcinoma cell line A498 and 769-P and the expression of miR-218 was detected.The cell activity,invasion,apoptosis and proliferation of transfected renal carcinoma cell line were analyzed in vitro.Results After plasmid transfected A498/769-P renal carcinoma cell line,the expression level of miR-218 (1.99,1.64) was significantly higher than that of the control group (1.00) (t =60.82,10.89,P ＜ 0.000 1) The cell viability (0.90±0.10,0.68±0.06) was lower than that of the control group (1.39±0.14,1.24±0.08) by CCK8 experiment (t =15.02,31.69,P ＜ 0.000 1).The cell invasion was lower than that of control group by Transwell experiment (t =15.78,18.80,P ＜ 0.000 1).The apoptosis ratio was higher by AnnxinV-FITC experiment,the apoptosis ration of control group and transfected group were respectively 0.25 ％,45.77 ％ in A498 cell line and 0.11％,45.57 ％ in 769-P cell line.The proliferation was lower than that of the control group (P ＜ 0.000 1).Conclusion Up-regulation of miR-218 expression can inhibit the growth of renal cell carcinoma cell line in vitro.

Objective To identify epitope of neutrophil-activating protein(NAP) of Helicobacter pylori(Hp).Methods Using the mouse monoclonal antibodies against NAP as selective molecular and immunoscrcening phage-display random 7-peptides library.The positive clones were sequenced and analyzed.Phage clones were chosen to immunize mice and to evaluate the potential of phagotopes as effective vaccines.Results One mimotope(FAHLATQ)showed a good match with the NAP at 140-143 AA(AHLA)and the serum of mice induced by the phage clone clearly recognized NAP.Conclusion This study suggests thatthe antigenic epitope could be mapped through screening the phage-display peptide library with monoclonalantibody and a mimotopo of NAP providing an ahernative approach for the diagnosis and development of avaccine for Hp.

Objective To explore the application of high-frequency oscillatory ventilation(HFOV)postoperatively after open heart surgery in children with congenital heart disease.Methods From Jun.2007 to Jun.2008,a total of 1 957 cases were operated in pediatric center of Fuwai cardiovascular hospital.Thirteen cases of 1 957 were given HFOV.Among these 13 patients,8 were male and 5 were female;Age was(13.4?15.4)months.The range of weight was 4.2-12.5 kg.Four cases were tetralogy of Fallot,3 cases were transposition of the great arteries(1 case was corrective transposition of the great arteries),2 cases were total anomalous pulmonary venous connection,2 cases were total endocardial cushion defects,2 cases were pulmonary artery atresia,all patients were under cardiopulmonary bypass repaired operation except 1 patient with PAA under Blalock-Taussig procedure.All patients were mechanical ventilated after surgery.Results The duration of switching general mechanical ventilation to HFOV was(15.70?20.33)days and patients continued to be on HFOV about(10.0?7.5)days.In short term,the lung function of ventilation and gas exchange were be improved by HFOV,but the long-term result was not sa-tisfactory due to a high mortality.There were 5 survived(38.4%)in this group.Six cases of the patients succumbed to consolidation of lung.The commonest complication was pneumothorax(7 cases).Conclusions Short term of HFOV in children with congenital heart disease was satisfactory postoperatively.But there was a high mortality.HFOV can be regarded as a transition method for unsatisfactory long-term.

Objective To establish the reference databases for bone mineral density(BMD)in multiple skeletal regions,which would be useful for diagnosis of osteoporosis(OP)and prediction of fracture risk in adult women in Qingdao.Methods BMD was measured by dual-energy X-ray absorptiometry at skeletal regions of lumbar spine,left hip(femoral neck,Ward's triangle and greater trochanter)in 868 healthy adult women aged 25- 83 years and 191 women with fractures.BMD of skeletal regions with age-related change was found to fit in 8 kinds of regression models.Best model equations of fitting were found and the reference database was established.BMD of women with fractures was compared with the reference databases to predict the risk of fracture.Results BMD in 6 skeletal regions changed with aging and a cubic regression model was better fitted with aged-related change as compared with other regression models.The coefficients of determination(R~2)of fitting curve were 0.21?0.09 (P＜0.01).The BMD reference databases of women in Qingdao were established by cubic regression equation, the peak BMD of lumbar spine and hip appeared at 25-29,and 40-44 years old groups respectively.Finally,the BMD in fracture group was significantly decreased by 1.6-2.5 s as compared with the peak BMD of health women. Conclusion The bone quantity is lost rapidly after 45 years old in women.If the BMD in women after 50 years old is decreased by 1.6-2.5 s compared with the peak BMD in the same area,the risk of fracture is increased.

The content of benzyl isothiocyanate (BITC) which as the enzymatic hydrolysis product of benzyl glucosinolate through thioglucosidase was determined by HPLC. The content of benzyl isothiocyanate (BITC) which as the enzymatic hydrolysis product of benzyl glucosinolate through thioglucosidase was determined by HPLC. The chromatography condition was as follows: Kaseisorb LC ODS 2000 (4.6 mm x 150 mm, 5 min) column with the mobile phase of acetonitrile(A)-water( B) under gradient elution (0-5 min, 3%-8% A; 5-9 min, 8%-48% A; 9-23 min, 48%-62% A; 23-28 min, 62%-99% A); the flow rate was 1.0 mL x min(-1) with 10 microL injection volume; detection wavelength was 246 nm and temperature of column was 40 degrees C. The content of benzyl glucosinolate was in the range of 10.76-17.91 g x L(-1). The method is simple, accurate and good reproducibility which can be used for the determination of benzyl glucosinolate in Lepidium meyenii, effectively.
Chromatography, High Pressure Liquid ; methods ; Glucosinolates ; analysis ; Lepidium ; chemistry ; Plant Extracts ; analysis