Expression of dead box 1 gene in neuroblastoma / 中华实用儿科临床杂志

Objective To detect the expression of dead box 1(DDX1)gene in tumor tissue and pericarcino-matous tissue of clinical neuroblastoma(NB)samples,and explore the relationship between DDX1 and NB. Methods Five cases of pathological specimens in children with NB were chosen from Department of Pathology,the First Affi-liated Hospital of Xinxiang Medical University between January 2012 and December 2014. In the 5 cases,3 cases were male,2 cases were female,the age of 1 - 5 years old,average age(2. 1 ± 1. 6)years. The NB tissue and pericarcino-matous tissue(pericarcinomatous tissue was normal tissue which was at least 2 cm from the tumor tissue)of 5 children were collected and fixed in 40 g/ L formaldehyde solution. Then with the conventional dehydration,embedding,sectio-ning, dewaxing, hydration, antigen repair, add primary antibodies, secondary antibodies, diaminobenzine chromogenic. The expressions of DDX1 in tumor tissue and pericarcinomatous were observed with light microscopy and with semi - quantitative analysis.(1)Staining degree:no staining with 0 score;light staining with 1 score;medium staining with 2 scores;deeply staining with 3 scores.(2)Positive cells proportion:positive cells proportion ﹤ 10% with 0 score;positive cells proportion within 10% - 30% with 1 score;positive cells proportion within 31% - 60% with 2 scores;positive cells proportion ﹥ 61% with 3 scores. Final scores were a half of the sum of staining degree score and positive cells proportion score,final score within 0 -1. 0 with - ,1. 1 -2. 0 with + ,2. 1 - 3. 0 with + + ,3. 1 -5. 0 with + + + . Results DDX1 were expressed in NB and pericarcinomatous tissues,but visible DDX1 positive staining number more and deeper in NB,DDX1 positive staining number less and light in pericarcinomatous tissues. Five cases of pericarcinomatous tissues immunohistochemical semi - quantitative score were negative and final scores were 1. 0 score or less,the mean value was 0. 5 score. NB immunohistochemical semi - quantitative score were+or + +and final scores were 1. 5 score or higher,the mean value was 1. 8 scores,the expressions of DDX1 were sig-nificantly higher in NB than the pericarcinomatous tissues. Conclusions DDX1 is highly expressed in NB,which may contribute to the development of NB. This suggest DDX1 may serve as an oncogene and play a catalytic role in the de-velopment of NB,which provides a clinical evidence for the follow - up study.

Effects of dead box 1 gene on invasion, migration and drug resistance capability of neuroblastoma cells / 中华实用儿科临床杂志

Objective To explore the effects of dead box 1 (DDX1) gene on invasion,migration and drug resistance capability of neuroblastoma(NB) cells.Methods According to the virus drop degree,the appropriate amount of target virus(Lenti-DDX1-MIR virus liquid,drop degrees 1012 TU/L) and negative control virus(Lenti-EGFP virus liquid,drop degrees 3 × 1011 TU/L) (multiplicity of infection was 10) were added into 2 hole cells,respectively.SK-N-BE(2)/blank,SK-N-BE(2)/shV and SK-N-BE(2)/shDDX1 cells which grew in good condition were cultured.Transwell chamber was used to detect the invasion,and cell staining was made with crystal violet.The researchers calculate 5 field counting in each small room and calculate the average cell invasion rate.Transwell chamber was used to detect the migration,and cell staining was made with crystal violet.The 570 nm absorbance values was tested with enzyme linked immunosorbent assay (ELISA) reader,and cell migration was calculated.The researchers used 50 mg Cisplatin,solution 10 g/L mother liquor standby with 5 mL dimethyl sulphoxide,and 50 mg Doxorubicin,solution 1 g/L mother liquor standby with PBS.Drugs were added to the cell culture plate,and Doxorubicin final concentration was 1.0 mg/L,and Cisplatin final concentration was 2.5 mg/L,and photographic record was documented after drug treatment for 24 h.Cell Count Kit-8 (CCK-8) was used to detect the drug sensitivity to NB cells to Doxorubicin and Cisplatin.Results Transwell results showed that,cell invasion concentration in SK-N-BE (2)/shDDX1 was 60％ compared with SK-N-BE (2)/blank and SK-N-BE (2) /shV;Crystal violet staining showed that cell invasion of SK-N-BE(2)/shDDX1 was significantly weaker than that of SK-N-BE(2)/blank and SK-N-BE(2)/shV cells,that is to say,DDX1 knockdown reduced the cell invasion of NB.Transwell results showed that,cell migration concentration in SK-N-BE(2)/shDDX1 was 50％ compared with SK-N-BE(2)/blank and SK-N-BE(2)/shY;Crystal violet staining showed that cell migration of SK-N-BE(2)/shDDX1 was significantly weaker than that of SK-N-BE(2)/blank and SK-N-BE(2)/shV cells,that is to say,DDX1 knockdown reduced the cell migration of NB.With DDX1 knockdown,24-h inhibition rate of SK-N-BE (2)/shDDX1 cell was 1.93 times of SK-N-BE (2)/shV cell with 1.0 mg/L Doxorubicin,24 h inhibition rate of SK-N-BE(2)/shDDX1 cell was 1.38 times of SK-N-BE(2)/shV cell with 2.5 mg/L Cisplatin.DDX1 knockdown could increase the Doxorubicin and Cisplatin drug sensitivity to NB cells.Conclusion DDX1 knockdown can decrease the cell invasion,migration and resistance capability of NB and increase the Doxorubicin and Cisplatin drug sensitivity of NB cells.

Effects of dead box 1 gene on cell apoptosis, proliferation and cell cycle of neuroblastoma cells / 中华实用儿科临床杂志

Objective To explore the effects of dead box 1 (DDX1) gene on cell apoptosis,proliferation and cell cycle of neuroblastoma(NB) cells.Methods SK-N-BE(2)/blank,SK-N-BE(2)/shV and SK-N-BE (2)/shDDX1 cells were seeded in 96-well plates,which grew in good condition and in the logarithmic growth phase,5 000 cells were inoculated in each well and 5 repeated holes were set.Cell count kit 8 (CCK-8) was used to detect the cell number at 12 h,24 h,36 h,48 h,and the average was calculated.The time (hour) was set as abscissa,the optimal density (A) value at 450 nm was set as vertical axis,and the growth curves of these 3 cells were drawn to investigate the effects of DDX1 on the proliferation of NB cells.After 24 h,flow cytometry (PI staining) was used to detect the apoptosis of SK-N-BE (2)/blank,SK-N-BE (2)/shV and SK-N-BE (2)/shDDX1 cell lines to observe the effects of DDX1 on the apoptosis of NB cells.After 24 h,flow cytometry (PI staining) was used to detect the proportion of SK-N-BE(2)/blank,SK-N-BE(2)/shV and SK-N-BE(2)/shDDX1 cells at G1,S,M,G2 stage to observe the effects of DDX1 on the cell cycle of NB cells.Results SK-N-BE (2)/shDDX1 cell proliferation was significantly lower than SK-N-BE(2)/blank and SK-N-BE(2)/shV cells,that was to say,DDX1 knockdown reduced the cell proliferation of NB.According to the flow cytometry results,the total average apoptosis rate was 5.28％ in SK-N-BE (2)/shV cells,and the total average apoptosis rate was 9.99 ％ in SK-N-BE (2)/shDDX1 cells.The number of apoptotic SK-N-BE (2)/shDDX1 cells was significantly higher than the number of SK-N-BE (2)/blank and SK-N-BE (2)/shV cells,which indicated that DDX1 knockdown increased tumor cell apoptosis of NB.Compared with SK-N-BE(2)/blank and SK-N-BE(2)/shV cells,the cell cycle of SK-N-BE(2)/shDDX1 cells was arrested,and the proliferation was affected.Conclusions After DDX1 expression is inhibited,the cell cycle of NB cells are affected,the cell apoptosis is increased,and the cell proliferation is reduced.

Non-fermenter isolation and drug resistance analysis of clinical specimens from the year of 2010 to 2012 / 中华实用儿科临床杂志

Objective To study the non-fermenter isolation rate and its drug resistance to more than 10 kinds of antibiotics at the First Affiliated Hospital to Xinxiang Medical University from the year of 2010 to 2012 in order to offer evidences for reasonable use of drugs by doctors and to prevent nonfermenters from becoming common pathogenic bacteria.Methods The infection and drug resistance of different Nonfermenters (Pseudomonas aeruginosa,Acinetobacter baumannii,Pseudomon as mahophilia,Los non Acinetobacter,Pseudomonas stutzeri,Achromobacter,Bacillus steady short,onions Bock cepacia,meningitis septicemia Elizabeth Kim bacteria)during present 3 years were analyzed by using WHONET 5.5 software.Results The infection rate from non-fermenters as pathogenic bacteria was gradually increased,with the rate from 18.2％ to 27.2％,until 31.2％ ; Acinetobacter Baumannii were isolated as higher infection rate and multiple drug resistance (7.1％ to 18.5％ until 28.7％) in pathogenic bacteria; the infection rate of other non-fermenter with infection rate of high and multiple drug resistance in pathogenic bacteria were gradually increased,such as infection rate or the precedence of Achromobacter xylosoxidans and Pseudomonas maltophilia,especially the precedence of Pseudomonas maltophilia ranged the third in non-fermenters,and Pseudomonas maltophiliai had a higher and multiple drug resistance to many antibiotics; infection rate of non-fermenters with low drug resistance gradually decreased,such as Pseudomonas fluorescens; the drug resistance rate of Acinetobacter Baumannii was higher than that of Pseudomonas aeruginosa and its drug resistance rate was over 50％ to common antibiotics used clinically,as well as Pseudomonas maltophiliai and Empedobacter brevis,especially the resistance rate of Pseudomonas maltophiliai to many commonly used antibiotics was more than 90.0％.Conclusions Non-fermenter isolation rate is becoming higher clinically and its drug resistance becomes more severe,so the sanitation administration department should pay more attention to the use of antibiotics in order to prevent and control drug resistance and higher infection rate caused by non-fermenters as a serious consequence.

Effect of dexamethasone on vascular endothelial growth factor in cerebrospinal fluid of rabbits with bacterial meningitis / 中国小儿急救医学

Objective To investigate the effects of dexamethasone (DEX) on vascular endothelial growth factor( VEGF) in cerebrospinal fluid of rabbits with bacterial meningitis. Methods A total of 36 rabbits were assigned to study,which were randomly divided into meningitis model group (MOD) .dexametha-sone-treated group (DEXT) and control group( CON). CSF was sampled for determining at 6 h, 12 h and 24 h after injection of E. coli suspension. The concentration of VEGF in every CSF sample was determined quantitatively by ELISA. Results There were higher concentrations of CSF VEGF at6h,12h and 24hin M0D(( 1219 ±176) ng/L,( 1343 ±160) ng/L,(981 ±134) ng/L) than that in CON( (374 ±172) ng/L, (370 ± 169) ng/L,(367 ± 171) ng/L) (P<0.01). There was higher brain water content in M0D( (80.8 ± 0.5) % ) than that in CON( (80.0 ± 0.5) % ) (P < 0.01). There was positive correlation between the brain water content and the concentration of CSF VEGF at 24 h( r - 0.919,P < 0.01). Compared with MOD, the concentrations of CSF VEGF in DEXT at 6 h, 12 h,24 h ((941 ±147) ng/L, (1083 ± 123) ng/L, (825 ± 66) ng/L) were decreased significantly(P <0.05), the brain water content was less ((80.4 ±0.5) %) (P < 0.05). Conclusion The secretion of VEGF markedly increases in the pathological process of bacterial meningitis. VEGF contributes to the damage of blood brain barrier and the formation of brain edema. DEX can decrease the degree of brain edema by suppressing the generation of VEGF and lightening the damage of blood brain barrier.