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.

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.

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.

The antinociceptive effect of epidural administration of huwentoxin-I was elucidated in a tonic visceral pain rat model produced by acute colon inflammation. The nociceptive behaviors were induced by perendoscopically injecting dilute formalin (50 μl) into the depth of the colonic wall in rats. Both ω-conotoxinMVIIA and morphine hydrochloride were given epidurally as positive control while saline as negative control.Similar to ω-conotoxin-MVIIA and hydrochloride morphine, the epidural administration of HWTX-Ⅰ significantly reduced the nociceptive responses in a dose-dependent manner in tonic visceral pain rat model ( P ＜ 0.05). The suppression effects of both huwentoxin- Ⅰ and ω-conotoxin-MVIIA at 20 μg/kg were kept steady compared with the saline group and reached their maximum effects at the doses of 50 ～ 75 μg/kg within 1 hour when the nociception had been observed. It was also found that at the same doses, huwentoxin- Ⅰ was less effective in antinociception than ω-conotoxin-MVIIA. However, ω-conotoxin-MVIIA, but not huwentoxinⅠ , caused an obvious motor dysfunction at these doses. The action of morphine hydrochloride was initiated faster, but lasted for a shorter time than that of huwentoxin- Ⅰ and ω-conotoxin-MVIIA. Thus, huwentoxinⅠ , a potent blocker of neuronal N-type voltage-sensitive calcium channels, induced a remarkable dosedependent restrain effect similar to ω-conotoxin-MVIIA and morphine on the tonic visceral pain produced by colonic wall injection of formalin in conscious rats.

Objective To evaluate the efficacy of epidural anesthesia combined with different doses ropivacaine and sufentanil for stepwise labor analgesia in latent phase.Methods Two hundred and ten ASA Ⅰ or Ⅱ primiparas with a singleton and vertex presentation at full term in our hospital from February 201 5 to April 201 5 were randomized into seven groups (n =30 each):0.125% ropiva-caine with 0.5 μg/ml sufentanil (group 1);0.075% ropivacaine with 0.5 μg/ml sufentanil (cervical dilatation < 3 cm),0.125% ropivacaine with 0.5 μg/ml sufentanil (cervical dilatation ≥ 3 cm) (group 2);0.1% ropivacaine with 0.5 μg/ml sufentanil (cervical dilatation < 3 cm),0.125% ropiv-acaine with 0.5 μg/ml sufentanil (cervical dilatation≥3 cm)(group3);0.1 5% ropivacaine with 0.5μg/ml sufentanil (group 4);0.075% ropivacaine with 0.5 μg/ml sufentanil (cervical dilatation < 3 cm),0.1 5% ropivacaine with 0.5 μg/ml sufentanil (cervical dilatation≥ 3 cm)(group 5 );0.1%ropivacaine with 0.5 μg/ml sufentanil (cervical dilatation<3 cm),0.1 5% ropivacaine with 0.5 μg/ml sufentanil (cervical dilatation≥3 cm)(group 6);0.125% ropivacaine with 0.5 μg/ml sufentanil (cervical dilatation<3 cm),0.1 5% ropivacaine with 0.5 μg/ml sufentanil (cervical dilatation≥3 cm) (group 7).The intensity of pain was assessed by visual analog scale (VAS).Meanwhile,1abor process,postpartum hemorrhage,Bromage score,postpartum adverse reactions and Apgar score of the neonates were also observed.Results No significant difference was found in VAS score after epi-
dural block between groups at each time.The latent period of group 2 and 3 were shorter than that in group 1 (P <0.05)and that of group 5 and 6 were shorter than that in group 4 (P <0.05);the ac-tive phase of group 4 were longer than that in group 1 (P <0.05 ).The postpartum hemorrhage of group 2 and 3 were less than that in group 1 (P <0.05),the postpartum hemorrhage of group 5,6 and 7 were more than that in group 2 (P <0.05)and group 3 (P <0.05).The motor nerve block of group 2 and 3 were slightly less than that in group 1 (P <0.05)and the motor nerve block of group 5,6 and 7 were slightly less than that in group 4 (P <0.05).There was no difference of the postpar-tum adverse reactions of maternal and Apgar score in the neonates.Conclusion The dosage of 0.075% or 0.1% ropivacaine with 0.5 μg/ml sufentanil (cervical dilatation < 3 cm),0.125% ropiv-acaine with 0.5 μg/ml sufentanil (cervical dilatation ≥ 3 cm),while producing the exact analgesic effect,hardly interferes with the 1abor process,the amount of postpartum hemorrhage and the lower limb activity,thus they have no significant effect on the safety of the maternal and the infant.

OBJECTIVE ToinvestigatethepossibleprotectiveeffectofTripterygiumwilfordiipolyg-lycoside (TWP ) on experi mental diabetic nephropathy (D N ) rats and its possible mechanis m. METHODS Thediabeticmodelwasinducedbyasingleintraperitonealinjectionofstreptozotocin (STZ)65 mg·kg -1 .Three weeks after modeling,TWP 4.5,9.0 and 1 8.0 mg·kg -1 was ig given to rats,once daily,for 8 consecutive weeks.During the experiment,the changes of body mass,hair, mental health of rats were observed.Two days before the end of the experi ment,the rats were placed into metabolic cages to collect 24 h urine in order to detect 24 h urinary albu min excretion rate (UAER). The rats were given TWP for 8 weeks and anesthetized with 1 0%chloral hydrate.The blood was collect-ed fro m the heart and centrifuged,seru m creatinine and urine creatinine were measured,and creatinine clearance (Clcr)was calculated.Blood urea nitrogen (BUN)and the serum catalase (CAT)activity were tested by optical method while the level of seru m superoxide oxygen anion(O2÷)was tested by col-orimetry.The level of malondialdehyde(MDA)was determined by thiobarbituric acid condensation,and glutathione peroxidase (GSH-Px)activity was tested by colorimetry.The right kidney was HE stained to observepathologicalchanges.RESULTS Comparedwithnormalcontrolgroup,theratsinmodel control group developed polydipsia,polyuria,polyphagia,body mass loss,unresponsiveness,brown hair,pale tail and apathy clammy.Besides,blood glucose,BUN and 24 h UAER were significantly higher (P<0.01 ),but Clcr was lower (P<0.01 ).The activity of serum CAT and GSH-Px in renal tissue was significantly lower(P<0.01 ),while the level of serum O2÷ and MDA in the renal tissue was significantly higher(P<0.01 ).Compared with model control group,TWP 9.0 and 18.0 mg·kg -1 could improve the general condition of rats.BUN and 24 h UAER were obviously reduced(P<0.01 ),Clcr and serum CAT were increased obviously(P<0.01 ),the level of MDA and O2÷ were reduced obviously(P<0.01 ),and GSH-Px level was increased(P<0.01 ).TWP 9.0 and 18.0 mg·kg -1 could significantly im-prove the renal histopathological changes of rats.TWP 4.5 mg·kg -1 had no significant effect on the aboveindicators.CONCLUSION TWPhasprotectiveeffectontherenalfunctionofexperimentalDN rats.The mechanis m may be related to inhibition of the oxidative stress and enhance ment of the body antioxidant capacity.

Objective: To investigate the gene frequency of HLA-B* 5801 in the population of Chinese Minnan region.Methods:In this study,we enrolled 178 patients requiring allopurinol therapy( including 40 patients with gout,89 patients with hyperuricemia and 49 patients with gouty arthritis) and 100 healthy people.We isolated genomic DNA from their blood and screened for HLA-B*5801 with both PCR and gene sequencing.Results:We found 22%patients and 16%healthy people with HLA-B*5801.The frequencies of HLA-B*5801 in patients and healthy people are 0.13 and 0.09,respectively.The results from PCR and gene sequencing were consistent.Conclusion:The frequency of HLA-B*5801 in the population of Chinese Minnan region is relatively high.Therefore,it is necessary to screen for HLA-B*5801 in allopurinol users before taking the medicine.

Objective To investigate the genotype and epidemiology of plasmid‐mediated AmpC β‐lactamases produced by the clinical strains of Escherichia coli and Klebsiella pneumoniae .Methods A total of 176 clinical nonrepetitive cefoxitin non‐sensitivity isolates of Escherichia coli and Klebsiella pneumoniae was collected from July 2011 to August 2012 .Polymerase chain reaction (PCR) for AmpC enzyme gene amplification and DNA sequencing were carried out for genotype of AmpC beta‐lactamases .Results The results of PCR showed that the positive rate of ampC of the 176 strains of Escherichia coli and Klebsiella pneumoniae AmpC was 18 .2% ,mainly DHA type ,counting for 59 .4% ,CIT counting for 37 .5% ,EBC counting for 3 .1% .The positive rate of ampC of Escherichia coli was 11 .4% ,mainly CIT type ,counting for 77 .8% ,the positive rates of DHA type and EBC type both were 11 .1% .The positive rate of ampC of Klebsiella pneumoniae were 23 .7% ,mainly DHA type ,counting for 78 .3% ,CIT type count‐ing for 21 .7% .The results of DNA sequencing showed that there were 18 strains DHA‐1 type and 1 strain ampC gene type of Morganella morganii in DHA type strains ,the concordance rate was 97 .0% ,10 CIT type strains was CMY‐2 type ,1 strain was CMY‐42 ,one strain was CMY‐4 type ,EBC type was ampC gene type of Enterobacter cloacae ,the concordance rate was 99 .0% .A total of 32 strains of gene sequencing were registered as KJ127248 - KJ127279 in GenBank .Conclusion The main genotypes of plasmid‐mediated ampC enzyme produced by Escherichia coli and Klebsiella pneumoniae were CMY‐2 and DHA‐1 respectively .

Objective To conduct the epidemiological investigation and analysis of cerebral palsy in Xinxiang of Henan Province and to investigate its risk factors in order to provid a basis for further study of etiology and prevention of cerebral palsy information.Methods Cluster sampling survey was carried out among children aged 1-6 years in XinXiang,Henan Province,and the data were analyzed by using SPSS 13.0 statistical analysis software.Results The morbidity of infantile cerebral palsy in Xinxiang of Henan Province was 2.82‰.The prevalence distribution in all age groups was 2.46 ‰-3.11‰(x2 =0.374,P =0.996),and the prevalence rate in male and female was significantly different(x2 =0.139,P =0.709) ; the sex ratio was 1.09 ∶ 1.00.Prevalence rate was slightly lower in urban areas than in rural areas (x2 =0.526,P =0.769).But no significant differences were observed in all of the data above.The incidence of cerebral palsy of children whose mothers did not established perinatal care manual and guidance during pregnancy was 5.86 times of the children whose mothers established perinatal care manual and guidance (x2 =116.806,P =0.000) ;the incidence of cerebral palsy in children whose mothers did not receive regular prenatal care during pregnancy was 5.37 times of the children whose mothers receive regular prenatal care during pregnancy (x2 =43.904,P =0.000);the incidence of cerebral palsy in children who had no neonatal follow-up after birth was 8.55times of the children with neonatal follow-up after birth (x2 =68.987,P =0.000).The incidence of cerebral palsy in children whose developmental disorders were not timely diagnosed and treated medically was 5.39 times the children whose developmental disorders were timely diagnosed and treated (x2 =56.003,P =0.000).The significant differences were observed in all of the data above.In the classification of cerebral palsy,the spastic type was the most (42.1％) ;followed by the dyskinetic (24.6％) ; the mixed (18.8％) ; and the ataxia(14.5％).Conclusions The survey results can reflect current prevalence of infantile cerebral palsy in children aged 1-6 years in XinXiang,and can be served as a basis for further prevention and treatment of cerebral palsy information.

Objective To investigate the effect of high mobility group box‐1(HMGB1) on the migration of vascular smooth cells (VSMCs) and the role of TLR4‐dependent PI3K/Akt pathway in the process .Methods Primary VSMCs were isolated from the thoracic aorta of male SD rats and cultured in vitro .Control group ,TLR4 siRNA transfected group ,control siRNA transfected group and PI3k inhibitor (LY294002) intervention group were stimulated by HMGB1 (0 .1-1 000 .0 ng/mL) .Expression of TLR4 mRNA was detected by RT‐PCR ,protein expression of TLR4 ,Akt ,pAkt ,PI3K were detected by Western blot .Activity of the im‐munoprecipitated PI3K enzyme was assessed in a competitive ELISA .The migration and cell viability of every groups were ob‐served .Results HMGB1 (0 .1 -1 000 .0 ng/mL) stimulated VSMCs migration in a dose‐dependent manner and incubation of VSMCs with 100 ng/mL caused a rapid migration (P< 0 .05) .At the concentrations used ,HMGB1 did not cause any cytotoxic effects (P<0 .05) .Migration of VSMCs toward HMGB1 was significantly inhibited by silencing of TLR4 (P<0 .05) .Pretreated cells with TLR4 siRNA or the PI3K inhibitor LY294002 could markedly block PI3K/Akt pathway activation and VSMCs migration mediated by HMGB1 (both P<0 .05) .Conclusion HMGB1 stimulated VSMCs migration in a dose‐dependent manner and TLR4‐dependent PI3K/Akt signaling pathway played an important role in the migration of VSMCs mediated by HMGB1 .This research indicates that TLR4‐dependent TLR4/PI3K/Akt signaling pathway could be the target in the treatment of obstructive cardiovascu‐lar disease .