Objective To explore the function of olfactomedin domain family protein 3(OLFM3)in neuroblastoma(NB).Methods The relationship between the expression of OLFM3 mRNA and v-myc avian myelocytomatosis viral oncogene neuroblastoma derived homolog(MYCN)amplification status and the prognosis of patients in NB clin-ical samples were clarified by using R2 Genomics Analysis and Visualization Platform.Depmap database was used to examine the expression level of OLFM3 in different tumors cell lines and to identify the correlation between OLFM3 expression and MYCN amplification status in various NB cell lines.RT-qPCR and Western blot were used to detect the knockdown level of OLFM3.Cell proliferation was monitored using crystal violet staining and real?time cellular analysis.The colony formation ability of NB cells was assessed using colony?forming unit assay.Results Analysis of R2 database revealed higher level of OLFM3 expression in MYCN?amplified NB clinical samples(P<0.001).Patients with high OLFM3 expression showed a significantly lower overall survival probability compared to those with low OLFM3 expression(P<0.05).Analysis with Depmap database revealed that the expres?sion level of OLFM3 was higher in NB than that in other kind of tumor.The expression level of OLFM3 was signifi?cantly higher in the MYCN?amplified cell lines than in the MYCN?non?amplified cell lines(P<0.01).In MYCN?am?plified NB cells,knockdown of OLFM3 inhibited cells proliferation(P<0.001)and colony formation(P<0.001),but there was no noticeable changes observed in MYCN?non?amplified cells.Conclusions OLFM3 specifically pro?motes the proliferation of MYCN?amplified NB cells,but has a less effect on MYCN?non?amplified cells,indicating it is a potential biomarker for high?risk MYCN?amplified NB.

Objective To explore the effect of open reading frame 66(C12ORF66)located at chromosome 12 on the viability of MYCN amplified NB cell lines.Methods DDatasets GSE16476 and GSE49710 in R2 database were analyzed for expression level of C12ORF66 in MYCN amplified and MYCN non-amplified NB cells and its potential correlation with the prognosis of pediatric patients.C12ORF66 mRNA expression level in normal tissue immortalized cell lines,MYCN amplified and MYCN non-amplified cell lines were detected by RT-qRCR.Transient or stable knockdown of C12ORF66 cell lines were constructed to compare the difference in real time cellular analysis(RTCA),colony formation,Ki67 positive cells between the control group and the C12ORF66 knockdown group.Results By analyzing R2 datasets,C12ORF66 level in MYCN amplified samples was significantly higher than that in MYCN non-amplified samples,and the expression of C12ORF66 was negatively correlated with the prognosis of pediatric patients(P＜0.05).C12ORF66 highly expressed in MYCN-amplified BE(2)-C and SK-N-BE(2)cell lines than in MYCN non-amplified CHLA-255 and SH-SY5Y cell lines(P＜0.001).Transient or stable knockdown of C12ORF66 resulted in significant slow down of proliferation of MYCN amplified NB cells(P＜0.001),the colony formation ability was significantly reduced(P＜0.001),and the proportion of Ki67 positive cells was significantly decreased(P＜0.05).Conclusions C12ORF66 was highly expressed in MYCN amplified clinical NB samples and cell lines which is believed to be correlated with poor prognosis of pediatric patients.C12ORF66 knockdown signifi-cantly inhibits cell viability of NB cells.

Objective:To investigate the function and underlying mechanism of cysteine and glycine-rich protein 2(CSRP2)in neuroblastoma(NB).Methods:The correlation between the expression level of CSRP2 mRNA and the prognosis of NB children in NB clinical samples was analyzed in R2 Genomics Analysis and Visualization Platform.The small interfering RNA(siRNA)targeting CSRP2 or CSRP2 plasmid were transfected to NB cell lines SK-N-BE(2)and SH-SY5Y.Cell proliferation was observed by crystal violet staining and real-time cellular analysis.The ability of colony formation of NB cells was ob-served by colony-forming unit assay.Immunofluorescence assay was used to detect the expression of the proliferation marker Ki-67.Flow cytometry analysis for cell cycle proportion was used with cells stained by propidium iodide(PI).Annexin V/7AAD was used to stain cells and analyze the percentage of cell apoptosis.The ability of cell migration was determined by cell wound-healing assay.The level of protein and mRNA expression of CSRP2 in NB primary tumor and NB cell lines were detected by Western blot and quantitative real-time PCR(RT-qPCR).Results:By analyzing the NB clinical sample databases,it was found that the expression levels of CSRP2 in high-risk NB with 3/4 stages in international neuroblas-toma staging system(INSS)were significantly higher than that in low-risk NB with 1/2 INSS stages.The NB patients with high expression levels of CSRP2 were shown lower overall survival rate than those with low expression levels of CSRP2.We detected the protein levels of CSRP2 in the NB samples by Western blot,and found that the protein level of CSRP2 in 3/4 INSS stages was significantly higher than that in 1/2 INSS stages.Knockdown of CSRP2 inhibited cell viability and proliferation of NB cells.Overexpression of CSRP2 increased the proliferation of NB cells.Flow cytometry showed that the proportion of sub-G1,G0/G1 and S phase cells and Annexin V positive cells were increased after CSRP2 deficiency.In the cell wound-healing assay,the healing rate of NB cells was significantly attenuated after knockdown of CSRP2.Further mechanism studies showed that the proportion of the proliferation marker Ki-67 and the phospho-rylation levels of extracellular signal-regulated kinases 1/2(ERK1/2)were significantly decreased after CSRP2 knockdown.Conclusion:CSRP2 is highly expressed in high-risk NB with 3/4 INSS stages,and the expression levels of CSRP2 are negatively correlated with the overall survival of NB patients.CSRP2 significantly increased the proliferation and cell migration of NB cells and inhibited cell apoptosis via the activation of ERK1/2.All these results indicate that CSRP2 promotes the progression of NB by activating ERK1/2,and this study will provide a potential target for high-risk NB therapy.

Objective : To explore the correlation between air pollution and outpatient visits,and to provide evidence for health risk assessment of air pollutants and intervention. Methods : The data of air pollutants and outpatient visits in 2016 in Zhoushan Hospital were collected,and the outpatient volume on the days when the concentration of air pollutants reached and exceeded the standard were compared. Spearman rank correlation analysis was used to explore the relationship between outpatient volume and the concentration of air pollutants. Results : In 2016,the median（inter-quartile range）of daily outpatient volume in Zhoushan Hospital was 3 304（1 638）person-times. O3,PM2.5 and PM10 were the primary air pollutants in Zhoushan in 2016. The average daily outpatient volume of internal medicine,circulatory system,other diseases and all when the air was polluted at light level or above were higher than those when the air quality was good（P< 0.05）. CO concentration was positively correlated with the average daily outpatient volume of respiratory system and circulatory system（P< 0.05）,was negatively correlated with the average daily outpatient volume of all（P< 0.05）. O3-8 h concentration was positively correlated with the average daily outpatient volume of internal medicine,other diseases and all（P< 0.05）,and was negatively correlated with the average daily outpatient volume of respiratory system and pediatrics（P< 0.05）. SO2 concentration was negatively correlated with the average daily outpatient volume of respiratory system,skin and subcutaneous tissue（P< 0.05）. The concentration of NO2,PM10 and PM2.5 were positively correlated with the average daily outpatient volume of respiratory system and pediatrics（P< 0.05）. Conclusion The main air pollutants in Zhoushan were O3,PM2.5 and PM10. When they exceed the limits,the outpatient volume would increase.

Objective To investigate the roles of CD3+,CD4+,CD8+T lymphocytes and CD19+B lymphocytes on the pathogenesis of acute respiratory distress syndrome(ARDS).Methods According to Berlin definition Of ARDS in 2012,34 patients with ARDS admitted in the Department of ICU of Central Hospital of Baoji from January,2016 to January,2017 were enrolled in this study as study group(ARDS group).At the same time,22 healthy subjects were recruited as control group.Clinical data of ARDS patients were collected,and the survivors were followed up.The ARDS patients were divided into moderate group(n=20) and severe group (n=14) according to clinical settings on the first day after diagnosis of ARDS and Berlin Definition of ARDS in 2012,and at the same time they were also dividedinto two groups according to the outcome followed up for 28 days:non-survival group(n=14) and survival group(n =20).Sample of 3 mL peripheral venous blood of ARDS patients was collected on an empty stomach in the early morning on the first day after diagnosis of ARDS and the blood samples of healthy subjects were also collected on the first day to measure the level of CD3+,CD4+,CD8+T cells and CD19+ B cell in peripheral venous blood by flow cytometry.Comparison of CD3+,CD4+,CD8+ T cells and CD 19+ B cell numbers were carried out between ARDS group and control group on the first day after diagnosis of ARDS,and between moderate group and severe group as well as between survival group and nonsurvival group.The risk factors associated with ARDS were analyzed using logistic regression analysis.Results On the first day after diagnosis of ARDS,there were significant differences in serum Lac and pre-albumin between survival group and non-survival group(P＜0.05).The numbers of CD3+,CD4+T cells and CD19+B cell of peripheral venous serum in ARDS group were significantly lower than those in control group(P＜0.05),while there was no significant difference in CD8+ T cell number between ARDS group and control group (P＞0.05).There were statistically significant differences in numbers of CD3+,CD4+,CD8+T cells and CD19+B cell between moderate group and severe group and as well as between survival group and non-survival group(P＜0.05).Logistic regression analysis showed that CD19+B cell (OR=0.614,95％CI:0.416-0.907,P=0.014) level on the first day after diagnosis of ARDS was related with the risk of prognosis of ARDS.The ROC of CD19+B cell had area under curve(AUC) of 0.907,and the cut-off value of CD19+B cell in the survival followed up for 28 day's was 12.59％.Conclusions CD3+,CD4+,CD8+T cells and CD19+B cell level of peripheral venous serum in ARDS patients can be helpful for the assess of ARDS severity of patients in the early stage,and for prognosis judgment,especially CD 19+B cell is more remarkable.

Objective To study and analyze the clinical effect of valsartan on chronic pulmonary heart disease. Methods 100 cases of heart failure patients with pulmonary heart disease treated in our hospital from January 2015 to October 2016 were selected and randomly divided into the control group and the experimental group,50 cases in each group.Two groups of patients were given the basic treatment of diuretics,low salt diet, digitalis and nitrates,so as to ensure that the patients had enough rest.Patients in the control group were treated with benazepril,and the patients in the experimental group were treated with valsartan.The clinical indexes of the experimental group and the control group were compared and analyzed. Results After corresponding treatment,the number of effective treatment was 48 cases.In the control group,the total effective number was 41 cases. The effective rate of the control group (82.0％) was significantly lower than the experimental group(96.0％) (P<0.05).The incidence of adverse reactions in the experimental group was 8.0％, and the fatality rate was 10.0％. The incidence of adverse reactions in the control group was 10.0％, and the fatality rate was 12.0％.There was no significant difference between two groups.After treatment,the indexes of LVEDD and LVESD in the experimental group were better than those in the control group,with statistical significance(P<0.05).Conclusion Valsartan was effective in the treatment of chronic pulmonary heart disease with heart failure.It can improve the treatment efficiency and security in a certain extent,and has the significance of further popularization and application.

OBJECTIVE: To explore the relationship between the expression of interferon-induced protein with tetratricopetide repeats 1 (IFIT1) and liver cell apoptosis in the acute stress period after severe burns.
 METHODS: A total of 25 C57/129 adult mice were randomly divided into the normal control group (0 h) and the groups at 1, 6, 12 or 24 after severe burns (n=5 per group). A model with third degree (20% of the total body surface area) burn injury was established and then liver tissues were taken. IFIT1 expression was examined by Western blot. The expression of caspase-3 and -8 was measured by immunohistochemistry. Liver cell apoptosis was detected by terminal deoxynucleotidyl transferase mediated nick end labeling (TUNEL).
 RESULTS: After burns, IFIT1 expression was increased at 1 h, which reached the highest level at 
6 h followed by a decrease at 12 h, which reached minimum level at 24 h. The differences between groups were significant (P<0.01). The caspase-3 and -8 levels significantly increased after burns in a time-dependent manner (P<0.01). Although at 0 h and 1 h there was no significant increase in liver cell apoptosis, the increase reached significance from 6 h to 24 h (P<0.01).
 CONCLUSION The increase in IFIT1 expression after severe burns promotes liver cell apoptosis.
Animals ; Apoptosis ; Blotting, Western ; Burns ; metabolism ; Carrier Proteins ; metabolism ; Caspase 3 ; metabolism ; Caspase 8 ; metabolism ; Hepatocytes ; cytology ; In Situ Nick-End Labeling ; Liver ; cytology ; Mice ; Mice, 129 Strain ; Mice, Inbred C57BL ; RNA-Binding Proteins

Objective In order to discuss the contribution and significance of lowering increased serum uric acid levels in cardio-vascular disease prevention and control .Methods All 100 hyperuricemia in senile patients with hypertension and diabetes mellitus were received the standard drug treatments for bloodpressure ,blood sugar management .According to the patients′will divided them into uric acid intervention group(study group) and control group .The study group were given low purine diet and benzbromarone tablet (50 mg/day ,course of 1 year) ,then compare the changes between the 2 group and within each group after 3months ,6months and 1 years in the metabolism indexes before and after intervention (serum uric acid ,glycosylated hemoglobin ,fasting blood glu-cose ,2-hours postprandial glucose ,and the changes of dynamic blood pressure ) ,and follow-up the occurrence of cases in primary end point events (all-cause death ,total cardiovascular death) .Results (1)In study group ,3months ,6months and 1 years after in-tervention ,the serum uric acid lever was significantly lower than that before intervention and the control group ,P<0 .01 .(2)The study group patients′dynamic blood pressure was significantly lower than that before intervention and the control group after 6 ,12 months ,meanwhile the study group patients′success rate of dynamic blood pressure level is higher than themselves before the in-tervention after 6 ,12 months ,P<0 .05 .(3)The 2 groups of patients′glycosylated hemoglobin ,fasting plasma glucose ,2-hours post-prandial glucose ,control rate of blood glucose at each testing point before and after the intervention had no difference .(4)The oc-currence of cases between the 2 groups for all-cause death and total cardiovascular death had no difference .Conclusion (1)Reduc-ing the mortality and disability rate of hyperuricemia in elderly patients with hypertension and diabetes mellitus ,the most critical measures is still a reasonable standard blood pressure and blood sugar management .Based on this therapy ,effectively reducing the increased serum level of uric acid can improve the control rate of blood pressure .

OBJECTIVE: To observe the influence of ropivacaine on the proliferation and migration of rat bone marrow mesenchymal stem cells (BMSCs) and provide basis for the clinical application of BMSCs. METHODS: Rat BMSCs were isolated and cultured by adherence method. Surface markers of BMSCs were examined by flow cytometry. Multipotent differentiation of BMSCs was detected by induced adipogenesis, osteogenesis and muscular differentiation. Proliferation of BMSCs was examined by CCK-8 and Brdu incorporation after ropivacaine treatment at different concentrations. Migration of BMSCs was tested by cell scratch assay and Millicell experiment. RESULTS: Cultured cells had representative appearance and surface markers of BMSC, and they had potential multiple differentiation. Ropivacaine treatment at 50 and 100 μmol/L significantly reduced the proliferation rate of BMSCs and Brdu incorporation rate. There was significant difference compared with the control group (P<0.05). Cellular scratch assay and migration experiment indicated that ropivacaine significantly reduced the migration of BMSCs. There was significant difference compared with the control group (P<0.05). All these mentioned effects of ropivacaine on BMSCs were dose-dependent. There was significant difference between groups (P<0.05). CONCLUSION Ropivacaine can significantly reduce the proliferation and migration of rat BMSCs, suggesting that the influence of local anesthetics on BMSCs has to be taken into account when BMSCs are used in clinical practice.
Amides ; pharmacology ; Animals ; Bone Marrow Cells ; Cell Differentiation ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Flow Cytometry ; Mesenchymal Stem Cells ; cytology ; drug effects ; Rats ; Ropivacaine

Objective To compare the efficiency and safety of aspirin-dipyridamole and warfarin in the prevention of thromboembolism in patients with nonvalvular atrial fibrillation(NVAF)and high risk factors.Methods One hundred and forty NVAF patients with high risk factors were randomly divided into two groups.Warfarin group[78 cases international normalized ratio(INR)2.0-3.0,for the patients older than 75 years,INR ranging from 1.6 to 2.5]and combination group(62 cases received aspirin 160 mg once every day plus dipyridamole 160 mg 3 times every day).The incidence of death,thromboembolism(including stroke and peripheral arteries embolism)and hemorrhage events were observed.Results Followed-up 12-28 months.In warfarin group,3 cases lost,2 cages had stroke,2 cases suffered from serious bleeding events,6 cases had minor bleeding events.In combination group,2 cases lost,6 cases had stroke,and 2 cases suffered from peripheral arteries embolism events,3 cases had minor bleeding events,but no serious bleeding events occurred.The incidence of thromboembolism in warfarin group wag,lower than that in combination group[2.7%(2/75)vs 13.3%(8/60),P<0.05].There was no significant difference of the bleeding rate between the two groups[10.7%(8/75)vs 5.0%(3/60),P>0.05].Conclusions Warfarin anticoagulative therapy is more effective than aspirin and dipyridamole antiplatelet dual therapy for the prevention of thromboembolism events in patients with NVAF and high risk factors.The major bleeding events in warfarin group occurs in patients with INR>3.0,so under intensive monitoring(INR 2.0-3.0),warfarin therapy is effective and safety.