MicroRNA (miRNA) constitutes a group of small (21-23 nucleotide) noncoding RNAs that functional as posttranscriptional gene regulators.miRNA involved in the tumor development which have been suggested to play a vital role in operating to promote or suppress tumor proliferation,invasion and metastasis.The application value of miRNA as a potential biomarker was also discussed.It needs to be further explored that miRNA can be applied as biomarkers in further medical paractice.

Objective To study the effect of alpha-melanocyte stimulating hormone (α-MSH) and its novel analogue ( STY39 ) on the production of tissue factor ( TF ) and tissue factor pathway inhibitor (TFPI) stimulated by lipopolysaccharide (LPS) in primary mouse brain microvascular endothelial cells (MBMECs). Methods Female BALB/c mice were selected,purified and primarily cultured for 5 to 7 days. Immunofluorescence assay was use to detect the Ⅷ factor related antigen and identify the MBMEC model. The MBMECs were divided into eight groups:PBS control group, LPS stimulation group, after LPS stimulation 1,2,and 3 h adding 10 -7 mol/Lα-MSH groups or STY39 group (LPS+α-MSH,LPS+STY39) ( n=4 wholes in each group) . The cell culture supernatant and cells were collected at 6 and 8 h after LPS stimulation. An enzyme-linked immunosorbent assay was used to detect the concentrations of TF and TFPI in cell supernatant. RT-PCR was used to detect the expression levels of TF mRNA. Results (1) LPS could induce MBMEC to produce TF and TFPI proteins. The level of TF in the cell culture supernatant reached the peak at 6 h,and the level of TFPI reached the peak at 8 h. (2) At 1,2,and 3 h after LPS stimulating MBMEC,10 -7mol/L α-MSH or STY39 were given. They could significantly decrease the TF protein content in the cell supernatant (P<0. 01),especially the effects of giving α-MSH or STY39 were most significant at 1 h after LPS stimulation (P<0. 05). The effect of STY39 for decreasing TF content was more significant than that of α-MSH (P<0. 05);however,α-MSH and STY39 did not have significant up-regulating effects for LPS inducing MBMEC to produce TFPI. (3) After LPS stimulation,10 -7 mol/Lα-MSH or STY39 were given at different time points. They significantly down-regulated the expression level of MBMEC TF mRNA (P<0. 01). The effect was most significant at 1 h time point (P<0. 05),but there was no significant difference in the effects betweenα-MSH and STY39. Conclusion Bothα-MSH and STY39 can suppress LPS-induced primary MBMEC to produce TF protein and express TF mRNA,and the effect of administration is better after 1 h LPS stimulation. The suppressive effect of STY39 on the production of TF protein is superior toα-MSH.

Objective To evaluate the effect of α-melanocyte stimulating hormone (α-MSH) and its novel analogue STY39 on the production of tissue factor pathway inhibitor (TFPI) in mice with endotoxemia.Methods Female BALB/c mice were randomly divided into eight groups with 9 mice in each group.Endotoxemia was reproduced by intraperitoneal injection of lipopolysaccharide (LPS,25 μg/kg) and D-galactosamine (D-Gal,100 mg/kg).The animals of the control group were given phosphate buffered solution (PBS) instead.In the experimental groups,the mice were injected intraperitoneally with 2.5 mg/kg α-MSH or STY39 at 1,2 or 3 hours following LPS injection.The orbital blood was collected at different time points,and tissues of lung,liver,and kidney were collected 8 hours after the administration of LPS.The plasma TFPI levels were determined by enzyme linked immunosorbent assay (ELISA),and the expression of TFPI mRNA in different tissues was determined with reverse transcription-polymerase chain reaction (RT-PCR).Results The plasma TFPI levels (μg/L) began to increase (11.84 ± 1.55) in the endotoxemia mice 4 hours after LPS challenge and reached the peak (23.49 ± 1.12) at 8 hours.α-MSH or STY39 treatment at 1,2 or 3 hours after LPS challenge could significantly increase the TFPI content,with the best drug effect at 1 hour after LPS challenge (the blood was collected 8 hours after LPS challenge,α-MSH group:58.79 ± 2.67 vs.28.49 ± 1.69,STY39 group:71.08 ± 2.13 vs.28.49 ± 1.69,both P＜0.01),and the effect of STY39 was better than that of α-MSH (P＜0.01).A small amount of TFPI mRNA expression was observed in each tissue of the healthy mice.After LPS challenge,TFPI mRNA expression was increased in all the tissues,especially in the lung,liver and kidney.α-MSH or STY39treatment at 1 hour after LPS challenge could significantly up-regulate the expression of TFPI mRNA in the lung and liver (A value,α-MSH in lung:51.10 ±2.89 vs.32.43 ±2.51,STY39 in lung:72.11 ±3.48 vs.32.43 ±2.51;α-MSH in liver:43.21 ± 2.12 vs.29.29 ± 2.06,STY39 in liver:66.82 ± 1.76 vs.29.29 ± 2.06,both P＜0.01).The treatment with STY39 at 1 hour after LPS challenge could significantly up-regulate the expression of TFPI mRNA in the kidney (A value:45.21 ± 1.80 vs.30.44 ± 2.23,P＜0.01),but the treatment with α-MSH had no obvious effect (A value:24.61 ± 1.98 vs.30.44 ± 2.23,P＞0.05).The enhancing effect of early administration of STY39 on TTPI mRNA expression in the lung,liver and kidney tissues of endotoxemia mice was more powerful than that of α-MSH (all P＜0.01).Conclusion The early administration of α-MSH or STY39 may up-regulate TFPI production in the mice with endotoxemia,and the effect of STY39 is superior to α-MSH.

Objective:To evaluate the effectiveness on the prevention of behavior problems of life skills education combining school-based and parent-involved approaches for third-grade students in China.Methods:This research was targeted at the population of third-grade children in two elementary schools in Qinhuangdao City,Hebei Province.Nine regular school classrooms were randomly divided into three groups:the intervention group(n=208),internal control group(n=209) and external control group(n=204).The intervention included 26-hour competence promotion for students and 5-hour parent training.The Rutter Scale by parent and teacher were used to evaluate the effects at pretest,posttest and 6-month follow-up.Results:Improvement was observed among children in the intervention group than those in the control groups.The statistical difference was significant(P

Objective To study the effects of miR-200b on proliferation and migration of sw620 colon cancer cells,and its regulation effect on E-cadherin expression.Methods The expressions of miR-200b in sw620 cells at 24 h and 72 h after pEGP-miR-200b transfection were detected by real-time PCR (RT-PCR).The change of the expression level of E-cadherin after miR-200b transfection was detected using the methods of RT-PCR and Western blot.The proliferation and migration abilities were measured by MTT and scratch test after miR-200b transfection.Results The expressions of miR-200b in sw620 cells at 24 h and 72 h after pEGP-miR-200b transfection raised significantly compared to the control group (t =11.579,P ＜ 0.01 ; t =11.579,P ＜0.01).MiR-200b transfection inhibited the proliferation abilities of sw620 cells.It is the most significant of the inhibitory effect on the third day and the inhibition rate was 55.34％.MiR-200b transfection markedly inhibited the migration abilities of sw620 cells.The two groups had significant difference in the migration distance of 24,48,72 h (t =11.579,P ＜0.01 ; t =10.419,P ＜0.01 ; t =6.955,P ＜0.01).The mRNA and protein expressions of E-cadherin gene increased significantly by transfecting miR-200b gene in sw620 cells (t =10.432,P ＜ 0.01 ; t =8.325,P ＜ 0.O1).Conclusion Up-regulated expression of miR-200b could inhibite the proliferation and migration abilities of sw620 colon cancer cells.The involved molecular mechanism is probably related to the change of E-cadherin expression.

Objective To investigate the relations of the expression of bcl-xL in superficial bladder tumor and prognosis. Method The expression of bcl-xL was detected by envision system in 80 cases of superficial bladder tumor. Results In patients with high expression of bcl-xL, the recurrence rate was higher than that of normal expression [71.4%(30/42) vs 50.0%(19/38) ,P < 0.05], and the recurrence of time as early as normal expression [(16.0 ± 1.2) months vs (36.0 ± 4.5) months](P < 0.05). Conclusion The expression of bcl-xL could effectively predict the prognosis of patients with bladder tumor, those who with high expression of-bcl-xL have bad prognosis.

Objective:In order to explore the anti inflammatory mechanisms of ? melanocyte stimulating hormone (? MSH), the effects of ? MSH on the production of NO and proinflammatory cytokines in astrocytes induced by LPS were investigated Methods:Rat brain astrocytes cultured in vitro were stimulated with LPS or given ? MSH with LPS stimulation NO produced in astrocytes was tested with Griess reagent IL 1, IL 6 and TNF ? secreted from astrocytes were examined by MTT assay The expression of macrophage migration inhibitory factor (MIF) mRNA was examined with semiquantitative RT PCR analysis Results:The production of NO, IL 1, IL 6, TNF ? and the expression of MIF mRNA were significantly increased in astrocytes stimulated with LPS If giving ? MSH with LPS stimulation, the production of NO, IL 1, IL 6, TNF ? and the expression of MIF mRNA were markedly decreased Conclusion:[WT5”,6BZ]It is suggested that the inhibitory actions of ? MSH on the production of NO and proinflammatory cytokines in astrocytes are related to the inhibitory effects of ? MSH on inflammation in central nervous system

Objective To study the expression of transcription factor special protein 1(Sp1) in colorectal cancer tissues and the relationship with the biological behavior .Methods The Sp1 mRNA expressions of 60 colon cancer tissues and their corresponding normal tissues were detected by real-time PCR ,and the level of target gene was calculated by ΔΔCT method .The relationships be-tween the expression of Sp1 mRNA and the different clinical features and pathological characters were determined .Results Com-pared with the matched normal tissues ,Sp1 mRNA was significantly up-regulated in the colon cancer tissues(P<0 .01) .Sp1 mRNA positive expression rate in colon cancer tissues had no significant different with sex ,age and tumors area(P>0 .05) ,but had signifi-cant different with histological grade ,Duke′s stages and lymph node metastasis(P<0 .05) .Conclusion Sp1 plays an important role in the process of occurrence and development in colon cancer .

Objective To study the expression of transcription factor Sp1 and CEA and the correlation between the two tran‐scription factors in colorectal cancer .Methods To detect expression Sp1 and CEA mRNA by Real‐Time PCR in 60 colon cancer tis‐sues and corresponding normal tissues and the results were compared with the clinical features and pathological characters .The re‐lationship between the expression of Sp1 mRNA and CEA mRNA in 60 colon cancer tissues was determined .Results The expres‐sion rates of Sp1 and CEA mRNA was detectable to highly expressed rates in colon cancer tissues than the matched normal tissues (P<0 .01) .There was no significant correlation between Sp1 and CEA mRNA expression in age ,sex ,tumor location(P>0 .05) . Sp1 and CEA mRNA was detectable to highly expressed in the different histological grade and Dukes stages .In addition ,a positive correlation was found between the expression of Sp1 mRNA and CEA mRNA(r=0 .706 ,P<0 .01) ,(0< r<1) .Conclusion Sp1 and CEA was detectable to highly expressed in colon cancer .Positively correlation occurred in Sp1 mRNA and CEA mRNA indica‐ted that Sp1 and CEA provide the new clues of genetic diagnosis and treatment .

Objective: To study the effects of transcription factor activator protein-2?(AP-2?)on invasive growth and estrogen receptor-?(ER-?) expression in human colon cancer SW620 cells,and to probe into the involved molecular mechanism.Methods: Plasmid pcDNA3.1(+)-AP-2? and pcDNA3.1(+) were transfected into SW620 cells by liposome-mediated transfection.The adhesion,invasion and migration abilities of SW620 cells were measured by metrical gel adhesion assay and modified Boyden chamber(Transwell assay).The gene and protein expression levels of AP-2? and ER-? in SW620 cells were examined by Real-time PCR,Western blotting and immunofluorescence cytochemistry.The interaction between AP-2? DNA and ER-? in SW620 cells was measured by electrophoretic mobility shift assay(EMSA) after AP-2? gene transfection.Results: Overexpression of AP-2? markedly reduced the adhesion,invasion and migration abilities of SW620 cells(all P