Objective To evaluate the expression and clinical significance of miRNA in plasma of patients with primary biliary cirrhosis.Methods Plasma from 19 PBC patients,10 healthy volunteers and 10 viral hepatitis patients were selected from Shanghai Songjiang Hospital from december 2010 to January 2013.Among them 3 PBC patients' plasma and 3 healthy volunteers' plasma were detected by miRNA microarray for miRNA expression profile examination.Real-time PCR was used to verify the results of microarray,miRNA target gene predictior software was used to predict the target genes of differentially expressed miRNA.ROC was used to determine the clinical value of plasma miRNA.Results According to microarray,a total of 16 miRNAs were found to be differentially expressed.As revealed by qRT-PCR,the expression of miRNA-92a-3p and miRNA-4516 decreased while the expression of miRNA-572 and miRNA-575 were up-regulated in PBC group compared with healthy controls (P ＜ 0.05).In comparison with nonPBC cirrhosis group,only miRNA-92a-3p and miRNA-4516 were down-regulated (P ＜ 0.05).The area under the curve (AUC)of miRNA-92a-3p for the diagnosis and differential diagnosis of PBC were 0.92 and 0.84,respectively.while for The area under the ROC curve of miRNA-4516,the AUC for diagnosis and differential diagnosis PBC were 0.89 and 0.76,respectively.The optimal cut-off values for identifying PBC from healthy controls were defined as 1.26 ng/μl.for miRNA-92a-3p (sensitivity,92％ ;specificity,80％)and 1.16ng/ul for miRNA-4516 (sensitivity,85％ ;specificity,70％)respectively.The optimal cut-off values for identifying PBC from viral hepatitis were defined as 1.08 ng/μl.for miRNA-92a-3p (sensitivity,89％ ; specificity,81％)and 1.06 ng/μl for miRNA-4516 (sensitivity,77％ ;specificity,68％).Conclusion The results indicate that plasma from patients with PBC has a unique miRNA exprssion profile and these differentially expressed miRNA can be used as clinical biomarkers of PBC.

Objective To explore the effect of miR-24-2 on the in vitro proliferation of U-2OS cells. Methods U-2OS cells were randomly allocat-ed into 4 groups:miR-24-2 mimic group,miR-24-2 inhibitor group,negative control group,and normal control group. MicroRNAs were transfected into U-2OS cells using Lipofectamine?2000. miR-24-2 expression level and the proliferation of U-2OS cells after transfection were detected by real-time quantitative RT-PCR(RT-qPCR)and MTT proliferation assays,respectively. Results RT-qPCR results showed that miR-24-2 level was sig-nificantly higher in the miR-24-2 mimic group and lower in the miR-24-2 inhibitor group than those in the controls,indicating the successive trans-fection. MTT proliferation assay results proved that the cell viability was significantly lower in the U-2OS cells transfected with miR-24-2 mimic and higher in inhibitor groups compared to the control. Conclusion MiR-24-2 inhibits growth of the U-2OS cells,which could be a potential biomarker in the treatment of osteosarcoma.

Objective To investigate the effects of urotensin Ⅱ/urotensin Ⅱreceptor ( UⅡ/UT) system on the expression of inflammatory signal molecules p 38 mitogen-activated protein kinase ( p38 MAPK) and nuclear factor-κB ( NF-κB ) in lipopolysaccharide ( LPS )-stimulated Kupffer cells ( KCs ) . Methods Rat KCs were isolated and purified by means of in situ perfusion and density gradient centrifuga-tion.The isolated cells were randomly divided into six treatment groups including group 1:UⅡ(-) urantide (-)LPS(-), group 2:UⅡ(+)urantide(-)LPS(-), group 3: UⅡ(-)urantide(+)LPS(-), group 4:UⅡ(-)urantide(-)LPS(+), group 5:UⅡ(+) urantide(-) LPS(+) and group 6:UⅡ(-)urantide(+) LPS(+) .Western blot assay was performed to detect p 38 MAPK/p-p38 MAPK protein and NF-κB p65 sub-unit.The DNA-binding activity of NF-κB was tested by electrophoretic mobility shift assay (EMSA).Re-sults There was no significant difference with the expression of p 38 MAPK protein in KCs among the six groups (P>0.05).The expression of p65 protein and p-p38 MAPK and the DNA-binding activity of NF-κB were significantly enhanced in LPS-stimulated KCs from groups 4, 5 and 6 in comparison with those in group 1 (P<0.01).No significant differences with the levels of p65 protein and phosphor-p38 MAPK and the DNA-binding activity of NF-κB were observed between UⅡ/urantide-treated cells ( group 2 or group 3) and untreated cells (group 1) (all P>0.05), but that were decreased in group 6 than those in group 4 (all P<0.01).Conclusion UⅡ/UT system participated in the activation of p38 MAPK and NF-κB signaling pathways in LPS-stimulated primary Kupffer cells .

Objective To investigate the treatment and prevention strategies of hematomas in operation area after anterior approach surgery for cervical spondylosis.Methods A retrospective review was conducted on 12 with hematoma compression in operation area out of 785 patients managed by anterior cervical surgery from January 2007 to July 2013,including 10 males and 2 females at age ranging from 40-71 years (mean 56.8 years).Surgery method was anterior cervical corpectomy and interbody fusion using titanium mesh cage plus plate and intraoperative blood loss was 300-1 200 ml.Primary clinical manifestations were neurological dysfunction in 5 patients,dyspnea in 6,and both neurological dysfunction and dyspnea in 1.There were 10 patients with the presence of symptoms at postoperative 0.5-22 hours,1 at postoperative 73 hours,and 1 at postoperative 74 hours.All the 12 patients underwent a second anterior cervical exploration.Results There were 5 patients with epidural hematoma,6 with subcutaneous hematoma,and 1 with both hematomas.After surgical interventions,the patients presented improvement in respiratory and neurological function,with inapparent respiratory abnormality and improved neurological function at discharge.One patient was died of cardiovascular-associated disease after being discharged from hospital.The left 11 patients were followed up for mean 19.8 months (range,6-43 months),with improved Japanese Orthopedic Association (JOA) score at final follow-up.Conclusions Hematoma took place frequently in the early period,especially within 24 hours in operation area after anterior approach to cervical disorders and close attention should be paid to respiratory and limb sensation and motion functions.Early detection and early surgical interventions are the key countermeasures to avoiding the severe results.

Objecitve:To investigate effects of urotensin Ⅱ( UⅡ)/UT system on innate immune inflammatory signal pathway TLR4-IRF3 in the lipopolysaccharide (LPS)-stimulated Kupffer cells (KCs).Methods: Rat KCs were isolated and cultured.Pro-in-flammatory cytokines including IL-6,IFN-βand IFN-γwere assayed by ELISA in culture supernatant of KCs.Cell surface TLR4 were tested with flow cytometry technique.Expression of IRF3 were tested with real-time PCR and Western blot.Results: Significant increases were showed in IL-6, IFN-βand IFN-γsecretion, TLR4-expressed positive rates and IRF3 mRNA levels in KCs after stimulated by LPS,but were inhibited via urantide pretreatment.In addition,LPS induced upregulation of nuclear IRF3 protein and downregulation of cytoplasm IRF3 protein in KCs,which were blocked by urantide pretreatment.Conclusion:UⅡ/UT system mediates immune inflammatory response in part through activating TLR 4-IRF3 pathway in LPS-stimulated KCs.