Objective:To investigate the relationship between hypercoagulable state and the activation of the NF- kappa B pathway,inflammatory/suppression cytokines in patients with osteoarthritis. Methods:56 patients with OA were divided into two groups according to random number table:XFC group ( 28 cases ) and glucosamine ( GS ) group ( 28 cases ) . Two groups were treated for 3 months. Nine healthy people are healthy control group ( NC) . Determining the expression levels of the index of the NF-κB signaling pathway (p50,p65,TAK1,IκBα) and TNF-α,IL-1,IL-10,platelet activating factor(PAF) in serum by enzyme-linked immunosorbent assay. Detected the level of the indicators and laboratory indexes related with coagulation,observed the changes between the two group, used OA symptoms integral scale, LequesneMG, SF-36 and vas to assess efficacy;and made a correlation analysis. Results: After treatment,FIB,D-D, PAF, PLT, p50, p65, TAK1, IL-1, TNF-α, hs-CRP, ESR, IgG, LequesneMG, symptom integral meter, and VAS integral were significantly higher,APTT,PT,PAF-AH,IL-10 and each dimension integral of SF-36 significantly decreased in 2 groups (P<0. 05). XFC group was better than the GS group in reducing the level of PLT,FIB,TNF-α,p65,TAK1,hs-CRP,ESR,symptom integral meter,and VAS integral and increasing PT,each dimension integral of SF-36, etc (P<0. 05,P<0. 01). The results of Pearson correlation analysis show that PLT,FIB,D-D,PAF had positive correlation with p50,p65,TAK1,IL-1,TNF-α,hs-CRP,ESR,IgG,Le-quesneMG,symptom integral meter,and VAS integral,and negative correlation with IL-10 and each dimension integral of SF-36 ( P<0. 05,P<0. 01). PT had positive correlation with IL-10,GH and PF,and negative correlation with p50,p65,TAK1,IL-1,TNF-α,hs-CRP,ESR,IgG,symptom integral meter,and VAS integral (P<0. 05,P<0. 01). Conclusion: XFC could inhibit the NF-κB signaling pathway, raise the level of IL-10, reduce the expression of IL-1, TNF-α, P50, p65, TAK1 and so on, and reduce the abnormal inflammatory immune response. So as to achieve the purpose of delaying and inhibiting the production of hypercoagulable state,reduce joint disease,relieve the symptoms of joint pain and stiffness,eventually improve the patient’s quality of life.

Objective:To observe the changes in coagulation parameters, peripheral blood cytokines, NF-kappa B signaling pathway protein, laboratory indexes in Sjogren′s syndrome ( SS ) patients, thus to explore the mechanism of hypercoagulable state. Methods:60 patients with SS and 20 healthy persons were randomly selected as the study group and the control group;Automatic coagulation analyzer was used to detect the value of coagulation parameters [ activated partial thromboplastin time ( APTT) ,prothrombin time ( PT) , plasma fibrinogen ( FIB ) , prothrombin time ( TT ) , D-dimer ( DD ) ]; ELISA method was performed to observe the expression of related cytokines(IL-1β,TNF-α,IL-10) and NF-κB signaling pathway proteins (p65,p50,IκBα); Westergren method was used to determine erythrocyte sedimentation rate ( ESR) ,and automatic biochemical analyzer to examine immune protein( IgG,IgA, IgM,GLO) and high sensitivity C-reactive protein (hs-CRP). Results: Blood coagulation parameters in 60 patients with SS were at least one abnormal for 46 cases,accounting for 76. 7% of the subjects. Among them,the abnormal rate of D-D was the highest,followed by FIB,APTT,PT,TT. Compared with the control group,D-D,FIB were significantly increased in SS patients,and TT,PT,APTT was not found obviously different. In addition,IL-1β,TNF-α,P50,P65,IκBα and inflammatory indexes like ESR,hs-CRP,IGg,GLO,ESSDAI, corneal staining score increased while the salivary flow rate,tear film break-up time and IL-10 decreased significantly( P<0. 05 or P<0. 01). Correlation analysis showed that the coagulation parameters FIB were positively correlated with the salivary flow rate,TNF-α, P50,P65,ESR,hs-CRP,while negatively correlated with IL-10,that TT was negatively correlated with TNF-α;that D-D was positively correlated with TNF-α, IL-1β, P65, ESR, hs-CRP, ESSDAI and corneal staining score, while negatively correlated with IL-10. Conclusion: SS patients generally have hypercoagulable state, and may be associated with the imbalance of cytokines, abnormal activation NF-kappa B signaling pathway,which mediates vascular endothelial cell damage,causing coagulation/fibrinolytic system dis-orders.

Objective: To investigate the clinical significance of detection of circulating tumor cells (CTCs) in peripheral blood from patients with osteosarcoma (OS) using the iFISH (immunofluorescence and fluorescence in situ hybridization) method. Methods: The live cells recovery rate of immune-magnetic beads was evaluated by live-cell fluorescent tracer technology. The expression of CD45 and CK18 on the cell surface of HOS and HepG2 cells was measured by flow cytometry. And the chromosome aneuploidy was detected by centromeric FISH probe CEP8. Subsequently, 23 OS patients were enrolled and divided into two groups, relapse or metastasis group and primary group. And the prognostic significance of CTCs numbers was analyzed. Results: The live cells recovery rate of immune-magnetic beads was higher than 90%. The flow cytometry results showed that HOS cells were double negative for the surface biomarkers of CD45 and CK18. In addition, the FISH-CEP8 signal abnormality rate were 96.5% in HOS cells. Thus, CTC was identified using the criteria as follows: the cells with CEP8-positive signal >2 accounted for more than 96.5% of the total cells, of which the cells with >3 positive signal were more than 65.0%. Among the enrolled patients, 19 patients had detectable CTCs in the peripheral blood. The CTCs numbers in the relapse or metastasis group and primary group were 2.846±1.281 and 1.400±1.506, respectively. The results showed that the CTCs in patients with recurrence or metastasis were significantly higher than those in primary patients (P=0.021). Conclusions: To our knowledge, this is the first evidence of existence of CTCs in OS patients. The CTCs numbers were positively associated with disease progression and poor prognosis. These results may provide a potential prognostic tool for monitoring metastasis and recurrence in OS patients. Trial registration Chinese Clinical Trial Registry, ChiCTR-OOC-15005925