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Zhu X.-C.,University of Sichuan | Zhu X.-C.,Affiliated Hospital of Northern Sichuan Medical College | Liu Q.-S.,Affiliated Hospital of Northern Sichuan Medical College | Yang H.-M.,Northern Sichuan Medical College | Fang D.-Z.,University of Sichuan
Fudan University Journal of Medical Sciences | Year: 2014

Objective: To investigate the expression of end-binding protein 1 (EB1) in cervical tissues and its effect to the expressions of autophagic biomarkers, LC3 and p62/SQSTM1.Methods: The expression of EB1 protein in cervical carcinoma, cervical polyps and chronic cervicitis tissues was detected by mmunohistochemical method. The expression of EB1 in cervical cancer Siha cells was knocked down by siRNA, and then the expressions of autophagic biomarkers, LC3 and p62/SQSTM1 were detected by real-time fluorescent quantitative PCR and Western blot.Results: The EB1 mainly located on the cytoplasm which embraces peripheral of nucleus. The positive expression rate of EB1 in cervical cancer and cervical polyps tissues was significantly lower than those in cervical tissues of chronic inflammation (P<0.05), but the vast majority of interstitial cells were negatively expressed of EB1 in the EB1 positive expression cervical cancer tissues. The positive expression rate of EB1 in middle and low differentiation cervical squamous cell carcinoma tissues was significantly lower than that of high differentiation squamous cell carcinoma tissues (P<0.05). When the expression of EB1 was knocked down in cervical cancer Siha cells by siRNA, the expression of autophagic biomarkers, p62/SQSTM1 and LC3, were significantly increased at both mRNA level and protein level (P<0.05).Conclusions: EB1 in cervical cancer tissues whose autophagy is suppressed is lower expressed than in cervical chronic inflammation tissues whose autophagy is activated. Both p62/SQSTM1 and LC3|, the autophagic biomarkers, are dramatically increased in cervical cancer Siha cells when the EB1 expression was suppressed, which indicates EB1 plays a role in autophagy of cervical carcinoma. Source


Liu Y.-Q.,Chongqing Medical University | Luo X.-Y.,Institute of Rheumatology and Immunology | Jiang H.,Institute of Rheumatology and Immunology | Xia Y.-H.,Northern Sichuan Medical College | And 2 more authors.
Acta Anatomica Sinica | Year: 2014

Objective: To explore the expressions and implications of end-binding protein 1 (EB1) in autophagic cervical cancer Siha cells. Method Real-time quantitative PCR and Western blotting were introduced to detect the mRNA and protein expressions of EB1, LC3 and p62 in Siha cells treated with Hank's balanced salt solution (HBSS) or colchicine in vitro. Specific green fluoresent protein (GFP)-LC3 was then be detected to confirm the autophagosome formation by fluorescence microscopy with FlowCellect™ GFP-LC3 reporter autophagy assay kit. Result The mRNA and protein levels of LC3 and EB1 in cervical cancer Siha cells were increased significantly when prolong the duration of HBSS treatment (P < 0.05). EB1 mRNA was positively correlated with LC3 mRNA expression (P < 0.05). The expressions of p62 mRNA and protein reduced dramatically after HBSS treated (P < 0.05), and was negatively related with the EB1 mRNA (P < 0.05). The number of cells with GFP-LC3 and intensity of GFP-LC3 per cells were increased when starvation for 12 hours. However, the mRNA and protein levels of LC3, p62 and EB1 were not significantly changed when the cells treated with colchicine (P > 0.05), and we cannot detected the fluoresce signaling of GFP-LC3 in cells at this moment. Conclusion: The expression of EB1 in starved cells is dramatically increased, and accompanied with the increased expression of LC3 and autophagosome, decreased expression of p62. However, when microtubules with which EB1 interaction were destructed by colchicine, the above phenomenon disappears. All these results fully indicate the probably roles of EB1 in autophagy. Source


Liu Q.-S.,Institute of Rheumatology and Immunology | Liu Q.-S.,Northern Sichuan Medical College | Fan Q.-M.,Northern Sichuan Medical College | Jiang H.,Institute of Rheumatology and Immunology | And 5 more authors.
Journal of Xi'an Jiaotong University (Medical Sciences) | Year: 2010

Objective: To explore the effect of salvia miltiorrhiza on the apoptosis of fibroblast-like synoviosytes (FLSs) from patients with rheumatoid arthritis (RA). Methods: FLSs were harvested from RA patients by primary culture joint synovial fluid and through 3-5 passages. After treated with salvia miltiorrhiza for 24 h, cells were divided into high-dose (salvia miltiorrhiza's final concentration of 50 mg/mL), low-dose (salvia miltiorrhiza's final concentration of 0.39 mg/mL) and solvent group. The apoptosis of RA FLSs was detected by AnnexinV/FITC and PI with flow cytometry (FCM). Results: Compared with that in control group, cell apoptosis rate in high-dose and low-dose groups treated with salvia miltiorrhiza was significantly increased (P <. 0.05), and it was significantly greater in high-dose group than in low-dose group (P<0.05). Conclusion: Salvia miltiorrhiza can effectively induce the apoptosis of fibroblast-like synoviosytes from rheumatoid arthritis patients. Source


Liu Q.-S.,Northern Sichuan Medical College | Liu Q.-S.,Institute of Rheumatology and Immunology | Yuan G.-H.,Institute of Rheumatology and Immunology | Tang Z.,Northern Sichuan Medical College | And 11 more authors.
Journal of Xi'an Jiaotong University (Medical Sciences) | Year: 2012

Objective: To detect reticulated platelets (RPs) in patients with system lupus erythematosus (SLE) so as to explore the diagnostic and prognostic values of RPs for SLE. Methods: By using cytoanalyzer, platelets were analyzed in 80 SLE patients and 41 healthy subjects. According to the presence of thrombocytopenia, the SLE patients were divided into SLE plus thrombocytopenia group and non-thrombocytopenia group. Platelet-rich plasma was collected from all subjects through the intravenous blood. RPs were analyzed with flow cytometry. In addition, RPs were assayed in 11 SLE patients before and after medication. We counted the total number of megakaryocytes and classified them and studied the platelets in bone marrow of 21 SLE patients. Results: The initial RPs result was (6.73±6.02)% in SLE patients and (2.31±0.85)% in healthy subjects, with a significant difference (P<0.001). Among the 80 SLE patients, the RPs result was (10.99±7.54)% in the 15 patients, with thrombocytopenia, but (5.04±3.40)% in the 65 non-thrombocytopenia patients with a significant difference (P=0.001). The RPs result in 11 SLE patients significantly decreased from (8.59±7.01)% before treatment to (4.79±3.13)% after treatment (P=0.039). Bone marrow smear test showed that the number of platelets in the SLE patients with thrombocytopenia was significantly reduced compared with those non-thrombocytopenia ones. Thrombocytogenous megakaryocytes were significantly fewer in the patients with thrombocytopenia than in those without (P=0.007) although the two groups did not significantly differ in the number of immature megakaryocytes (P>0.05). Conclusion: RP has auxiliary diagnostic and prognostic values for SLE, especially in patients with thrombocytopenia. Thrombocytopenia in SLE patients may be related to the abnormal maturation of megakaryocytes in bone marrow. Source

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