Objective To investigate the clinical value of glypican-3 (GPC3) in the diaganosis of hepatocellular carcinoma (HCC),the contents of GPC3 in the serum and tissues of HCC patients were detected.Methods ELISA and immunohistochemical staining were applied to detect GPC3 expressing level in the serum and tissues in 79 cases with HCC,35 cases with post-hepatitis cirrhosis and 30 normal liver specimens and the resuits were compared.The influential factor of GPC3 content in the patients with HCC was analyzed by logistic regression model.Results The serum level of GPC3 in patients with HCC was (143.02±40.26) μg/L which was signifcantly higher than that in patients with post-hepatitis cirrhosis [(6.15±4.31) μg/L] and healthy controls [(4.47±3.22) μg/L] (all P ＜ 0.01).The expression levels of GPC3 was signifeantly higher in post-hepatitis cirrhosis tumor-adjacent tissue and tumor-distant tissue.The expression levels of GPC3 was significantly positively correlated with tumor presence of distant mestasis (x2 =13.182,P ＜ 0.0) and clinical stage (x2 =4.250,P ＜ 0.05),and not correlated with sex,age,tumor size and the level of AFP inserum (P ＜ 0.01).Conclusion GPC3 is specific the diagnosis of HCC.The joint diagnosis of GPC3 and AFP will improve the sensitivity of HCC.Therefore,GPC3 could as a biomarker for evaluating HCC condition and prognsis.

Objective: To observe the change of stromal cell-derived factor 1α/cysteine X cysteine receptor 4 (SDF-1α/CXCR4) signaling pathway during the process of axial stress stimulation promoting bone regeneration, and to further explore its mechanism. Methods: A total of 72 male New Zealand white rabbits were selected to prepare the single cortical bone defect in diameter of 8 mm at the proximal end of the right tibia that repaired with deproteinized cancellous bone. All models were randomly divided into 3 groups ( n=24). Group A was treated with intraperitoneally injection of PBS; Group B was treated with stress stimulation and intraperitoneally injection of PBS; Group C was treated with stress stimulation and intraperitoneally injection of AMD3100 solution. The X-ray films were taken and Lane-Sandhu scores of bone healing were scored at 2, 4, 8, and 12 weeks after operation, while specimens were harvested for HE staining, immunohistochemical staining of vascular endothelial growth factor (VEGF) and CXCR4, and Western blot (SDF-1α and CXCR4). The bone healing area was scanned by Micro-CT at 12 weeks after operation, and the volume and density of new bone were calculated. Results: X-ray film showed that the Lane-Sandhu scores of bone healing in group B were significantly higher than those in groups A and C at 4, 8, and 12 weeks after operation ( P<0.05). Micro-CT scan showed that the bone defect was repaired in group B and the pulp cavity was re-passed at 12 weeks after operation. The volume and density of new bone were higher in group B than in groups A and C ( P<0.05). HE staining showed that the new bone growth in bone defect area and the degradation of scaffolds were faster in group B than in groups A and C after 4 weeks. The immunohistochemical staining showed that the expressions of VEGF and CXCR4 in 3 groups reached the peak at 4 weeks, and group B was higher than groups A and C ( P<0.05). Western blot analysis showed that the expressions of SDF-1α and CXCR4 in group B were significantly higher than those in groups A and C at 4 and 8 weeks after operation ( P<0.05). Conclusion: Axial stress stimulation can promote the expression of SDF-1α in bone defect tissue, activate and regulate the CXCR4 signal collected by marrow mesenchymal stem cells, and accelerate bone regeneration in bone defect area.

Objective:To investigate the influence of all-trans-retinoic acid (ATRA) on the proliferation and invasion of osteosarcoma 143B cells and its possible regulatory mechanism.Methods:Different concentrations of ATRA were used to treat human osteosarcoma 143B cells, and the optimal concentration and treatment time those affected cell proliferation were selected. The MTS method, Transwell migration and invasion experiments were used to detect the changes in the proliferation, migration and invasion of 143B cells after ATRA treatment. The real-time quantitative polymerase chain reaction (qRT-PCR) and Western blot were used to detect the expression changes of miRNA-34a (miR-34a), E2F1 and Eag1 in osteosarcoma 143B cells after ATRA treatment. Then miR-34a was interfered and E2F1 was overexpressed, and the abilities of cell proliferation, invasion and migration abilities as well as the expression changes of miR-34a, E2F1 and Eag1 in 143B cells were detected.Results:The proliferation inhibition of 143B cells was most obvious when 143B cells were treated with 10 μmol/L ATRA for 72 h. The cell migration and invasion numbers when 143B cells were treated with 10 μmol/L ATRA for 72 h were lower than those in the negative control group [(73±3) cells vs. (182±5) cells, t = 21.46, P<0.01; (94±3) cells vs. (203±7) cells, t = 13.70, P<0.01]. 10 μmol/L ATRA could promote the expression of miR-34a in 143B cells and inhibit the expressions of Eag1 and E2F1 (all P<0.01). Compared with ATRA group, the ability of cell proliferation in ATRA+miR-34a interference group was restored after 72 h of treatment [cell survival rate (41.0±2.2)% vs. (25.0±3.6)%, t = 108.68, P<0.01]. Compared with ATRA group, the abilities of cell migration and invasion in ATRA+miR-34a interference group were restored [(122±14) cells vs. (64±10) cells, t = 21.06, P<0.01; (103±10) cells vs. (59±8) cells, t = 24.27, P<0.01), and the mRNA and protein expressions of Eag1 and E2F1 in cells were promoted (both P<0.01). Compared with ATRA group, the ability of cell proliferation in ATRA+E2F1 overexpression group was restored [cell survival rate (40.0±3.4)% vs. (24.0±3.1)%, t = 108.74, P<0.01]; the abilities of cell migration and invasion in ATRA+E2F1 overexpression group were restored [(78±12) cells vs. (29±8) cells, t = 13.52, P<0.01; (75±12) cells vs. (49±10) cells, t = 6.28, P<0.01], and the mRNA and protein expressions of Eag1 and E2F1 in cells were promoted (both P<0.01). Conclusion:ATRA inhibits the proliferation and invasion of osteosarcoma cells via regulating miR-34a-E2F1-Eag1 signaling pathway, and it may become one of the effective treatment drugs for osteosarcoma.

Objective To analyze changes in the percentages of circulating follicular helper T (Tfh) cells and Tfh subsets in patients with systemic lupus erythematosus (SLE) for better understanding their relationships with SLE , and to investigate effects of steroids on circulating Tfh cells .Methods Pe-ripheral blood mononuclear cells (PBMCs) from 27 patients with SLE (including 10 inactive patients and 17 active patients ) and 21 sex-and age-matched healthy donors were analyzed by flow cytometry to detect the percentage of CD4+CD45RA-CXCR5+Tfh-like cells.Disease activity and the concentration of anti-double-stranded DNA ( anti-dsDNA) antibody were evaluated by SLEDAI score ( SLE disease activity index ) and ELISA, respectively.PBMCs from healthy donors were treated with or without prednisone to evaluate its effects on circulating Tfh cells .Twelve patients with SLE were treated with high-dose steriods ( 200-500 mg/d, 2-3 d) and the percentages of circulating Tfh cells and Tfh subsets in them were analyzed before and after treatment .Results No significant difference in the percentage of circulating Tfh cells was ob-served between patients with SLE and healthy donors (P>0.05), but the percentage of Tfh17 cells in pa-tients with SLE was significantly higher than that in healthy donors (P<0.05).Compared with patients with inactive SLE and healthy donors , patients with active SLE had a lower percentage of Tfh 1 cells (P<0.05).Moreover, the percentage of Tfh1 cells was negatively correlated with SLEDAI score (r=-0.44, P<0.05). The percentage of Tfh2 cells in anti-dsDNA antibody-positive group was significantly higher than that in anti-dsDNA antibody-negative group (P<0.05).In vitro treatment of PBMCs from healthy donors with predni-sone could significantly down-regulate the percentage of circulating Tfh (P<0.01), Tfh1 (P<0.05) and Tfh2 cells (P<0.01), and up-regulate the percentage of Tfh17 cells (P<0.01).In vivo treatment of pa-tients with SLE with steriods could significantly down-regulate the percentage of circulating Tfh (P<0.01), Tfh1 (P<0.05) and Tfh2 cells (P<0.01) and up-regulate the percentage of Tfh17 cells (P<0.01).Con-clusion Abnormal distribution of Tfh subsets is correlated with SLE disease activity and anti -dsDNA anti-body .Steroids in the treatment of SLE could affect the percentage of circulating Tfh cells and the distribution of Tfh subsets .