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Zhang X.,Liaoning Medical University | Liang D.,Chinese PLA General Hospital | Liang D.,Troops of 95935 Unit PR China | Guo B.,Shenyang Orthopaedics Hospital | And 5 more authors.
Biological Trace Element Research | Year: 2013

Zinc (Zn) is an essential micronutrient and cytoprotectant involved in preventing many types of epithelial-to-mesenchymal transition (EMT)-driven fibrosis in vivo. The zinc-transporter family SLC30A (ZnT) is a pivotal factor in the regulation of Zn homeostasis. However, its function in EMT in peritoneal mesothelial cells (PMCs) remains unknown. This study explored the regulation of zinc transporters and the role they play in cell EMT, particularly in rat peritoneal mesothelial cells (RPMCs), surrounding glucose concentrations and the molecular mechanism involved. The effects of high glucose (HG) on zinc transporter gene expression were measured in RPMCs by real-time PCR. We explored ZnT7 (Slc30A7): the effect of ZnT7 over-expression and siRNA-mediated knock-down on HG-induced EMT was investigated as well as the underlying molecular mechanisms. Over-expression of ZnT7 resulted in significantly inhibited HG-induced EMT in RPMCs, while inhibition of ZnT7 expression using a considerable siRNA-mediated knock-down of RPMCs increased the levels of EMT. Furthermore, over-expression of ZnT7 is accompanied by down-regulation of TGF-β/Smad pathway, phospho-Smad3,4 expression levels. The finding suggests that the zinc-transporting system in RPMCs is influenced by the exposure to HG. The ZnT7 may account for the inhibition of HG-induced EMT in RPMCs, likely through targeting TGF-β/Smad signaling. © 2012 Springer Science+Business Media New York. Source


Zhang X.,Liaoning Medical University | Liang D.,Chinese PLA General Hospital | Guo B.,Shenyang Orthopaedics Hospital | Deng W.,Liaoning Medical University | And 4 more authors.
Cellular Signalling | Year: 2013

Zinc is an essential micronutrient and cytoprotectant involved in many types of apoptosis. The zinc transporter family SLC30A (ZnTs) is an important factor in the regulation of zinc homeostasis; however, its function in apoptosis in peritoneal mesothelial cells (PMCs) remains unknown. This study explores the regulation of zinc transporters and how they play a role in cell survival, particularly in rat peritoneal mesothelial cells (RPMCs), surrounding glucose concentrations, and the molecular mechanism involved. The messenger RNA (mRNA) transcripts were quantitatively measured by real-time polymerase chain reaction for all known nine zinc transport exporters (SLC30A1-8,. 10), as well as in primary RPMCs and the cells cultured under nonstimulated and HG-stimulated conditions. While many zinc transporters were constitutively expressed, ZnT5 mRNA and ZnT7 mRNA were strongly induced by HG. Overexpression of ZnT5 and ZnT7 respectively resulted in a decrease in the expression of caspace 3, caspace 8, BAX, and AIF and coincided with cell survival in the presence of HG. Inhibition of ZnT5 and ZnT7 expression using considerable siRNA-mediated knockdown of RPMCs was examined and, afterwards, the impact on cell apoptosis was investigated. Increased levels of apoptosis were observed after knockdown of ZnT5 and ZnT7. Furthermore, overexpression of ZnT5 and ZnT7 is accompanied by activation of PI3K/Akt pathway and inhibiting HG-induced apoptosis. This study suggests that the zinc transporting system in RPMCs is influenced by exposure to HG, particularly ZnT5 and ZnT7. This may account for the inhibition of HG-induced RPMC apoptosis and peritoneum injury, likely through targeting PI3K/Akt pathway-mediated cell survival. © 2012. Source


Liang X.,Shenyang Orthopaedics Hospital
Journal of Clinical Rehabilitative Tissue Engineering Research | Year: 2010

BACKGROUND: With follow-up prolonged, more patients received total hip replacement need to revision. Periprosthetic osteolysi, accompanied by aspetic loosening, would be aggratate, which lead to artificial joint loosening, eventually, result in joint renovation. OBJECTIVE: To observe the therapeutic effect of non-bone cement prosthesis on joint renovation following total hip replacement. METHODS: Totally 41 patients (41 hips) received renovation following total hip replacement at the Shenyang Orthopaedics Hospital from 2004 to 2009 were selected. They were renovated by Circumflex type total hip joint and press fit type non-bone cement type total hip joints. Eight cases had acetabular deficiency, 17 cases had Gustillo I-II acetabular loosening, and 8 cases had Gustillo III acetabular loosening. All these patients were received spiral mortar shaping of pure titanium directly or combined with particle bone graft. Eight patients had Gustillo ?acetabular loosening combined with bone defects, who adopted particles to plant the bone, rebuilt the hip mortar by titanium mesh, and shaping with bone cement. Non-bone cements prosthesis or lengthened prosthesis, as well as bone graft, was selected individualized. The cerclage band could be used if necessary. RESULTS AND CONCLUSION: Eight patients received titanium mesh and bone graft could do un-bearing exercise, and the remained patients could perform bearing exercise at day 3 after renovation. In the 6-66 months follow-up, no prosthetic displacement or subsidence could be found. No patients need a second renovation. Harris scores were increased from 32.6 before operation to 88.1 after operation. X-ray film showed that the bone density was increased in parts of patients. The results demonstrated that hip renovation using non-bone cement prosthesis can received an excellent short-term therapeutic effect, but the long-term effect needs to be explored. Source


Ma X.,Shenyang Orthopaedics Hospital | Ma X.,Shenyang University | Yang Y.-X.,Emporia State University | Wang Y.-F.,Shenyang University | And 3 more authors.
Journal of Dalian Medical University | Year: 2012

[Objective] To observe its effect on proliferation and invasion capacity in vitro of MG-63 cells, by application of RNAi technology to inhibit the expression of S100A4 gene in human osteosarcoma MG-63 cells. [Methods] The siRNA vector of S100A4 was constructed. According to target mRNA sequence, three RNA interference and non-specific target sequence of siRNA were designed. The cultured cells were divided into Group C (control group), group C1 (liposome transfection group), group C2 (non-specific siRNA transfection group), groups S1, S2, S3 (transfected with specific siRNA I, II, III). The situation of transfection was observed by fluorescence inverted microscope. The changes of S100A4 gene expression of the cells before and after transfection were detected by real-time PCR method. The changes of S100A4 protein expression before and after transfection were detected by Western-Blot. The specific siRNA with high transfection efficiency was selected and used as the transfection experiment group for the following tests. The following test was divided into normal cell group, negative control group and experimental group. The changes of cell proliferation were determined by MTT and flat colony-forming experiment. The changes of cell invasion and metastasis ability were determined by transwell chamber assay. [Results] Green fluorescence could be seen in cytoplasm of group C2 and group S1, S2, S3 by fluorescence inverted microscope. The expression levels of S100A4 mRNA in each specific siRNA transfection group were down regulated at various degrees. The group S3 was the most significant one. The expression levels of S100A4 protein of Western-blot detection were similar to the conclusions. MTT method and flat colony-forming experiments had shown that cell proliferation was inhibited in group S3. There was significant difference compared with the normal cells group and non-specific group (P < 0.05). Transwell chamber experiments showed that the invasive ability of the cells significantly decreased after RNA interference S100A4 gene expression, the cell number through artificial basement membrane decreased significantly, compared with the normal cell group and negative group(P <0.05). [Conclusion] S100A4 siRNA-specific vector in vitro synthesis can effectively inhibit the expression of S100A4 in osteosarcoma MG-63cells, and cell proliferation and invasive ability in vitro. Source


Ma X.,Shenyang Orthopaedics Hospital | Ma X.,Shenyang University | Wang Y.-F.,Shenyang University | An G.-F.,Shenyang University | And 2 more authors.
Journal of Dalian Medical University | Year: 2011

[Objective] To detect the expression of S100A4 gene in human osteosarcoma MG-63 and U-20S cell lines and the expressions of S100A4 and CD44V6 in osteosarcoma tissues, and then analyze its clinical significance. [Methods] The expressions of S100A4 gene in human osteosarcoma MG-63 and U-20S cell lines were detected by real-time PCR. The protein expressions of S100A4 and CD44V6 were detected by immunohistochemistry SP method, and anlyzed the relationship between both things. [Results] The result of real-time PCR showed that human osteosarcoma cell lines of MG-63 and U-20S had the expression of S100A4 gene. The positive rate of S100A4 protein expression in osteosarcoma tissues was 69.7%, and the expression located in cytoplasm. The positive rate of CD44V6 protein expression in osteosarcoma tissues was 60.6%, and the expression of CD44V6 located at cell membrane and cytoplasm near the cell membrane of osteosarcoma cells and osteochondroma cells. The expression of S100A4 was positively related to that of CD44V6 in osteosarcoma tissues (r = 0.413, P < 0.05). [Conclusion] S100A4 gene expresses in human osteosarcoma cells in vitro and in vivo. S100A4 might play a role in invasion and metastasis of osteosarcoma. Source

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