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He Z.,Shanghai Fengxian Central Hospital | Hu B.,Shanghai JiaoTong University | Tang L.,Shanghai JiaoTong University | Zheng S.,Shanghai JiaoTong University | And 4 more authors.
Journal of Cancer Research and Therapeutics | Year: 2015

Doxorubicin (Adriamycin, ADM) is an antimitotic drug used in the treatment of a wide range of malignant tumors, including acute leukemia, lymphoma, osteosarcoma, breast cancer, and lung cancer. Multidrug resistance-associated proteins (MRPs) are members of a superfamily of ATP-binding cassette (ABC) transporters, which can transport various molecules across extra- and intra-cellular membranes. The aim of this study was to investigate whether there was a correlation between MRP4 and primary ADM resistance in osteosarcoma cells. In this paper, we chose the human osteosarcoma cell line MG63, ADM resistant cell line MG63/DOX, and the patient's primary cell GSF-0686. We checked the ADM sensitivity and cytotoxicity of all the three cells by cell proliferation assay. The intracellular drug concentrations were measured by using LC-MS/MS. We also examined MRP4 gene expression by RT-PCR and Western Blot. We found that the intracellular ADM concentration of the parent osteosarcoma cell line MG63 was higher than the ADM resistant osteosarcoma MG63/DOX cell line or the GSF-0686 cell after ADM treatment (P < 0.05). In addition, MRP4 mRNA and protein levels in ADM resistant osteosarcoma cells were higher than in MG63 cell (P < 0.05). Taking together, this work suggests that overexpression of MRP4 may confer ADM resistance in osteosarcoma cells. Source


Zhang B.,Shandong University | Wang A.,Shanghai Fengxian Central Hospital | Xia C.,Shanghai Fengxian Central Hospital | Lin Q.,Shanghai Fengxian Central Hospital | Chen C.,Shandong University
Molecular Medicine Reports | Year: 2015

Alzheimer's disease (AD) is a progressive neurodegenerative disorder and is the most common form of dementia among the aging population. Although the incidence of the disease continues to increase, no cure has been developed. Effective treatment is restricted not only due to the lack of curative medicine, but also due to limited understanding of the underlying mechanisms and the difficulties in accurately diagnosing AD in its earliest stages prior to clinical symptoms. Micro (mi) RNAs (miR) have gained increasing attention in the investigation of neurodegenerative diseases. Previous reports have demonstrated that deregulation of miR-146a-5p is associated with the pathogenesis of human AD. In the present study, the coding region of primary (pri)-miR-146a in patients with AD was scanned and the rare C allele of rs2910164 was found to be associated with AD. Using reverse transcription quantitative polymerase chain reaction, it was demonstrated that site variation reduced the expression of mature miR-146a-5p. Notably, a reduction in the expression of miR-146a-5p led to less efficient inhibition of target genes, including Toll-like receptor (TLR)2, which is important in the pathogenesis of AD. Biological function investigations in RAW264.7 cells indicated that, compared with the G allele, the rare C allele upregulated the expression of tumor necrosis factor-α following stimulation with β-amyloid. These findings suggested that one common polymorphism in pri-miR-146a may contribute to the genetic predisposition to AD by disrupting the production of miR-146a-5p and affecting the expression and function of TLR2. Source


Zhang B.,Shandong University | Chen C.-F.,Shandong University | Wang A.-H.,Shanghai Fengxian Central Hospital | Lin Q.-F.,Shanghai Fengxian Central Hospital
European Review for Medical and Pharmacological Sciences | Year: 2015

OBJECTIVE: The aim of this study was to investigate the role of miR-16 in Alzheimer's disease (AD) and to explore its mechanism of action. MATERIALS AND METHODS: A cellular AD model using PC12 cells and primary hippocampal neurons was established to evaluate the expression level of miR-16. Transfection of a miR- 16 mimic and a miR-16 inhibitor were performed to explore its effect on cell apoptosis and cell viability. In addition, we carried out bioinformatics analysis, luciferase reporting gene assay, and gene expression analyses to identify the potential target of miR-16 and to verify the effect of the target gene on the cellular AD model. RESULTS: Downregulation of miR-16 was confirmed in the cellular AD model with both PC12 cells (p < 0.05) and primary hippocampal neurons (p < 0.05). Overexpression and inhibition of miR-16 in the cellular AD model with primary hippocampal neurons decreased and increased apoptosis, respectively. The gene encoding amyloid precursor protein (APP) was identified as the target gene of miR-16. Knockdown of APP in primary hippocampal neurons decreased cell apoptosis and increased cell viability in the cellular AD model. CONCLUSIONS: Our results demonstrate that downregulation of miR-16 in primary hippocampal neurons play an important role in the paracrine effect and might be involved in the development of AD. Source


He Z.,Shanghai Fengxian Central Hospital | Liu Y.,Shanghai University | Xue F.,Shanghai Fengxian Central Hospital | Xiao H.,Shanghai Fengxian Central Hospital | And 2 more authors.
Orthopedics | Year: 2012

Congenital cervical kyphosis is a rare clinical condition. The purpose of this study was to review the surgical management and outcomes of 12 consecutive cases of congenital cervical kyphosis management by the same surgical team. The authors retrospectively analyzed the records of 12 patients (5 men and 7 women) with an average age of 18.4 years (range, 15-31 years) who underwent surgery for congenital cervical kyphosis at the authors' institution between 2001 and 2005. All patients had congenital cervical kyphosis; those with secondary kyphosis deformity due to causes such as infection, tumors, and surgery were excluded. The indications for surgery were signs of spinal cord compression with progression of clinical symptoms such as decreased muscle strength and paresthesia. All patients had radiographic evidence of cervical kyphosis. Six patients underwent anterior decompression, autogenous bone grafting, and instrumentation, and the other 6 patients underwent combined anterior-posterior surgery. All surgeries were performed successfully with no complications. Bone graft fusion occurred in 11 patients. In 1 patient who underwent anterior surgery, the bone graft was partly absorbed, and pseudarthrosis was noted at 3 years postoperatively. Mean Japan Orthopaedic Association cervical myelopathy score and mean Cobb angle were significantly improved at 1 week and 1 year postoperatively compared with preoperative values. Anterior and combined anterior-posterior surgical approaches are useful for the correction of congenital cervical kyphosis. Bone graft fusion is also critical for maintaining the surgical correction. Choice of surgical methods depends on the patient's clinical condition. Source


Gao G.,Shanghai Fengxian Central Hospital | Sun Z.,Shanghai Fengxian Central Hospital | Wenyong L.,Shanghai JiaoTong University | Dongxia Y.,Shanghai JiaoTong University | And 2 more authors.
Annals of Transplantation | Year: 2015

Background: Cancer stem cell-like side population (SP) cells, which may be responsible for recurrence, tumor metastasis, and resistance to cancer therapy, have been identified and characterized in several types of cell lines from gastric cancer. However, there is no report on isolation of SP cells from human gastric cancer cell line HGC-27. This study aims to analyze the proportion of SP cells in HGC-27 cell line, differentiate SP from non-side population (NSP) cells, and determine whether the SP cells have certain biological properties of stem cells. Material/Methods: (1) HGC-27 suspension was prepared and stained with Hoechst33342 and PI for flow cytometric isolation of SP (2). Differences in proliferation and stemness-related gene expression profiles (CD133, CD44, OCT-4, MDR1, EpCAM, and ABCG2) between SP and NSP cells were detected by gastric formation assay and quantitative real- time PCR (3). Oncogenicity of SP and NSP cells was determined in nude mice in vivo. Results: (1) SP cells accounted for 0.1–1.0% of HGC-27 cells, and decreased to 0% after verapamil inhibition. Using flow cytometry, we sorted 7.5×105 SP cells and most HGC-27 cells were NSP cells (2). Gastric formation assay and MTT demonstrated that there was a significant difference in proliferation between SP and NSP cells. Gene expression analysis showed that the expression of genes was significantly higher in SP cells (3). The oncogenicity experiment in nude mice revealed that 105 SP cells were able to form tumors, which demonstrated higher tumorigenicity than non-SP cells. Conclusions: These results collectively suggested that SP cells from HGC-27 cell line have some cancer stem cell properties and could be used for studying the pathogenesis of gastric cancer, which may contribute to discovery of novel therapeutic targets. © Ann Transplant, 2015. Source

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