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Yin N.,Shanghai Fengxian District Central Hospital | Guan J.,Shanghai Fengxian District Central Hospital | He N.,Shanghai JiaoTong University | Xue F.,Shanghai Fengxian District Central Hospital
Chromatographia | Year: 2016

The suitability of HPLC with fluorescent derivatization for simultaneous determination of eight collagen-related bone turnover biomarkers, 4-hydroxy-l-proline (OHP), pyridinoline (PYD), hydroxylysine (Hyl), deoxypyridinoline, and their glycosides, galactosyl-Pyd (Gal-Pyd, GP), galactosyl-glucosyl-Pyd, β-1-galactosyl-O-hydroxylysine (Gal-Hyl, GH), and α-1,2-glucosyl-galactosyl-O-hydroxylysine (GGH), in diverse clinical samples was investigated. 5-Carboxyfluorescein succinimidyl ester (CFSE) was chosen as the fluorescent labeling reagent. Various parameters affecting derivatization and separation were optimized. Under optimal conditions, maximum derivatization could be achieved in 25 min at 30 °C, and complete baseline separation could be completed within 25 min, with relative standard deviation values for corrected peak areas less than 3.5 % for intraday assay and 4.2 % for interday assay, respectively. The limits of detection were determined to be 0.1–1.7 nM, which are well below the expected concentrations in the real samples. The method has been used for the quantitative determination of eight target analytes in various clinical samples, including cell cultures of normal human osteoclast, preosteoblast, osteoclastoma cells and serum and urine fluids from patients with fracture, osteitis fibrosa, or osteoclastoma as well as healthy subjects, with spiked recoveries of 96–103 %. This original application of HPLC–fluorescence detection represents a powerful tool for investigating the dynamics of metabolic imbalance of these biomarkers for bone turnover under physiological and pathophysiological conditions. © 2016, Springer-Verlag Berlin Heidelberg.

Quan Y.,Jilin University | Quan Z.,Shanghai Fengxian District Central Hospital | Zheng C.,Jilin University | Deng W.,Jilin University | And 2 more authors.
Translational Cancer Research | Year: 2015

Background: Forkhead box protein C2 (FOXC2) is an epithelial-to-mesenchymal transition (EMT)-inducing transcription factor and involved in several cancer developments. In this study, we aimed to investigate the role of FOXC2 in ovarian cancer. Methods: A total of 13 ovarian tissues (11 cases of epithelial ovarian cancer, one case of borderline serous cystadenoma and one case of normal ovarian tissue) were collected and tissue specimens were stained to detect the expression and location of FOXC2 in ovarian cancer by immunohistochemistry. Then shRNAs targeting FOXC2 mRNA were designed to construct FOXC2-shRNA-expressing lentiviral vector (LV- FOXC2). Real-time RT-PCR and western blot assays were conducted to examine the interference effects of shRNA to FOXC2. Additionally, cell counting kit-8 (CCK8) was performed to detect cell proliferation of ID8 cells (mouse ovarian cancer cell) infected with FOXC2-shRNA-lentivirus. Results: No expression of FOXC2 was found in normal ovarian tissue and a large number of FOXC2 positive cells were found in borderline serous cystadenoma and ovarian cancer tissues including vascular endothelial cells, and FOXC2 was mainly seen in nucleus of ovarian cancer cells. Interference lentivirus of FOXC2 was successfully constructed with the virus titers of 1.2×108~1.3×108 Tu/mL, which significantly decreased the mRNA and protein expression levels of FOXC2 (P < 0.05). Furthermore, cell proliferation was significantly suppressed with a time-dependent effect in ID8 cells infected with FOXC2-shRNA-lentivirus. Conclusions: This study might help to understand the molecular mechanism of FOXC2 in ovarian cancer and provide theoretical foundation for the development of diagnosis and treatment for ovarian cancer by targeting FOXC2. © 2011 - 2016 Translational Cancer Research. All rights reserved.

Liu F.,Shanghai Fengxian District Central Hospital | Wang J.,Shanghai JiaoTong University | Fu Q.,Shanghai JiaoTong University | Zhang X.,Shanghai JiaoTong University | And 4 more authors.
Tumor Biology | Year: 2015

Cisplatin is a commonly used chemotherapy drug for prostate cancer (PC). However, some PCs are resistant to cisplatin treatment, while the molecular mechanisms underlying the resistance of PCs to cisplatin are not completely understood. In this study, we found that cisplatin dose-dependently activated Beclin-1 in two PC cell lines, PC3 and LNCap. Autophagy suppression significantly increased the cisplatin-induced cell death of these PC cells in a CCK-8 assay. Moreover, microRNA (miR)-144 levels were significantly downregulated in cisplatin-treated PC cells, in a VEGF-dependent manner. Bioinformatics analysis showed that miR-144 targeted the 3′-UTR of Beclin-1 mRNA to inhibit its translation, which was confirmed by luciferase reporter assay. In PC patients after cisplatin treatment, low miR-144 levels appeared to predict poor outcome of patients’ survival. Together, these data suggest that cisplatin may induce VEGF to suppress miR-144 levels in PC cells, which subsequently upregulates Beclin-1 to increase autophagic cell survival against cisplatin-induced cell death. Upregulation of miR-144 or suppression of cell autophagy may improve the outcome of cisplatin therapy in PC. © 2015 International Society of Oncology and BioMarkers (ISOBM)

Wen S.-Y.,Shanghai JiaoTong University | Lin Y.,Shanghai JiaoTong University | Yu Y.-Q.,Shenyang Medical College | Cao S.-J.,Shanghai Songjiang District Central Hospital | And 11 more authors.
Oncogene | Year: 2014

Although significant advances have recently been made in the diagnosis and treatment of cervical carcinoma, the long-term survival rate for advanced cervical cancer remains low. Therefore, an urgent need exists to both uncover the molecular mechanisms and identify potential therapeutic targets for the treatment of cervical cancer. MicroRNAs (miRNAs) have important roles in cancer progression and could be used as either potential therapeutic agents or targets. miR-506 is a component of an X chromosome-linked miRNA cluster. The biological functions of miR-506 have not been well established. In this study, we found that miR-506 expression was downregulated in approximately 80% of the cervical cancer samples examined and inversely correlated with the expression of Ki-67, a marker of cell proliferation. Gain-of-function and loss-of-function studies in human cervical cancer, Caski and SiHa cells, demonstrated that miR-506 acts as a tumor suppressor by inhibiting cervical cancer growth in vitro and in vivo. Further studies showed that miR-506 induced cell cycle arrest at the G1/S transition, and enhanced apoptosis and chemosensitivity of cervical cancer cell. We subsequently identified Gli3, a hedgehog pathway transcription factor, as a direct target of miR-506 in cervical cancer. Furthermore, Gli3 silencing recapitulated the effects of miR-506, and reintroduction of Gli3 abrogated miR-506-induced cell growth arrest and apoptosis. Taken together, we conclude that miR-506 exerts its anti-proliferative function by directly targeting Gli3. This newly identified miR-506/Gli3 axis provides further insight into the pathogenesis of cervical cancer and indicates a potential novel therapeutic agent for the treatment of cervical cancer.Oncogene advance online publication, 10 March 2014; doi:10.1038/onc.2014.9.

He Z.,Shanghai Fengxian District Central Hospital | Zhu H.,Shanghai Fengxian District Central Hospital | Ding L.,Shanghai Fengxian District Central Hospital | Xiao H.,Shanghai Fengxian District Central Hospital | And 2 more authors.
Genetics and Molecular Research | Year: 2013

Ossification of the posterior longitudinal ligament (OPLL) is a condition of the spine that can cause paralysis by compressing the spinal cord. The aim of this study was to evaluate the possible role of nucleotide pyrophosphatase phosphodiesterase 1 gene (NPP1) polymorphism in the etiology and pathology of the OPLL in Chinese patients. DNA from patients with OPLL (N = 95) and without OPLL (N = 90) were genotyped for 4 NPP1 single-nucleotide polymorphisms (SNPs): A533C, C973T, IVS15-14T→C, and IVS20-11delT. An association study evaluated the relationship between specific SNP genotypes and susceptibility. We also evaluated whether genotypes of these SNPs were associated with disease severity and the probability of disease progression after surgery. The C973T and IVS15-14T SNPs were associated with the existence of the disease. The TT genotypes of C973T and IVS15→14T as well wild-type IVS20 (lack of deletion) were associated with more severe disease. Patients with the T deletion of IVS20 or the AA genotype of A533C had an approximately 3 times greater chance of not having than having disease progression after surgery. We concluded that the 4 SNPs analyzed appeared to have different effects on the etiology and pathology of OPLL. To our knowledge, this study is the first to investigate the relationship between these SNPs and disease progression after surgery. Our findings suggest that the presence of specific genotypes of the IVS20-11delT and A533C SNPs may predict disease outcome after surgical intervention. © FUNPEC-RP.

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