Fifth Central Hospital of Tianjin
Fifth Central Hospital of Tianjin
Jiang M.,Duke University |
Yu S.,Duke University |
Yu S.,Nantong University |
Yu Z.,Duke University |
And 8 more authors.
Stroke | Year: 2017
Background and Purpose-Impaired protein homeostasis induced by endoplasmic reticulum dysfunction is a key feature of a variety of age-related brain diseases including stroke. To restore endoplasmic reticulum function impaired by stress, the unfolded protein response is activated. A key unfolded protein response prosurvival pathway is controlled by the endoplasmic reticulum stress sensor (inositol-requiring enzyme-1), XBP1 (downstream X-box-binding protein-1), and O-GlcNAc (O-linked β-N-Acetylglucosamine) modification of proteins (O-GlcNAcylation). Stroke impairs endoplasmic reticulum function, which activates unfolded protein response. The rationale of this study was to explore the potentials of the IRE1/XBP1/O-GlcNAc axis as a target for neuroprotection in ischemic stroke. Methods-Mice with Xbp1 loss and gain of function in neurons were generated. Stroke was induced by transient or permanent occlusion of the middle cerebral artery in young and aged mice. Thiamet-G was used to increase O-GlcNAcylation. Results-Deletion of Xbp1 worsened outcome after transient and permanent middle cerebral artery occlusion. After stroke, O-GlcNAcylation was activated in neurons of the stroke penumbra in young mice, which was largely Xbp1 dependent. This activation of O-GlcNAcylation was impaired in aged mice. Pharmacological increase of O-GlcNAcylation before or after stroke improved outcome in both young and aged mice. Conclusions-Our study indicates a critical role for the IRE1/XBP1 unfolded protein response branch in stroke outcome. O-GlcNAcylation is a prosurvival pathway that is activated in the stroke penumbra in young mice but impaired in aged mice. Boosting prosurvival pathways to counterbalance the age-related decline in the brain's self-healing capacity could be a promising strategy to improve ischemic stroke outcome in aged brains. © 2017 American Heart Association, Inc.
Lan F.,Tianjin Hospital |
Yang Y.,Tianjin Hospital |
Han J.,Fifth Central Hospital of Tianjin |
Wu Q.,Tianjin Huanhu Hospital |
And 2 more authors.
International Journal of Oncology | Year: 2016
The survival benefits of patients with glioblastoma (GBM) remain unsatisfactory due to the intrinsic or acquired resistance to temozolomide (TMZ). We elucidated the mechanisms of sulforaphane (SFN) reverse TMZ resistance in TMZ-inducing cell lines by inhibiting nuclear factor-κB (NF-κB) transcriptional activity. TMZ-resistant cell lines (U87-R and U373-R) were generated by stepwise (6 months) exposure of parental cells to TMZ. Luciferase reporter assay, biochemical assays and subcutaneous tumor establishment were used to characterize the antitumor effect of SFN. MGMT expression and 50% inhibiting concentration (IC50) values of TMZ in GBM cell lines were assessed. Next, we established that U87-R and U373-R cells presenting high IC50 of TMZ, activated NF-κB transcription and significantly increased MGMT expression compared with untreated cells. Furthermore, we revealed that SFN could significantly suppress proliferation of TMZ-resistant GBM cells. In addition, SFN effectively inhibited activity of NF-κB signaling pathway and then reduced MGMT expression to reverse the chemo-resistance to TMZ in T98G, U87-R and U373-R cell lines. Sequential combination with TMZ synergistically inhibited survival capability and increased the induction of apoptosis in TMZ-resistant GBM cells. Finally, a nude mouse model was established with U373-R cell subcutaneous tumor-bearing mice, and results showed that SFN could remarkably suppress cell growth and enhance cell death in chemo-resistant xenografts in the nude mouse model. Collectively, the present study suggests that the clinical efficacy of TMZ-based chemotherapy in TMZ-resistant GBM may be improved by combination with SFN.
Zhu B.,Fifth Central Hospital of Tianjin |
Li Y.,Zhejiang University |
Li M.,Zhejiang University |
Yang X.,Jilin University |
And 3 more authors.
Spinal Cord | Year: 2013
Study design:Spinal cord injury (SCI) is a devastating and common neurologic disorder that has profound influences on modern society from physical, psychosocial and socio-economic perspectives.Objectives:To analyze the dynamic changes in protein expression during SCI after ischemia-reperfusion. Methods:We used two-dimensional difference gel electrophoresis combined with matrix-assisted laser desorption/ionization time-of-flight/time-of-flight MS to give a global analysis of protein dynamic change during SCI after ischemia-reperfusion. Dynamic changes in protein expression were investigated from 6 to 48 h in SCI after ischemia-reperfusion using a proteomics tool.Results:Twenty-one proteins were identified in total, including neuronal proteins, glycometabolism enzymes, stress-related proteins and cytoskeleton-related proteins. These were divided into upregulated and downregulated groups. Results identified 24 h as a key time point when all proteins were changed dramatically. In addition, changes in Fascin expression were discovered in SCI for the first time.Conclusion:In conclusion, we observed dynamic proteome change correlated with SCI by ischemia-reperfusion, and provided a clue to this pathological mechanism by protein identification and analysis.
Li Y.,Fifth Central Hospital of Tianjin |
Liu H.,Fifth Central Hospital of Tianjin |
Shao J.,Fifth Central Hospital of Tianjin |
Xing G.,Fifth Central Hospital of Tianjin
Molecular Medicine Reports | Year: 2017
Gastric cancer (GC) is one of the most common types of malignancy worldwide, with high morbidity and mortality rates. The dysregulation of microRNAs (miRs) has been found to be involved in the carcinogenesis of GC. The present study aimed to investigate the underlying association between GC and miR-320a. Analysis using reverse transcription quantitative polymerase chain reaction indicated that the expression of miR-320a was downregulated and the expression of RAB14 was upregulated in GC tissues and cells, compared with the corresponding controls. MTT, colony formation assays, and flow cytometric analyses were used to evaluate the effect of miR-320a on cell proliferation and the cell cycle. The ectopic expression of miR-320a using miR-320a mimics suppressed cell viability, inhibited G1/S transition, and induced apoptosis in AGS and MKN45 cells. In addition, RAB14 was identified as a direct target gene of miR-320a, according to the results of bioinformatics analysis and a luciferase reporter assay. Downregulation of RAB14 by RAB14-small interfering RNA inhibited the viability of GC cells, which was similar to the phenotype of miR-320a mimics. Furthermore, the reintroduction of RAB14 partially abrogated the miR-320a-mediated downregulation of RAB14 and rescued the miR-320a-induced effects on GC cell growth. These findings suggest a potential novel therapeutic target for the treatment of GC.
Sun L.-L.,Fifth Central Hospital of Tianjin |
Wang Z.-X.,Fifth Central Hospital of Tianjin
Chinese Journal of Tissue Engineering Research | Year: 2017
BACKGROUND: At present, bone substitute filling is mainly used for bone defect repair. In order to understand the effect on bone defect repair, it is necessary to look into the microstructure changes of bone defects after bone substitute implantation. Synchrotron radiation-based micro-computed-tomography (SR-μCT) can be used to make high-resolution, high-resolution three-dimensional imaging without slicing and dyeing, and has high scientific and clinical value. OBJECTIVE: To explore the feasibility of SR-μCT in micron-level bone osseointegration examination in oral medicine. METHODS: This randomized controlled animal experiment was completed at the Fifth Central Hospital of Tianjin, Tianjin, China. A rabbit model of mandibular defect was made in 24 male New Zealand white rabbits. The model rats were randomly divided into four groups and received autologous bone, Bio-oss bone meal, 4-tricalcium phosphate powder and no implantation (negative control group) in the defective area, respectively. Bone samples, including the defect area and the surrounding normal bone tissue, were taken at 2, 4, 8 weeks postoperatively for SR-μCT examination, followed by histopathological examination, in order to observe the repairing effects of different types of bone implant materials from different angles. The study protocol has been approved by the Ethics Committee of the Fifth Central Hespital of Tianjin in China. The study procedures were completed in accordance with the Guidance Suggestions for the Care and Use of Experimental Animals of China and the guidelines of the National Institutes of Health, USA. RESULTS AND CONCLUSION: In this study, SR-μCT could be used to observe the bone microstructure and osseointegration with no damage to samples to collect accurate quantitative data, including bone volume, number of bone trabeculae and bone mineral density. Therefore, SR-μCT can fully analyze the biocompatibility of bone implant material in vivo, give insight into the micron-level changes of different types of bone implant materials in the bone defect, thereby providing experimental evidence to improve bone defect healing. © 2017, Journal of Clinical Rehabilitative Tissue Engineering Research. All rights reserved.
Liu Z.,Fifth Central Hospital of Tianjin |
Jiang Z.,Fifth Central Hospital of Tianjin |
Huang J.,Duke University |
Huang S.,Duke University |
And 7 more authors.
International Journal of Oncology | Year: 2014
Epidermal growth factor receptor (EGFR) signaling regulates glioblastoma cell proliferation, survival, migration and invasion and plays a key role in tumor progression. We show that microRNA-7 (miR-7) is a common regulator of the phosphoinositide-3-kinase (PI3K)/ATK and Raf/mitogen-activated protein kinase kinase (MEK)/extracellular signal-regulated kinase (ERK) pathways, both of which are launched by EGFR through its two direct targets, the transcription factors PI3K and Raf-1, respectively. Enforced expression of miR-7 markedly decreased expression of PI3K, phosphorylated Akt, Raf-1, phosphorylated MEK 1/2, and cyclin D1, as well as slightly reduced expression of EGFR. Forced expression of PI3K or Raf-1 transcripts lacking the 3′-untranslated region (3′-UTR) partially reversed the effects of miR-7 on cell growth inhibition and cell cycle arrest in glioma cells. Additionally, transient expression of miR-7 in glioblastoma cells strongly inhibited in vivo glioblastoma xenograft growth. We conclude that miR-7 is a potential tumor suppressor in glioblastoma that acts by targeting multiple oncogenes related to the downstream pathway of EGFR and may serve as a novel therapeutic target for malignant gliomas.
Yang W.,Duke University |
Sheng H.,Duke University |
Thompson J.W.,Duke University |
Zhao S.,Fifth Central Hospital of Tianjin |
And 5 more authors.
Stroke | Year: 2014
BACKGROUND AND PURPOSE - : Small ubiquitin-like modifier (SUMO) conjugation is a post-translational modification associated with many human diseases. Characterization of the SUMO-modified proteome is pivotal to define the mechanistic link between SUMO conjugation and such diseases. This is particularly evident for SUMO2/3 conjugation, which is massively activated after brain ischemia/stroke, and is believed to be a protective response. The purpose of this study was to perform a comprehensive analysis of the SUMO3-modified proteome regulated by brain ischemia using a novel SUMO transgenic mouse. METHODS - : To enable SUMO proteomics analysis in vivo, we generated transgenic mice conditionally expressing tagged SUMO1-3 paralogues. Transgenic mice were subjected to 10 minutes forebrain ischemia and 1 hour of reperfusion. SUMO3-conjugated proteins were enriched by anti-FLAG affinity purification and analyzed by liquid chromatography-tandem mass spectrometry. RESULTS - : Characterization of SUMO transgenic mice demonstrated that all 3 tagged SUMO paralogues were functionally active, and expression of exogenous SUMOs did not modify the endogenous SUMOylation machinery. Proteomics analysis identified 112 putative SUMO3 substrates of which 91 candidates were more abundant in the ischemia group than the sham group. Data analysis revealed processes/pathways with putative neuroprotective functions, including glucocorticoid receptor signaling, RNA processing, and SUMOylation-dependent ubiquitin conjugation. CONCLUSIONS - : The identified proteins/pathways modulated by SUMOylation could be the key to understand the mechanisms linking SUMOylation to neuroprotection, and thus provide new promising targets for therapeutic interventions. The new transgenic mouse will be an invaluable platform for analyzing the SUMO-modified proteome in models of human disorders and thereby help to mechanistically link SUMOylation to the pathological processes. © 2014 American Heart Association, Inc.
Liu X.-Z.,Fifth Central Hospital of Tianjin
Chinese Journal of Oncology | Year: 2012
Objective: To explore if folic acid/polyamide-amine (FA/PAMAM) enhances the therapeutic effect of miR-7gene therapy for glioma in vivo. Methods: The miR-7 gene was transfected into U251 glioma cells by FA/PAMAM. The efficiency of gene transfection was assessed by fluorescence microscopy. The miR-7 level was detect by quantitative RT-PCR. Intracranial glioma models were established in thymectomized mice, and FA/PAMAM nanoparticles were transplanted into the tumors in situ 3 days later. The animal survival was recorded and the gross tumor volume and degree of edema were observed by MRI. Apoptosis in the glioma cells and expression of proliferating cell nuclear antigen(PCNA), matrix metalloproteinases 2 and 9 (MMP-2 and MMP-9) were assessed by immunohistochemistry, and EGFR and AKT-2 protein expression was detected by Western blot assay. Results: Compared with the liposomes, the FA/PAMAM nanoparticles were more efficient to transfer miR-7 gene into U251 glioma cells, MRI showed that the tumor growth was much slower in the FA/PAMAM/miR-7 group, and the animal survival time was longer. The apoptosis rate was (5.3 ± 0.9) % in the control group, (11.4 ± 2.4) % in the liposome/miR-7 group, and (17.7 ± 3.7) % in the FA/PAMAM/miR-7 group. The immunohistochemical assay showed that the levels of PCNA, MMP-2 and MMP-9 protein in the FA/PAMAM/miR-7 group were (34.6 ± 5.4) %, (24.5 ± 4.1) %, (25. 4 ± 5.1) % , respectively, significantly lower than those in the liposome/miR-7group(49.3 ± 5.9)%, (31.7 ± 7.1)% and (39.4 ±6.4)%, respectively, and those in the control group (57.3 ± 7.4)%, (45.4 ± 6.9)% and (55.1 ± 7.3) %, respectively(all P < 0.05). The expressions of EGFR and AKT-2 proteins were 1.09 ± 0.12 and 0.62 ± 0.10 in the control group, 0.63 ± 0.11 and 0.43 ± 0.07 in the liposome/miR-7 group, and significantly deceased (0.47 ± 0.09 and 0.31 ± 0.04, respectively) in the FA/PAMAM/miR-7 group (all P < 0.05). Conclusion: Compared with the liposomes, FA/PAMAM can transfect miR-7 into glioma cells with a higher efficiency in vivo, makes a longer time of the drug action, and shows a certain inhibitory effect on the growth of glioma, therefore, might become a new drug targeting agent in gene therapy forglioma.
Chen H.,Tianjin Medical University |
Chen H.,Fifth Central Hospital of Tianjin |
Wang Y.,Tianjin Medical University |
Xue F.,Tianjin Medical University
Oncology Reports | Year: 2013
To determine the role played by the Wnt/β-catenin signaling pathway in the development of endometrial cancer (EC), we examined the expression of Wnt10a and Wnt10b in EC tissues and the correlation between their expression. Furthermore, the associations between these two proteins and the clinicopathological characteristics and prognosis of EC were also evaluated. In our search of alternative mechanisms, we investigated the impact of Wnt10b on proliferation and apoptosis of EC cells. Western blotting, 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and flow cytometry were used to evaluate the expression of Wnt10b and some key proteins of the Wnt/β-catenin pathway, proliferation and apoptosis in EC. Our results showed that Wnt10b expression in EC tissues was significantly higher compared to that in hyperplastic and normal samples. The expression of Wnt10a in endometrioid cancer tissues was higher compared to that in other types of cancerous samples. The difference in Wnt10b levels was significant among subgroups for histological type, grade of differentiation, FIGO phase and lymphovascular metastasis. Furthermore, no correlation was observed between the expression of Wnt10a and Wnt10b. In the follow-up, Wnt10b gene expression was frequently upregulated in EC and associated with better prognostic clinicopathological markers in EC patients. Collectively, the in vitro data showed that the upregulated expression of Wnt10b in Ishikawa cells promoted proliferation and inhibited apoptosis through β-catenin and c-myc activation and adenomatous polyposis coli (APC) inhibition, which suggests that Wnt10b activates EC via the Wnt/β-catenin pathway. These results suggest that Wnt10b likely plays an important role in the development of EC. Furthermore, these results identify a role for Wnt10b in EC cells through promoting proliferation and inhibiting apoptosis, primarily through the activation of the Wnt/β-catenin pathway. The role played by Wnt10a in EC, however, still requires further investigation.
Li A.,Fifth Central Hospital of Tianjin |
Cao X.,Tianjin Medical University
Chinese Journal of Clinical Oncology | Year: 2013
The clinical classification of metastatic spinal tumors is based on the degree of malignancy, nervous system function, osteoclasia, and prognosis of patients. The application of this classification system is relatively simple. The assessment and evaluation of patients to estimate the period of survival can guide the individualized treatment of patients. Patients with clear surgical indications can choose to undergo a palliative operation, tumor resection, or total spondylectomy. Alternatively, other patients can opt to have minimally invasive surgery aside from the normal surgical approach. Stereotactic radiotherapy and intensity-modulated radiation therapy combined with the use of radiofrequency ablation, radiation, and other therapies can achieve local tumor control, relieve pain, and maintain normal neurological function. Thus, the quality of life of the patient is improved. Percutaneous vertebroplasty, radiofrequency ablation, spinal endoscopy, and other modern forms of minimally invasive surgery will have more applications in the treatment of spinal metastatic tumors.