Shenyang, China
Shenyang, China

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Qiu J.,Liaoning Medical University | Qiu J.,463 Hospital of Chinese PLA | Li W.,General Hospital of Shenyang Military Command | Feng S.,463 Hospital of Chinese PLA | And 2 more authors.
International Journal of Molecular Medicine | Year: 2013

The aim of the present study was to investigate the neuroprotective effects of bone marrow-derived endothelial progenitor cell (EPC) transplantation against cerebral ischemia/reperfusion (I/R) injury in rats and to delineate the possible underlying mechanisms. Cerebral I/R injury was established by 2 h of middle cerebral artery occlusion (MCAO) followed by reperfusion for 24 h. EPCs were isolated from bone marrow of the donor rats, grown in conditioned medium, and characterized by flow cytometry analysis of several surface markers. Labeled EPCs (106 cells) were infused into rats at the onset of reperfusion and 12 h after reperfusion via the tail vein. Infarct volume was assessed at 24 h after reperfusion by using triphenyltetrazolium chloride (TTC) staining. The expression of cell apoptosis-related proteins including Bcl-2 and Bax was determined by western blot analysis, and the activity of caspase-3 was also measured. We evaluated the activities of some antioxidative enzymes, such as superoxide dismutase (SOD), glutathione peroxidase (GSH-PX), the non-enzymatic scavenger glutathione (GSH) and detected the content of malondialdehyde (MDA) in the ischemic penumbra. Moreover, the expression of nuclear factor-κB (NF-κB) in the ischemic regions of rats was examined by immunohistochemical staining and western blot analysis. The results showed that transplantation of EPCs significantly reduced the cerebral infarct volume, decreased caspase-3 activity, upregulated Bcl-2 expression, and downregulated the expression of Bax and NF-κB. Furthermore, reduced levels of MDA, significantly elevated activities of SOD and GSH as well as GSH-PX were also found in I/R rats transplanted with EPCs. Collectively, our data demonstrated that transplantation of bone marrow-derived EPCs exerts potent neuroprotective functions against cerebral I/R injury in rats, and the protective effects may be associated with its antioxidative and anti-apoptotic properties.


Qi X.-S.,PLA Fourth Military Medical University | Qi X.-S.,General Hospital of Shenyang Military Command | Qi X.-S.,463 Hospital of Chinese PLA | Bai M.,PLA Fourth Military Medical University | Fan D.-M.,PLA Fourth Military Medical University
World Journal of Gastroenterology | Year: 2014

Currently, nonselective β-blockers (NSBBs) are commonly used for the prevention of variceal bleeding in liver cirrhosis. The beneficial effects of NSBBs are primarily attributed to the reduction in cardiac output by blockade of β1 receptors and vasoconstriction of the splanchnic circulation by the blockade of β2 receptors. The prognostic value of occlusive portal vein thrombosis (PVT) in cirrhotic patients has been increasingly recognized. The most important risk factor for the development of PVT in liver cirrhosis is the decreased portal vein inflow velocity. Collectively, we propose that the use of NSBBs potentially increases the development of portal vein thrombosis by reducing portal vein inflow velocity. The hypothesis should be confirmed by prospective cohort studies, in which cirrhotic patients without prior PVT treated with and without NSBBs are enrolled, and the development of PVT during follow-up is compared between the two groups. Additionally, subgroup analyses should be performed according to the dosage of NSBBs and the reduction of portal inflow velocity after use of NSBBs. © 2014 Baishideng Publishing Group Inc. All rights reserved.


Wang J.,China Medical University at Heping | Chen H.,China Medical University at Heping | Fu S.,China Medical University at Heping | Xu Z.-M.,463 Hospital of Chinese PLA | And 2 more authors.
Oral Oncology | Year: 2011

Chromodomain helicase DNA-binding protein 5 (CHD5) has been found to be a candidate tumor suppressor gene (TSG) in malignant neural tumors. In mice heterozygous for chd5 deficiency, the first tumor observed was pathological squamous cell carcinoma. More than 95% of primary laryngeal cancer is squamous cell carcinoma. Thus, we explored the expression of CHD5 in 65 patients with laryngeal squamous cell carcinoma (LSCC) using real-time PCR, immunohistochemistry and Western blotting. DNA methylation was detected using bisulfate-specific sequencing. The potential function of CHD5 was determined using MTT, apoptosis and transwell migration assays in CHD5-transfected Hep-2 cells. Our results revealed that the mRNA and protein expression levels of CHD5 in LSCC tissues were significantly lower than those in clear surgical margin tissues (p < 0.05), and there is a significant correlation between the mRNA and protein expression levels of CHD5 (p < 0.01). In addition, there were significant differences in CHD5 mRNA and protein levels with respect to the patient's clinical stage (p < 0.05). Aberrant methylation of the CHD5 promoter was frequently found in the Hep-2 cell line and LSCC tumor tissues, especially tumor tissues from advanced TNM (p < 0.05) or older patients (p < 0.05). Finally, ectopic expression of CHD5 in laryngeal cancer cells led to significant inhibition of growth and invasiveness. Our data suggest that CHD5 is a tumor suppressor gene that is epigenetically downregulated in LSCC. © 2011 Elsevier Ltd.


Che X.-H.,China Medical University at Heping | Chen H.,China Medical University at Heping | Xu Z.-M.,463 Hospital of Chinese PLA | Shang C.,China Medical University at Heping | And 2 more authors.
BMC Cancer | Year: 2010

Background: 14-3-3epsilon regulates a wide range of biological processes, including cell cycle control, proliferation, and apoptosis, and plays a significant role in neurogenesis and the formation of malignant tumours. However, the exact function and regulatory mechanism of 14-3-3epsilon in carcinogenesis have not been elucidated.Methods: The expression of 14-3-3epsilon was assessed by RT-PCR and western blotting. The invasiveness and viability of Hep-2 cells were determined by the transwell migration assay and MTT assay, respectively. Cell cycle and apoptosis of Hep-2 cells were detected by flow cytometry.Results: The mRNA and protein expression of 14-3-3epsilon in larynx squamous cell carcinoma (LSCC) tissues were significantly lower than those in clear surgical margin tissues. Statistical analysis showed that the 14-3-3epsilon protein level in metastatic lymph nodes was lower than that in paired tumour tissues. In addition, the protein level of 14-3-3epsilon in stage III or IV tumours was significantly lower than that in stage I or II tumours. Compared with control Hep-2 cells, the percentages of viable cells in the 14-3-3epsilon-GFP and negative control GFP groups were 36.68 ± 14.09% and 71.68 ± 12.10%, respectively. The proportions of S phase were 22.47 ± 3.36%, 28.17 ± 3.97% and 46.15 ± 6.82%, and the apoptotic sub-G1 populations were 1.23 ± 1.02%, 2.92 ± 1.59% and 13.72 ± 3.89% in the control, negative control GFP and 14-3-3epsilon-GFP groups, respectively. The percentages of the apoptotic cells were 0.84 ± 0.25%, 1.08 ± 0.24% and 2.93 ± 0.13% in the control, negative control GFP and 14-3-3epsilon-GFP groups, respectively. The numbers of cells that penetrated the filter membrane in the control, negative control GFP and 14-3-3epsilon-GFP groups were 20.65 ± 1.94, 17.63 ± 1.04 and 9.1 ± 0.24, respectively, indicating significant differences among the different groups.Conclusions: Decreased expression of 14-3-3epsilon in LSCC tissues contributes to the initiation and progression of LSCC. 14-3-3epsilon can promote apoptosis and inhibit the invasiveness of LSCC. © 2010 Che et al; licensee BioMed Central Ltd.


Jia J.,Liaoning Medical University | Jia J.,463 Hospital of Chinese PLA | Dai S.,463 Hospital of Chinese PLA | Sun X.,463 Hospital of Chinese PLA | And 6 more authors.
Molecular Medicine Reports | Year: 2015

Human esophageal cancer-related gene 4 (ECRG4) is a potential tumor suppressor gene isolated from human esophageal epithelial cells. Studies have shown that ECRG4 effectively inhibits the proliferation of tumor cells and induces apoptosis. However, the role of ECRG4 in laryngeal cancer has not yet been clearly defined. In this study, a human laryngeal cancer cell line stably overexpressing ECRG4 was established. The effect of ECRG4 on the proliferation and apoptosis of laryngeal cancer cells and the associated mechanisms were investigated. The Hep-2 human laryngeal carcinoma cell line exhibited a low basal level of ECRG4 expression and was selected for the present study. The eukaryotic expression plasmid pcDNA3.1-ECRG4 was constructed and introduced into Hep-2 cells by transfection reagents. Western blot analysis, reverse transcription-quantitative polymerase chain reaction and immunofluorescence staining confirmed high-level expression of ECRG4. The 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and colony formation assay showed that ECRG4 overexpression suppressed the proliferative capacity of laryngeal cancer cells in vitro. Cell cycle analysis showed that ECRG4 induced cell cycle arrest at the G0/G1 phase. Flow cytometric analysis and Hoechst staining demonstrated that overexpression of ECRG4 significantly induced apoptosis. Western blot analysis confirmed that Bcl-2-associated X protein, cleaved-caspase-3 and cleaved-poly (ADP-ribose) polymerase were upregulated in the apoptotic process, whereas B-cell lymphoma 2 was downregulated. In conclusion, overexpression of ECRG4 inhibited laryngeal cancer cell proliferation and induced cancer cell apoptosis. Therefore, ECRG4 exhibits potential as an effective target in gene therapy for laryngeal cancer.


Fan M.-S.,463 Hospital of Chinese PLA | Yang X.-F.,463 Hospital of Chinese PLA | Wu Y.-X.,463 Hospital of Chinese PLA | Zhang Y.-B.,463 Hospital of Chinese PLA | And 4 more authors.
Journal of Clinical Rehabilitative Tissue Engineering Research | Year: 2011

BACKGROUND: The transplantation of bone marrow and autologous peripheral blood stem cells into the muscles of lower limbs of diabetes patients with lower limb ischemia can promote blood vessel regeneration, improve and recover the blood flow of affected limbs of diabetes patients with lower limb ischemia. However, the collection has great risk, and requires high levels in patients' age, body condition and psychological reception degree. Compared with bone marrow and autologous peripheral blood stem cells, umbilical cord resource is abundant; cell collection is simple; immunogenicity is weak. OBJECTIVE: To investigate the feasibility that transplantation of human umbilical cord mesenchymal stem cells (HUCMSCs) for treatment of diabetic rabbit of lower limbs extremity arterial. METHODS: Models of diabetic rabbits were established. Femoral arteries of lower limbs were ligated. Prepared HUCMSCs in sign of DIL were directly injected into the left hindlimb four-headed thigh muscles in diabetic rabbit models (cell quantity was 1.67×106 per point). Saline was directly injected into right lower limbs. At 2 and 4 weeks, adductor and gastrocnemius nuscle of the bilateral hindlimbs were collected. Angiogenesis of pathological sections was observed. Capillary density and skin temperature were measured using immunohistochemical staining for CD31. RESULTS AND CONCLUSION: Pathological sections exhibited that collateral vessels were significantly increased in the transplantation group compared with control group. Immunohistochemical staining for CD31 demonstrated that the capillary density was significantly higher in the transplantation group compared with control group. The skin temperature was significantly greater in the transplantation group compared with control group (P ≤ 0.05). Results suggested that HUCMSCs transplantation in the treatment of diabetic rabbit of lower limbs ischemia is an effective measure.

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