3.Construction and identification of recombinant lentivirus vector for microRNA-223 overexpression and suppression.
Yun WANG ; Ning JI ; Min ZHOU ; Lu JIANG ; Qianming CHEN
West China Journal of Stomatology 2015;33(5):451-455
OBJECTIVETo construct microRNA-223 overexpression and suppression lentivirus vectors and determine their effects after infecting oral squamous cell carcinoma (OSCC) cell line.
METHODSLentivirus vectors GV229 and GV232 were cut by the restriction sites of Age I and EcoR I and connected to the target gene, which contained mature microRNA-223 and microRNA-223 oligonucleotide. Real-time polymerase chain reaction (PCR) method was used to detect the microRNA-223 expression level after infecting the recombinant lentivirus vector into the OSCC cell line.
RESULTSThe successful construction of microRNA-223 recombinant lentivirus vectors was confirmed by the PCR method and DNA sequencing. HN-30 cell infected with microRNA-223 overexpression vector showed a significant increased in microRNA-223 expression, whereas HN-30 cell infected with microRNA-223 inhibitor vector suppressed microRNA-223 expression.
CONCLUSIONThe microRNA-223 overexpression and suppression lentivirus vectors are successfully constructed. These vectors could alter the expression level of microRNA-223 in OSCC cell line significantly, and provide a stable cell line for functional studies in the future.
Base Sequence ; Cell Line, Tumor ; Genetic Vectors ; Humans ; Lentivirus ; MicroRNAs ; metabolism ; Real-Time Polymerase Chain Reaction
4.Influence of lead on expression of epithelial mesenchymal transitions and fibrosis related factors of HK-2 cells.
Gui-Feng ZHOU ; Yun-Sheng JIANG ; You-Ming PENG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2008;26(10):621-623
Cell Differentiation
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drug effects
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Cell Line
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Connective Tissue Growth Factor
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metabolism
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Epithelial Cells
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cytology
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drug effects
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metabolism
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Epithelial-Mesenchymal Transition
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drug effects
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Humans
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Kidney Tubules, Proximal
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cytology
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Lead
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toxicity
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Transforming Growth Factor beta
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metabolism
5.Preventing inflammatory reaction with estrogen after cerebral ischemia/reperfusion injury
Yun JIANG ; Guanghui CHEN ; Guoqing ZHOU ; Kui MENG
Chinese Journal of Tissue Engineering Research 2006;10(46):214-215
BACKGROUND:Leukocytic infiltration induced by release of inflammatory cytotkines and up-regulation of adhesion molecules is closely associated with the formation of cerebral infarcted focus. The related factors have been widely studied.OBJECTIVE: To explore influence of estrogen on inflammatory reaction in rats after focal ischemia/reperfusion injury.DESIGN: Randomized controlled experiment.SETTING: Nanjing General Hospital, Nanjing Military Area Command of Chinese PLA.MATERIALS:The experiment was performed at the Department of Neurology, Department of Pathology, Nanjing General Hospital, Nanjing Military Area Command of Chinese PLA. Adult male SD rats were selected to establish cerebral ischemic models, with the body mass of 280-350 g.METHODS:The rats were assigned into 3 groups: control group,ovariectomized group and estrogen treatment group (estradiol, 200 μg/kg,subcutaneous injection, once a week for 4 weeks). Four weeks later, models with right middle cerebral artery occlusion (MCAO) for 1 hour and 2 hours as well as reperfusion for 0, 1, 3, 6, 22 and 70 hours were established with thread embolism method. Mean number of infiltrative neutrophils in brain tissue was calculated under microscope with 10 high power fields in ischemic hemisphere with hematoxylin and eosin staining. Expression of nuclear factor-κB was determined with immunohistochemical method.MAIN OUTCOME MEASURES:Infiltration of neutrophils and expression of nuclear factor-κB in brain parenchyma.RESULTS: ①Expression of nuclear factor-κB: There was expression of nuclear factor-κB in the ovariectomized group at hour 1 after ischemia.Positive cells appeared at hour 2 after ischemia in the control group and estrogen treatment group. The expression was in the peak at ischemia for 2hours and reperfusion for 3 hours in the three groups, and decreased gradually. There was slight expression at reperfusion for 70 hours in the ovariectomized group, while there was no clear factor-κB positive cell at reperfusion for 22 hours in the control group and estrogen treatment group.②Infiltrative neutrophils in cerebral ischemic region in the ovariectomized group significantly increased at ischemia for 2 hours and reperfusion for 22 hours. Compared with the estrogen treatment group, there was significant difference (P=0.045). At ischemia for 2 hours and reperfusion for 70 hours,infiltrative neutrophils in the ovariectomized group were more than those in the control group and estrogen treatment group, but there were significant differences only between ovariectomized group and control group.CONCLUSION: Estrogen can inhibit inflammatory reaction after cerebral ischemia/reperfusion injury.
6.Feasibility of two kinds of atlantal lateral mass screw fixation
Min GONG ; Junwei JIANG ; Hao LIU ; Yun ZHOU
Chinese Journal of Tissue Engineering Research 2014;(31):4968-4972
BACKGROUND:Atlantal lateral mass screw fixation contains lateral mass screw fixation under the posterior arch of the atlas and lateral mass screw fixation via the posterior arch of atlas (also cal ed atlas pedicle screw fixation) according to different insertion points. The two methods have their advantages and disadvantages. There are lacks of the comparative studies on bony anatomy feasibility by two kinds of atlantal lateral mass screw fixation.
OBJECTIVE:To compare the feasibility associated with two kinds of atlantal lateral mass screw fixations taking Chinese data of atlantal bony anatomy as evidence.
METHODS:Data of CT scans in 30 adults with cervical spondylosis (60 sides) were col ected, and data were reconstructed using CT workstation. We measured key bone anatomy structure after atlantal lateral mass screw fixation. The atlantal lateral mass was suitable for lateral mass screw fixation via the posterior arch of atlas if its height and width of vertebral artery groove at the posterior arch ≥ 4 mm. It was suitable for lateral mass screw fixation under the posterior arch of the atlas if the height of posterior arch of atlas lateral mass ≥ 4 mm.
RESULTS AND CONCLUSION:The height of vertebral artery groove at the posterior arch was (4.54±1.17) mm. The width of vertebral artery groove at the posterior arch was (8.69±1.12) mm. The height of posterior arch of atlas lateral mass was (4.98±1.07) mm. There were 41 cases with the height of vertebral artery groove at the posterior arch>4 mm (suitable for lateral mass screw fixation via the posterior arch of atlas, occupying 68%. There were 52 cases with the height of posterior arch of atlas lateral mass>4 mm (suitable for lateral mass screw fixation under the posterior arch of the atlas), occupying 87%. There were significant differences in the differences between the two groups (P<0.05). These results indicated that lateral mass screw fixation under the posterior arch of the atlas was more feasible compared with lateral mass screw fixation via the posterior arch of atlas. Preoperative CT measure for key anatomic structure was significant for making personalized surgical plan.
7.Value of DWI for early assessment of response to concurrent chemoradiotherapy for esophageal cancer:a preliminary study
Wei CHEN ; Shengli ZHOU ; Chongchang MIAO ; Xiaodong JIANG ; Yun QIAO
Chinese Journal of Radiation Oncology 2014;23(4):312-316
Objective To investigate the feasibility and value of diffusion-weighted imaging (DWI) in the prediction and early assessment of response to concurrent chemoradiotherapy (CCRT) for esophageal cancer.Methods A total of 40 patients with pathologically confirmed esophageal cancer who received CCRT were included in the study.Routine 3.0 T MRI and DWI were performed at different time points of treatment.The RECIST standard was adopted to evaluate short-term outcomes and divide the patients into remission group (complete remission and partial remission) and non-remission group (stable disease and progressive disease).Group t-test was used for between-group comparison.The receiver operating characteristic (ROC) curve was used to analyze the change rates of apparent diffusion coefficient (ADC) value at different time points of treatment.Results There were 30 patients in the remission group and 10 patients in the non-remission group.The remission group had a significantly higher increase in ADC value than the non-remission group by the end of the first week of treatment (P =0.000).The maximum diameters of tumors for the emission group and non-remission group at the end of the first week of treatment were not significantly different from those before treatment (66.10 mm vs.62.63 mm,P =0.407 ; 70.90 mm vs.68.30 m,P =0.552).The ADC value before treatment had a negative correlation with the reduction rate of the maximum diameter of tumor (r =-0.680,P =0.000).The area under the ROC curve was the largest at the end of the first week of treatment (Az =0.783).If using 15.5 % increase in ADC value by the end of the first week as the threshold value for evaluating tumor response,the sensitivity,specificity,positive predictive value,and negative predictive value were 86.7%,70.0%,89.7%,and 63.6%,respectively.Conclusions DWI can be used as a new imaging method for the prediction and early assessment of the response to CCRT for esophageal cancer.The change rate of ADC value by the end of the first week of treatment is sensitive in assessing treatment response,so ADC value can be monitored at this time point.
9.The effect of combined application of low dose fentanyl and midazolam on sodium channels in rat cerebral cortical neurons.
Yun-Chun YANG ; Xian ZHOU ; Jia-Li WU ; Xuan JIANG ; Shu-Zhi ZHOU ; Xiao-Bin WANG
Chinese Journal of Applied Physiology 2011;27(1):85-87
Anesthetics, Intravenous
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administration & dosage
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pharmacology
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Animals
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Animals, Newborn
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Cerebral Cortex
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cytology
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metabolism
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Drug Synergism
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Female
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Fentanyl
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administration & dosage
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pharmacology
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Male
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Midazolam
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administration & dosage
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pharmacology
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Neurons
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metabolism
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Patch-Clamp Techniques
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Primary Cell Culture
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Rats
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Rats, Sprague-Dawley
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Voltage-Gated Sodium Channels
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drug effects
10.Inhibitory effect of kaempferol on inflammatory response of lipopolysaccharide-stimulated human mast cells.
Yun-jiang ZHOU ; Hu WANG ; Li LI ; He-huan SUI ; Jia-jun HUANG
Acta Pharmaceutica Sinica 2015;50(6):702-707
This study is to investigate the inhibitory effect of kaempferol on inflammatory response of lipopolysaccharide(LPS)-stimulated HMC-1 mast cells. The cytotoxicity of kaempferol to HMC-1 mast cells were analyzed by using MTT assay and then the administration concentrations of kaempferol were established. Histamine, IL-6, IL-8, IL-1β and TNF-α were measured using ELISA assay in activated HMC-1 mast cells after incubation with various concentrations of kaempferol (10, 20 and 40 µmol.L-1). Western blot was used to test the protein expression of p-IKKβ, IκBα, p-IκBα and nucleus NF-κB of LPS-induced HMC-1 mast cells after incubation with different concentrations of kaempferol. The optimal concentrations of kaempferol were defined as the range from 5 µmol.L-1 to 40 µmol.L-1. Kaempferol significantly decreased the release of histamine, IL-6, IL-8, IL-1β and TNF-α of activated HMC-1 mast cells (P<0.01). After incubation with kaempferol, the protein expression of p-IKKβ, p-IKBa and nucleus NF-κB (p65) markedly reduced in LPS-stimulated HMC-1 mast cells (P<0.01). Taken together, we concluded that kaempferol markedly inhibit mast cell-mediated inflammatory response. At the same time, kaempferol can inhibit the activation of IKKβ, block the phosphorylation of IκBα, prevent NF-KB entering into the nucleus, and then decrease the release of inflammatory mediators.
Cells, Cultured
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Histamine
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metabolism
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Humans
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I-kappa B Kinase
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metabolism
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I-kappa B Proteins
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metabolism
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Inflammation
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metabolism
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Interleukin-1beta
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metabolism
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Interleukin-6
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metabolism
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Interleukin-8
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metabolism
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Kaempferols
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pharmacology
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Lipopolysaccharides
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Mast Cells
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drug effects
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NF-KappaB Inhibitor alpha
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NF-kappa B
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metabolism
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Tumor Necrosis Factor-alpha
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metabolism