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Objective:To investigate the effects of microRNA(miR)-133b on epithelial-mesenchymal transition(EM)of human small airway epithelial cells induced by cigarette smoke extracts(CSE)and its regulatory mechanisms.Methods:The miR-expression profiles with microarray in airway epithelial cells of patients with chronic obstructive pulmonary disease were searched in the Gene Expression Omnibus(GEO)database, and the differentially expressed miRs were searched and verified by a real-time fluorescence quantitative method(qRT-PCR). Human small airway epithelial cells(HSAEpiC)were divided into the control group, the CSE group, the CSE+ miR-133b inhibitor transfection group(inhibitor group)and the CSE+ miR-133b inhibitor negative control transfection group(inhibitor control group)according to different intervention methods.Levels of miR-133b and mRNA levels of transforming growth factor(TGF)-β1, Smad2, E-cadherin and vimentin were detected by RT-PCR; Protein levels of E-cadherin and vimentin were detected by enzyme-linked immunosorbent assays(ELISA)and Western blotting.Results:Nine differentially expressed miRs were found in GSE53519, with miR-133b showing the most significant differential in thee HSAEpiC cell model after verification.CSE induced morphological changes in HSAEpiC cells, and miR-133b inhibitors could partially reverse the morphological changes in cell mode.mRNA and protein expressions of E-cadherin were decreased and expression of Vimentin mRNA and protein were increared in CSE induced HSAEpiC cells( F=9.09、12.35、7.57、101.87, P=0.015、0.007、0.023、0.000); miR-133b inhibitors partally reversed the mRNA and protein expressions of E-cadherin and Vimentin( F=40.59、27.74、15.87、20.42, P=0.000、0.001、0.004、0.002). CSE induced incresed expression of TGF-β1 mRNA and Smad mRNA in HSAEpiC cells, and miR-133b inhibitors partially reversed the changes in TGF-β1 mRNA and Smad mRNA( F=17.25、64.15, P=0.003、0.000). Conclusions:miR-133b may regulate CSE-related HSAEpiC cell EMT through the TGF-β1/Smad pathway.
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Objective To investigate the effect of hyperoside on proliferation and killing activity of NK cells against pancreatic cancer PANC1 cells in vitro,and explore its potential mechanism.Methods Peripheral blood mononuclear cells of healthy donors were isolated,NK cells were induced with medium contained with IL-2 and different concentrations of hyperoside (0.3,1.6,8,40 and 200 μg/ml) for 12 days.Cell viability was observed by trypan blue staining.Phenotype and perforin,granzyme B expression of NK cells were detected by flow cytometry.Killing activity of NK cells against PANC1 cells were analyzed with lactate dehydrogenase (LDH) releasing method.Results The proportion of NK cells in control group and experimental group treated with different concentration of hyperoside both reached about 80％,respectively.The proliferation of CDs-CD56 + NK cells treated by hyperoside at 0.3,1.6 and 8 μg/ml was (93.76 ±8.77),(106.67 ± 12.35) and (118.50 ± 11.51) times,respectively,which were significantly higher than (73.70 ± 9.43) times of the control group.The expressions of perforin in NK cells treated with hyperoside at 1.6,8 and 40 μg/ml were significantly higher than those of the control group [(82.34 ± 2.90) ％,(89.15 ±3.54) ％,(81.78 ± 2.81)％ vs (72.93 ± 2.06)％].The expressions of granzyme B in NK cells treated with hyperoside at 1.6 and 8 μg/ml were significantly higher than those of the control group [(87.30 ± 1.70) ％,(92.16 ±3.05)％ vs (82.35 ±2.73)％].The killing activity of NK cells against PANC1 cells treated by hyperoside at 1.6 and 8 μg/ml was significantly higher than those of the control group [(63.18 ± 3.77)％,(65.34 ± 4.97) ％ vs (52.16 ± 5.48) ％].The differences were statistically significant (all P ＜ 0.05).Conclusions Hyperoside could promote the proliferation of NK cells at certain concentrations and maybe enhance the killing effect against pancreatic cancer PANC1 cells through up-regulating the expression of perforin and granzyme B in NK cells.
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Objective To investigate the utility value of monoexponential and biexponential DWI in the differential diagnosis between benign and malignant liver neoplasms.Methods Seventy three patients with pathologically or clinically confirmed liver mass,were analyzed retrospectively and categorized into benign and malignant groups between January 2013 and October 2013.Malignant group included 46 patients with 53 lesions,while 27 patients in benign group had 35 lesions.All patients underwent MR examinations on 3.0T system (GE 750).Conventional MR T1WI,T2WI,DWI(b=0,800 s/mm2) (to obtain ADC with monoexponential modeling),multi-b value DWI(b=0,20 50,100,200,400,600,800 and 1 200 s/mm2) (to obtain Slow-ADC,Fast-ADC,f with biexponential modeling) and dynamic enhancement were performed.The ADC,Slow-ADC,Fast-ADC and f mean values of benign and malignant liver neoplasms were measured and analyzed by using independent samples t test.Diagnostic efficacy of these parameters in malignant group was evaluated by using receiver operating characteristic curve,with histopathologic findings as the gold standard.Results ADC,Slow-ADC,Fast-ADC and f of malignant group were lower than those of benign group [ADC:(1.79±0.35)× 10-3 mm2/s vs (1.16±0.36) × 10-3 mm2/s; Slow-ADC:(1.67±0.25) × 10-3 mm2/s vs(0.94±0.22)×10-3mm2/s; Fast-ADC(72.40±23.70)×10-3mm2/s vs(34.62±17.43)×10-3mm2/s; and f:(33.59± 11.77)％ vs (22.28±8.97)％ in benign and malignant groups,respectively).Significant inter-group difference was observed in ADC,Fast-ADC,Slow-ADC and f (t=0.89,14.77,8.96 and 5.47,respectively and P＜0.05).The areas under the ROC curve (AUC) of ADC,Slow-ADC,Fast-ADC and fwere 0.938,0.974,0.895 and 0.789,respectively.The sensitivity and specificity of ADC,Slow-ADC,Fast-ADC and fwere 90.6％ (48/53),96.2％ (51/53),90.6％ (48/53) and 90.6％ (48/53) and 85.7％ (30/35),91.4％ (32/35),82.9％ (29/35) and 57.1％ (20/35)respectively for differentiating benign from malignant hepatic lesions.Conclusion ADC obtained with mono-exponential modeling and Fast-ADC,Slow-ADC,f obtained with biexponential modeling are useful parameters in distinguishing benign and malignant hepatic lesions,among which slow-ADC demonstrates the highest diagnostic efficacy.
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Objective To investigate the in vitro effects of quercitrin on the proliferation and the cytotoxicity of human γδT cells.Methods Peripheral blood mononuclear cells (PBMCs) were isolated from healthy subjects and cultured with isopentenyl pyrophosphate and IL -2 to induce human γδT cells.The hu-manγδT cells were cultured with quercitrin at various concentrations for 48 hours.CCK-8 kits were used to analyze the in vitro proliferation and cytotoxic activities of γδT cells.Flow cytometry was performed to meas-ure the expression of granzyme B and perforin in γδT cells.The expression of p-ERK, p-Akt and Bcl-2 at protein level were detected by Western blot .Results The percentage of human γδT cells in PBMCs was in-creased from (2.96±1.83)%to (88.94±2.36)%after 10 days of culture.The quercitrin at concentrations of 10 to 80 μg/ml could promote the growth of γδT cells and up-regulate the expression of granzyme B , per-forin, p-ERK, p-Akt and Bcl-2 in a dose dependent manner .The cytolytic activities of γδT cells against co-lonic carcinoma cells ( HCT116 ) were enhanced by quercitrin .Conclusion Quercitrin could promote the proliferation of γδT cells and enhance the expression of granzyme B and perforin at certain concentrations in vitro.ERK1/2 and Akt signal transduction systems might be involved in the process .