Effect of Beclin1 overexpression on biological behaviors of SK-MEL-2 human malignant melanoma cells / 中华皮肤科杂志

Objective@#To evaluate the effect of overexpression of the autophagy marker gene Beclin1 on biological behaviors of SK-MEL-2 human malignant melanoma cells.@*Methods@#Western blot analysis was performed to determine the protein expression of Beclin1 in melanoma cell lines A375 and SK-MEL-2. SK-MEL-2 cells with low Beclin1 protein expression were selected as research objects, and divided into 3 groups: blank group receiving no treatment, negative control group transfected with pcDNA.3.1/myc-His (-) A, and experimental group transfected with pcDNA3.1-Beclin1 plasmid. After 2-week culture, cell counting kit-8 (CCK-8) assay was conducted to evaluate the effect of Beclin1 on cell proliferation at 24, 48 and 72 hours, and Transwell assay and wound-healing assay were performed to assess the effect of Beclin1 overexpression on the invasion and migration abilities of SK-MEL-2 cells. Repeated measures analysis of variance and completely randomized analysis of variance were used to analyze differences in indices among groups, and least significant difference (LSD) -t test was used for multiple comparisons.@*Results@#The protein expression of Beclin1 was significantly lower in the SK-MEL-2 cells (0.037 ± 0.010) than in the A375 cells (0.670 ± 0.150, F = 46.62, P<0.05) . The experimental group showed significantly increased protein expression of Beclin1 (0.32 ± 0.04) compared with the negative control group (0.06 ± 0.02, P < 0.05) and blank group (0.07 ± 0.02, P < 0.05) . CCK-8 assay revealed a significant difference in the cell proliferation rate among different groups and different time points (F = 1 077.36, 4 903.04 respectively, both P<0.05) , and there was a significant interaction between the transfection treatment and time (F= 205.20, P<0.05) . Transwell assay showed that the number of SK-MEL-2 cells crossing the chamber per high-power field (× 200) after 24-hour treatment was significantly lower in the experimental group (18.67 ± 1.19) than in the negative control group (87.89 ± 6.05, P<0.05) and blank group (86.78 ± 5.93, P<0.05) . In the wound-healing assay, the cell migration distance was significantly shorter in the experimental group than in the blank group and negative control group at 24 and 48 hours (all P < 0.05) .@*Conclusion@#Beclin1 overexpression can markedly inhibit the proliferation, invasion and migration of SK-MEL-2 cells.

Effect of Beclin1 overexpression on biological behaviors of SK-MEL-2 human malignant melanoma cells / 中华皮肤科杂志

Objective To evaluate the effect of overexpression of the autophagy marker gene Beclin I on biological behaviors of SK-MEL-2 human malignant melanoma cells.Methods Western blot analysis was performed to determine the protein expression of Beclin 1 in melanoma cell lines A375 and SK-MEL-2.SK-MEL-2 cells with low Beclin1 protein expression were selected as research objects,and divided into 3 groups:blank group receiving no treatment,negative control group transfected with pcDNA.3.1/myc-His (-) A,and experimental group transfected with pcDNA3.1-Beclin1 plasmid.After 2-week culture,cell counting kit-8 (CCK-8) assay was conducted to evaluate the effect of Beclin1 on cell proliferation at 24,48 and 72 hours,and Transwell assay and wound-healing assay were performed to assess the effect of Beclin 1 overexpression on the invasion and migration abilities of SK-MEL-2 cells.Repeated measures analysis of variance and completely randomized analysis of variance were used to analyze differences in indices among groups,and least significant difference (LSD)-t test was used for multiple comparisons.Results The protein expression of Beclin1 was significantly lower in the SK-MEL-2 cells (0.037 ± 0.010) than in the A375 cells (0.670 ± 0.150,F =46.62,P ＜ 0.05).The experimental group showed significantly increased protein expression of Beclin1 (0.32 ± 0.04) compared with the negative control group (0.06 ± 0.02,P ＜0.05) and blank group (0.07 ± 0.02,P ＜ 0.05).CCK-8 assay revealed a significant difference in the cell proliferation rate among different groups and different time points (F =1 077.36,4 903.04 respectively,both P＜ 0.05),and there was a significant interaction between the transfection treatment and time (F =205.20,P ＜ 0.05).Transwell assay showed that the number of SK-MEL-2 cells crossing the chamber per high-power field (× 200) after 24-hour treatment was significantly lower in the experimental group (18.67 ±1.19) than in the negative control group (87.89 ± 6.05,P＜ 0.05) and blank group (86.78 ± 5.93,P ＜0.05).In the wound-healing assay,the cell migration distance was significantly shorter in the experimental group than in the blank group and negative control group at 24 and 48 hours (all P ＜ 0.05).Conclusion Beclin 1 overexpression can markedly inhibit the proliferation,invasion and migration of SK-MEL-2 cells.

Comparison of the two magnetic resonance deartifact techniques in imaging of different porcelain-fused-to-metal crowns / 华西口腔医学杂志

OBJECTIVE@#This study aimed to compare the porcelain-fused-to-metal (PFM) crown artifact in the magnetic resonance imaging (MRI) of the two magnetic resonance deartifact techniques in studying the application value of the propeller-fast spin-echo T2-weighted sequence (FSE T2WI) in troubleshooting PFM crown artifacts.@*METHODS@#A total of 48 patients with right mandible first molar crown who underwent MRI head examination were chosen as subjects in the study. According to different metal substrates, PFM crowns were divided to three types, namely, nickel-chromium alloy crown, cobalt-chromium alloy crown and titanium crown. The patients received two MRI scan sequences, that is, FSE T2WI and propeller-FSE T2WI sequences. The MRI artifacts areas in two sequences were measured.@*RESULTS@#The difference between FSE T2WI and propeller-FSE T2WI sequences in three kinds of PFM crown was significant (P<0.05).@*CONCLUSIONS@#Propeller-FSE T2WI sequence technique can effectively reduce the metal artifacts of various PFM crowns.

Loss of IκB kinase β promotes myofibroblast transformation and senescence through activation of the ROS-TGFβ autocrine loop

Using forward and reverse genetics and global gene expression analyses, we explored the crosstalk between the IκB kinase β (IKKβ) and the transforming growth factor β (TGFβ) signaling pathways. We show that in vitro ablation of Ikkβ in fibroblasts led to progressive ROS accumulation and TGFβ activation, and ultimately accelerated cell migration, fibroblast-myofibroblast transformation and senescence. Mechanistically, the basal IKKβ activity was required for anti-oxidant gene expression and redox homeostasis. Lacking this activity, IKKβ-null cells showed ROS accumulation and activation of stress-sensitive transcription factor AP-1/c-Jun. AP-1/c-Jun activation led to up-regulation of the Tgfβ2 promoter, which in turn further potentiated intracellular ROS through the induction of NADPH oxidase (NOX). These data suggest that by blocking the autocrine amplification of a ROS-TGFβ loop IKKβ plays a crucial role in the prevention of fibroblast-myofibroblast transformation and senescence.

Diffusion weighted MRI helps evaluate angiogenesis and vascular endothelial growth factor expression in prostate cancer / 中华男科学杂志

<p><b>OBJECTIVE</b>To study whether diffusion weighted imaging (DWI) and apparent diffusion coefficient (ADC) can reflect angiogenesis and the expression of the vascular endothelial growth factor (VEGF) by analyzing the correlation between the features of DWI and angiogenesis in prostate cancer (PCa).</p><p><b>METHODS</b>We studied the clinical and pathological data of 38 patients with histologically proven PCa, who were examined in the supine position with a 1.5T superconductive magnetic scanner (Siemens Sonata) with a pelvic phased array multi-coil. DWI was obtained by echo planar imaging (EPI) sequence. Another 33 cases of benign prostate hyperplasia (BPH) and 15 healthy volunteers were detected for the ADC value in the PCa and BPH tissues and the peripheral zone (PZ). All the PCa samples were examined for microvascular density (MVD) and VEGF.</p><p><b>RESULTS</b>The ADC values of PCa, BPH and PZ were (49.32 +/- 12.68) x 10(-5) mm2/s, (86.73 +/- 26.75) x 10(-5) mm2/s and (126.25 +/- 27.21) x 10(-5) mm2/s, the former lower than the latter two (P < 0.05). The expressions of MVD and VEGF in PCa were higher than in BPH (P < 0.05). The correlation was negative between the ADC value and MVD of PCa (r = -0.510, P < 0.05) , and positive between the expressions of VEGF and MVD (r = 0.481, P < 0.05). The ADC values of the VEGF-positive and -negative groups were (47.27 +/- 9.55) x 10(-5) mm2/s and (55.06 +/- 11.6) x 10(-5) mm2/s (P < 0.05).</p><p><b>CONCLUSION</b>The ADC value reflects the angiogenesis in differentiated prostate cancer, and DWI therefore helps to evaluate the biological features of PCa in vivo.</p>

Quantitative monomolecular coverage of barium glass by the silane coupling agent gamma-methacryloxypropyltrimethoxysilane / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To study the monomolecular coverage of the silane coupling agent gamma-methacryloxypropyltrimethoxysilane (gamma-MPS) on the barium glass filler surface.</p><p><b>METHODS</b>Kubelka-Munk (K-M) function values of the isolated OH-groups on the barium glass filler surface, which were based on the change of gamma-MPS concentrations, were measured using the quantitative analysis of the diffuse reflectance infrared Fourier transform (DRIFT) spectroscopy. When all of the isolated OH-groups on the filler surface disappeared (where the K-M function values was zero), the monomolecular coverage of the gamma-MPS molecules on the filler surface was indicated by the linear regression analysis.</p><p><b>RESULTS</b>The relationship tallied with negative linear correlation between the K-M function values of the isolated OH-groups on the barium glass surface and the concentrations of gamma-MPS. Each gamma-MPS molecule occupied 0.21 nm(2) when the monomolecular coverage was formed on the barium glass surface.</p><p><b>CONCLUSIONS</b>The result of this study indicated the optimal amount of silane coupling agent on silanated barium glass filler during the production of resin composite.</p>