OBJECTIVETo establish an efficient and high resolution 2-DE (two-dimensional electrophoresis) protocol for root tuber proteome analysis of Rehmannia glutinosa.

METHODProteins from root tuber of R. glutinosa were extracted by using five different methods and their productivity and profiles were assessed by means of SDS-PAGE and two-dimensional electrophoresis.

RESULTThe trichloroacetic acid (TCA)-phenol extraction method was found most effective for the extraction with the highest protein yield, the most spots in protein patterns, and the highest resolution of proteins, and the clearest background could be achieved simultaneously. A 1:5 solution of ampholine pH 3-10 and pH 5-8 for a nonlinear gel and the 170 microg of protein loading dosage obtained maps with more protein spots and higher resolution of separation patterns.

CONCLUSIONThis study based on the optimized root tuber proteome preparation and the 2-DE protocol gets a high resolution and reproducibility 2-DE image, which will be expected to have excellent applications in proteomics studies of R. glutinosa tuber root.

Electrophoresis, Gel, Two-Dimensional ; methods ; Plant Proteins ; analysis ; chemistry ; Plant Roots ; chemistry ; Proteome ; chemistry ; isolation & purification ; Rehmannia ; chemistry

Objective: To investigate the effect of the multi-material artifact reduction (MMAR) algorithm of spectral CT in reducing the beam hardening artifacts in dental restoration material. Methods: Three-unit fixed bridge restorations were fabricated on the first to third molars in pig jaw. Gold alloy, zirconia, cobalt chromium alloy, nickel chromium alloy and pure titanium were used as materials for these fixed bridges. After restoration delivery, the pig jaw was scaned using energy spectrum CT machines. Images in regular 120 kVp scan mode were used as conventional group, and reconstructed single-energy horizontal images of 80, 90, 100, 110, 120, 130 and 140 keV in energy spectrum scanning mode were used as energy spectrum group, and reconstructed images applied MMAR technology in energy spectrum scanning mode were used as energy spectrum MMAR group. Each group was scanned 10 times to measure CT value and noise of muscles around dental prosthetic materials and adjacent non-artifact layers. Artifact index was calculated. Two radiologists scored the image quality of each group subjectively. Kruskal Wallis rank sum test was used to compare the difference of image noise, artifact index and subjective score among the control group and the best keV condition in the energy spectrum group and the energy spectrum MMAR group. Results: The image noise of energy spectrum group and energy spectrum MMAR group decreased gradually with the increase of single energy level. The artifact index of pure titanium restorations in conventional group, energy spectrum group and energy spectrum MMAR group were 71.0±8.0, 21.4±2.7 and 14.7±2.7 respectively, and these values were significantly lower than those of other materials in the same group (P<0.05). The subjective image quality scores in energy spectrum MMAR group were as follows: 3.0±0.2 for gold alloy, 4.3±0.5 for zirconia, 3.0±0.4 for cobalt chromium alloy, 3.1±0.4 for cobalt chromium alloy, and 4.6±0.5 for pure titanium. These scores were significantly smaller than those in the conventional group (P<0.05). There was no significant difference in noise between energy spectrum group and energy spectrum MMAR group (P>0.05), and the noise values in energy spectrum group and energy spectrum MMAR group were significantly lower than that in the conventional group (P<0.05). Conclusions Artifacts of pure titanium is minimal. Energy spectrum CT combined with MMAR technology can effectively reduce the artifacts of gold, zirconia, cobalt-chromium alloy, nickel-titanium alloy and pure titanium. This technique can be used as an effective method to remove artifacts of dental prosthesis.