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Intramuscular fat (IMF) is an important trait influencing meat quality, and intramuscular stromal-vascular cell (MSVC) differentiation is a key factor affecting IMF deposition. Quantitative real-time PCR (qPCR) is often used to screen the differentially expressed genes during differentiation of MSVCs, where proper reference genes are essential. In this study, we assessed 31 of previously reported reference genes for their expression suitability in porcine MSVCs derived form longissimus dorsi with qPCR. The expression stability of these genes was evaluated using NormFinder, geNorm and BestKeeper algorithms. NormFinder and geNorm uncovered ACTB, ALDOA and RPS18 as the most three stable genes. BestKeeper identified RPL13A, SSU72 and DAK as the most three stable genes. GAPDH was found to be the least stable gene by all of the three software packages, indicating it is not an appropriate reference gene in qPCR assay. These results might be helpful for further studies in pigs that explore the molecular mechanism underlying IMF deposition. ImplicationsThis study evaluated the expression stability of a set of well-known reference genes in porcine intramuscular stromal-vascular cells (MSVCs), whose adipogenic differentiation is critical for intramuscular fat deposition and pork quality. ACTB, ALDOA and RPS18 were identified to be most stably expressed throughout adipogenic differentiation, and suitable to be used for normalization in quantitative realtime PCR assay, whereas GAPDH was the last choice. This study has provided a valuable data for our further work to explore the molecular mechanism underlying porcine MSVCs adipogenic differentiation and intramuscular fat deposition.

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Studies on the voltage distribution on the top electrode are important to improve the radio frequency (RF) heating uniformity of samples. The goal of this study was to evaluate the top electrode voltage distribution uniformity in RF systems. The voltages of five representative points on the top electrode were measured by a measuring device with 3 kg soybeans, and validated by the heating patterns using 0.9 kg soybeans located at five positions of the RF cavity. The results showed that the length effect of the copper straps used for connecting the measuring points and measuring device should be taken into account by ignoring little effects of the strap shapes. The final voltages away from the feed strip were higher than those near the feed strip and the deviations between the minimum and the maximum values among the five voltages were 694.3 and 440.9 V, corresponding to the highest voltage difference of 11.99 and 7.12% without and with load, respectively. The real top electrode voltage distribution could be potentially used in computer simulations to achieve the accurate RF heating patterns.