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Pleural effusion (PE) is commonly observed in advanced lung cancer patients. Cell-free total nucleic acid (cfTNA) isolated from cancer patients' plasma has allowed noninvasive tumor genome analyses; however, there are limited studies of detection and characterization of cfTNA in PE. Herein, we included 47 advanced non-small-cell lung cancer patients with PE, who had lung cancer driver mutations tested on tumor tissue specimens either at diagnosis or during disease progression. The supernatant and cell pellet of each PE were evaluated for molecular profiles in parallel on an Ion Torrent next-generation sequencing platform. Somatic mutations were detected in 89.1% supernatant cfTNA, but in only 54.3% of cell pellets. The overall concordance rate between supernatants and formalin-fixed, paraffin-embedded cell pellets at the mutation level was 53.3%. By contrast, 41.7% and 5.0% of somatic alterations were detected in supernatants and cell pellets, respectively. Furthermore, joint analysis of supernatants and cell pellets from PE showed a high concordance (88.3%) of variant detection with their respective tumor tissue specimens. Low-frequency T790M mutations in three cases (0.29%, 0.41%, and 1.56%) were detected in supernatants but not in the matched cell pellets or tumor tissues. In conclusion, pleural effusion-derived cfTNA can effectively be used in clinical practice for molecular analysis by next-generation sequencing, even in cases where corresponding cell pellets or tumor tissues yield insufficient material.

Tribolium castaneum (Herbst) ranks as one of the most prevalent insects in food processing and storage facilities worldwide. Heat treatment has been revisited to disinfest food processing and storage facilities due to increasingly strict regulation on chemicals. The effect of acclimation of T. castaneum larvae to sublethal high temperatures of 36 and 42℃ for 10 h on their heat adaptation was investigated, and transcript-level analysis combinating with real-time PCR (RT–qPCR) was applied for elucidating the heat adaptation mechanism of T. castaneum larvae. Short-term sublethal high temperature acclimation could greatly enhance the thermal adaptability in T. castaneum larvae. In total, 575, 875, and 1017 differentially expressed genes (DEGs) were, respectively, determined in comparisons between the 28 and 36℃ treatments, the 28 and 42℃ treatments, and the 36 and 42℃ treatments. Fifty-three and 96 genes were commonly up- and down-regulated in both the 36 and 42℃ treatments relative to 28℃, respectively. The results of RT-qPCR analysis further confirmed the RNA-seq analysis. The current results are in favor of enhancing the insecticidal effectiveness of extreme high temperature treatment and elucidating the heat adaptation mechanism in T. castaneum larvae.

Tribolium castaneum is one of the most destructive insects infesting cereals worldwide. Heat treatment, as one of the promising alternatives to chemical control, has been widely applied to disinfest empty grain storage spaces and food processing facilities. The influence of acclimation to sublethal high temperatures (36 and 42 °C for 10 h) on the heat tolerance of T. castaneum adults was investigated, and the transcriptome of T. castaneum adults was sequenced using Illumina sequencing technology for exploring the heat tolerance mechanism of T. castaneum adults. Acclimation to sublethal high temperatures significantly enhanced heat tolerance of T. castaneum adults. A total of 12,596 unigene sequences were annotated with T. castaneum reference genome. A total of 280, 1134, and 948 DEGs were respectively identified in the 28A-36A, 28A-42A, and 36A-42A. Seventy-three commonly up-regulated genes and 36 commonly down-regulated genes were confirmed in response to heat tolerance from 28A to 36A, and 28A-42A. The results of RT-qPCR analysis were high significantly consistent with the RNA-seq analysis. These results offer a valuable information for future improving the effectiveness of heat treatment and exploring the evolution mechanism of the heat tolerance of T. castaneum adults.

The influence of packaging materials on infestation by Tribolium castaneum (Herbst) in wheat flour was investigated under laboratory conditions at 27 ± 2and 75 ± 5% relative humidity. The tested packaging materials included vacuum-sealed plastic bags, aluminum foil bags, non-woven cloth bags, kraft paper bags, and plastic compound bags. When T. castaneum adults were released on packaged wheat flour, the degree of infestation varied depending on the packaging materials. The highest infestation was observed in refined wheat flour packaged with plastic compound bags (mean 46 ± 20 insects). For the flour packaged with non-woven cloth bag and kraft paper bags exposed to adults, insect infestation was nil or negligible (mean population < 3 ± 1 insects). For the flour packaged with aluminum foil bags and vacuum-sealed plastic bags exposed to adults, insect infestation was nil. Damage to the packaging materials along the edges or on the upper surfaces was observed in non-woven cloth bags and plastic compound bags. Aluminum foil bags and vacuum-sealed plastic bags offered the best protection against T. castaneum adults.