Characterization of Cucurbita maxima Fruit Metabolomic Profiling and Transcriptome to Reveal Fruit Quality and Ripening Gene Expression Patterns

He-Xun Huang,Ting Yu,Jun-Xing Li,Shu-Ping Qu,왕만만,Ting-Quan Wu,Yu-Juan Zhong 한국식물학회 2019 Journal of Plant Biology Vol.62 No.3

Pumpkin (Cucurbita maxima) fruit is the importantdietary source of carotenoid and is known for the goodflavour and texture due to the accumulation of sugar andstarch. However, lack of transcriptional information hindersour understanding of the molecular mechanisms underlyingfruit quality attributes and nutrition in C. maxima. To provideinsight into transcriptional regulation of fruit quality formationof C. maxima, quality analysis and high-throughput RNAsequencing of fruits at different developing stages werecharacterized. The quality analyses consist of dry mattervalues, percent soluble solids, carotenoid contents, and starchand sugar contents in seven stages of fruit development. Fruit transcriptome of C. maxima at five stages throughoutdevelopment was assembled to elucidate the molecularregulation of fruit development. Almost 18 billion nucleotidebases were sequenced in total, and 48,471 unigenes weredetected. A total of 32,397 (66.8%) unigenes were identifiedto be differentially expressed. We found there was a correlationbetween ripening-associated transcripts and metabolites andthe functions of regulating genes. KEGG analysis showedthere are multiple transcripts enriched in starch, sugar, carotenoid,plant hormone signal transduction and pectin pathways andseveral pathways regulating quality formation were identified. Candidate genes involving in sugar, starch, pectin, fruitsoftening and carotenoid metabolism in fruit were firstlyidentified for the species of C. maxima. Combining the sugar, starch and carotenoid accumulating patterns duringfruit development, a series of possible rate limiting geneswere identified. These findings will provide valuable informationfor further studies regarding fruit quality and development.

Investigation of factors affecting the thermal stability of silica-based shear thickening fluids

Wang Wenjian,He Shuai,Dong Ziwen,Huang Wei,Li Xuke,Chen Xun,Chen Peng 한국유변학회 2024 Korea-Australia rheology journal Vol.36 No.3

Recent experimental observation in thermal-induced gelation of silica-based shear thickening fluids (STFs) has necessitated the efforts to address the thermal instability of the STFs. The shear thickening behavior of hydrophilic fumed silica/PEG disappeared after thermal treatment. Hydrophobic surface is demonstrated to enhance thermal stability of the fumed silica/ PEG system under the same thermal treatment. Dynamic temperature sweep experiments reveal a temperature-induced sol–gel transition of the hydrophilic silica/PEG. Meanwhile, hydrophobic silica/PEG does not undergo such gelation and still displays viscosity thickening after thermal treatment. This gelation process is found to be concentration-dependent and irreversible, with enhanced gelation at higher specific surface areas of hydrophilic silica particles. The small-angle X-ray scattering (SAXS) studies suggest that this sol–gel transition is likely attributed to the reduction of solvation layer which could be highly influenced by particle surface hydrophilicity. Comprehensive experiments, including Soxhlet extraction and rheology, confirm that the resulting stable three-dimensional structure is not formed by chemical crosslinks but behaves as a physical network. It is demonstrated how tailoring particle–medium interactions through controlling particle surface characteristics can be used to improve thermal stability of silica-based STFs.

Soluble Expression of Recombinant Human Smp30 for Detecting Serum Smp30 Antibody Levels in Hepatocellular Carcinoma Patients

Zhang, Sheng-Chang,Huang, Peng,Zhao, Yong-Xiang,Liu, Shu-Yan,He, Shu-Jia,Xie, Xiao-Xun,Luo, Gou-Rong,Zhou, Su-Fang Asian Pacific Journal of Cancer Prevention 2013 Asian Pacific journal of cancer prevention Vol.14 No.4

Senescence marker protein 30 (SMP30), a hepatocellular carcinoma (HCC) associated antigen, was earlier shown by our research group to be highly expressed in HCC paracancerous tissues, but have low levels in HCC tissues. In order to detect anti-SMP30 antibody in serum of HCC patients, we established pET30a-SMP30 and pColdIII-SMP30 expression systems in Escherichia coli. However, the expression product was mainly in the form of inclusion bodies. In this research, we used several combinations of chaperones, four molecular chaperone plasmids with pET30a-SMP30 and five molecular chaperone plasmids with pColdIII-SMP30 to increase the amount of soluble protein. Results showed that co-expression of HIS-SMP30 with pTf16, combined with the addition of osmosis-regulator, and a two-step expression resulted in the highest enhancement of solubility. A total of 175 cases of HCC serum were studied by ELISA to detect anti-SMP30 antibody with recombinant SMP30 protein. Some 22 were positive and x2 two-sided tests all showed P>0.05, although it remained unclear whether there was a relationship between positive cases and clinical diagnostic data.

Dryocrassin ABBA Induces Apoptosis in Human Hepatocellular Carcinoma HepG2 Cells Through a Caspase-Dependent Mitochondrial Pathway

Jin, Zhe,Wang, Wen-Fei,Huang, Jian-Ping,Wang, He-Meng,Ju, Han-Xun,Chang, Ying Asian Pacific Journal of Cancer Prevention 2016 Asian Pacific journal of cancer prevention Vol.17 No.4

Background: Biological and pharmacological activities of dryocrassin ABBA, a phloroglucinol derivative extracted from Dryopteris crassirhizoma, have attracted attention. In this study, the apoptotic effect of dryocrassin ABBA on human hepatocellular carcinoma HepG2 cells was investigated. Materials and Methods: We tested the effects of dryocrassin ABBA on HepG2 in vitro by MTT, flow cytometry, real-time PCR, and Western blotting. KM male mice were used to detect the effect of dryocrassin ABBA on H22 cells in vivo. Results: Dryocrassin ABBA inhibited the growth of HepG2 cells in a concentration-dependent manner. After treatment with 25, 50, and $75{\mu}g/mL$ dryocrassin ABBA, the cell viability was 68%, 60% and 49%, respectively. Dryocrassin ABBA was able to induce apoptosis, measured by propidium iodide (PI)/annexin V-FITC double staining. The results of real-time PCR and Western ting showed that dryocrassin ABBA up-regulated p53 and Bax expression and inhibited Bcl-2 expression which led to an activation of caspase-3 and caspase-7 in the cytosol, and then induction of cell apoptosis. In vivo experiments also showed that dryocrassin ABBA treatment significantly suppressed tumor growth, without major side effects. Conclusions: Overall, these findings provide evidence that dryocrassin ABBA may induce apoptosis in human hepatocellular carcinoma cells through a caspase-mediated mitochondrial pathway.

Knockdown of GCF2/LRRFIP1 by RNAi Causes Cell Growth Inhibition and Increased Apoptosis in Human Hepatoma HepG2 Cells

Li, Jing-Ping,Cao, Nai-Xia,Jiang, Ri-Ting,He, Shao-Jian,Huang, Tian-Ming,Wu, Bo,Chen, De-Feng,Ma, Ping,Chen, Li,Zhou, Su-Fang,Xie, Xiao-Xun,Luo, Guo-Rong Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.6

Background: GC-binding factor 2 (GCF2) is a transcriptional regulator that represses transcriptional activity of the epidermal growth factor receptor (EGFR) by binding to a specific GC-rich sequence in the EGFR gene promoter. In addition to this function, GCF2 has also been identified as a tumor-associated antigen and regarded as a potentially valuable serum biomarker for early human hepatocellular carcinoma (HCC) diagnosis. GCF2 is high expressed in most HCC tissues and cell lines including HepG2. This study focused on the influence of GCF2 on cell proliferation and apoptosis in HepG2 cells. Materials and Methods: GCF2 expression at both mRNA and protein levels in HepG2 cells was detected with reverse transcription (RT) PCR and Western blotting, respectively. RNA interference (RNAi) technology was used to knock down GCF2 mRNA and protein expression. Afterwards, cell viability was analyzed with a Cell Counting Kit-8 (CCK-8), and cell apoptosis and caspase 3 activity by flow cytometry and with a Caspase 3 Activity Kit, respectively. Results: Specific down-regulation of GCF2 expression caused cell growth inhibition, and increased apoptosis and caspase 3 activity in HepG2 cells. Conclusions: These primary results suggest that GCF2 may influence cell proliferation and apoptosis in HepG2 cells, and also provides a molecular basis for further investigation into the possible mechanism at proliferation and apoptosis in HCC.