Microbial Diversity during Fermentation of Sweet Paste, a Chinese Traditional Seasoning, Using PCR-Denaturing Gradient Gel Electrophoresis

( Ping Mao ),( Yuanliang Hu ),( Tingting Liao ),( Zhaoting Wang ),( Shumiao Zhao ),( Yunxiang Liang ),( Yongmei Hu ) 한국미생물 · 생명공학회 2017 Journal of microbiology and biotechnology Vol.27 No.4

The aim of this study was to elucidate the changes in the microbial community and biochemical properties of a traditional sweet paste during fermentation. PCR-denaturing gradient gel electrophoresis (DGGE) analysis showed that Aspergillus oryzae was the predominant species in the koji (the fungal mixture), and the majority of the fungi isolated belonged to two Zygosaccharomyces species in the mash. The bacterial DGGE profiles revealed the presence of Bacillus subtilis during fermentation, and Lactobacillus acidipiscis, Lactobacillus pubuzihii, Lactobacillus sp., Staphylococcus kloosi, and several uncultured bacteria were also detected in the mash after 14 days of main fermentation. Additionally, during main fermentation, amino-type nitrogen and total acid increased gradually to a maximum of 6.77 ± 0.25 g/kg and 19.10 ± 0.58 g/kg (30 days) respectively, and the concentration of reducing sugar increased to 337.41 ± 3.99 g/kg (7 days). The 180-day fermented sweet paste contained 261.46 ± 19.49 g/kg reducing sugar and its pH value remained at around 4.65. This study has used the PCR-DGGE technique to demonstrate the microbial community (including bacteria and fungi) in sweet paste and provides useful information (biochemical properties) about the assessment of the quality of sweet paste throughout fermentation.

Autophagy inhibition contributes to epigallocatechin-3-gallate-mediated apoptosis in papillary thyroid cancer cells

Bu Ling,Zheng Tingting,Mao Chaoming,Fei Wu,Mou Xiao,Xu Chengcheng,Luo Xuan,Lu Qingyan,Dong Liyang,Wang Xuefeng 대한독성 유전단백체 학회 2021 Molecular & cellular toxicology Vol.17 No.4

Background Epigallocatechin-3-gallate is a natural polyphenolic compound that induces apoptosis in papillary thyroid cancer cells. However, its underlying molecular mechanism was not completely clarified. Objectives The present study demonstrated the role of apoptosis and autophagy in EGCG-treated papillary thyroid cancer cells and the relationship between these processes. Results EGCG significantly suppressed the viability of TPC-1 papillary thyroid cancer cells at an IC50 of 17.2 μM. EGCG induced TPC-1 cell apoptosis and cell cycle arrest at S phase and downregulated the protein expression of cyclin A and cyclin-dependent kinase-2. EGCG decreased reactive oxygen species levels, upregulated Bax expression, downregulated Bcl-2 expression and increased cytochrome C levels in the cytosol. Treatment with EGCG also increased the levels of cleaved caspase 3, cleaved caspase 9 and cleaved poly(ADP-ribose) polymerase. EGCG induced an autophagic response via the upregulation of the autophagy-related protein LC3-II and suppression of the AKT/mTOR signalling pathway. Autophagy inhibition further enhanced EGCG-induced cell apoptosis and ROS suppression, which indicated that autophagy played a cytoprotective role in EGCG-treated TPC-1 cells. Conclusion Taken together, these results demonstrated that autophagy inhibition was beneficial to EGCG–mediated apoptosis in papillary thyroid cancer cells. Background Epigallocatechin-3-gallate is a natural polyphenolic compound that induces apoptosis in papillary thyroid cancer cells. However, its underlying molecular mechanism was not completely clarified. Objectives The present study demonstrated the role of apoptosis and autophagy in EGCG-treated papillary thyroid cancer cells and the relationship between these processes. Results EGCG significantly suppressed the viability of TPC-1 papillary thyroid cancer cells at an IC50 of 17.2 μM. EGCG induced TPC-1 cell apoptosis and cell cycle arrest at S phase and downregulated the protein expression of cyclin A and cyclin-dependent kinase-2. EGCG decreased reactive oxygen species levels, upregulated Bax expression, downregulated Bcl-2 expression and increased cytochrome C levels in the cytosol. Treatment with EGCG also increased the levels of cleaved caspase 3, cleaved caspase 9 and cleaved poly(ADP-ribose) polymerase. EGCG induced an autophagic response via the upregulation of the autophagy-related protein LC3-II and suppression of the AKT/mTOR signalling pathway. Autophagy inhibition further enhanced EGCG-induced cell apoptosis and ROS suppression, which indicated that autophagy played a cytoprotective role in EGCG-treated TPC-1 cells. Conclusion Taken together, these results demonstrated that autophagy inhibition was beneficial to EGCG–mediated apoptosis in papillary thyroid cancer cells.

Study of compensatory growth based on different nutrition conditions of Bombyx mori

Dai Minli,Feng Piao,Mao Tingting,Gu Haoyi,Bian Dandan,Sun Haina,Li Fanchi,Wei Jing,Li Bing 한국응용곤충학회 2022 Journal of Asia-Pacific Entomology Vol.25 No.3

Organisms achieve compensatory growth after a period of nutrient restriction followed by recovering the nutrient status. However, the underlying mechanisms associated with such growth acceleration remain unclear. The silkworm Bombyx mori is a lepidopteran model insect. This study aimed to investigate the physiological characteristics and the underlying mechanisms in B. mori fed on mulberry leaves (MG), artificial diet (AG), and artificial diet + mulberry leaves (AG-MG), respectively. Silkworms in AG-MG which fed on artificial diet from 1st to 3rd instars followed by feeding on mulberry leaves from 4th to 5th instars exhibited a higher weight gain rate than that in MG and AG, indicating that compensatory growth occurred as a result of the switch in the silkworm food regime. Trypsin and lipase activities of silkworms in AG-MG were shown to be up-regulated at 72 h after changing food. Digital gene expression profiling (DGE) analysis revealed that genes related to metabolism and development in silkworm midguts were differentially expressed. The quantitative real-time PCR (qRT-PCR) re sults showed that the expression levels of IIS/PI3K-AKT pathway genes including INR, IRS, AKT, PI3K60 and PI3K110 of silkworms in AG were down-regulated compared with that in MG at 0 h. Whereas AKT, PI3K and PI3K60 of silkworms were significantly increased by 1.68-, 1.49-, 1.67-fold, respectively, at 72 h after switched to mulberry leaves than the same instar fed on artificial diet. PDK’s expression of silkworms in AG was higher than that in other two groups at each timepoint. Compared with MG and AG, PTEN and IRS were down-regulated at 48 and 72 h in AG-MG. Collectively, these results indicate that compensatory growth in B. mori is regulated by IIS/ PI3K-AKT signaling pathway.

Modeling and Virtual Screening of Antisense Peptides Targeting the Divergent Region of Tumor-Associated MT1-MMP Protein

Bowen Tan,Yijie Zhou,Zhilei Song,Yinxuan Peng,Fang Wu,Yue Kang,Xiaomin Liu,Li Zeng,Tingting Huang,Zongying Liu,Lili Xiong,Zhiyun Guo,Jian Cui,Canquan Mao 대한화학회 2015 Bulletin of the Korean Chemical Society Vol.36 No.9

Membrane type-1 matrix metalloproteinase (MT1-MMP; also known as MMP14) is a key enzyme involved in tumor invasion and metastasis, and is a potential target for drug discovery for cancer therapy. However, till now there is no MT1-MMP- or MMP-based anticancer drugs in the market mainly because of the high conservation of the MMP family and also because there is no elucidated crystal structure for the mature MT1-MMP. The modeling of the three-dimensional structure of mature MT1-MMP and the finding of MT1-MMP targeted peptides by virtual screening are highly desired. In this study, the three-dimensional structure of mature MT1-MMP is constructed by homology and de novo modeling and later rationalized and optimized by molecular dynamics simulations. An antisense peptide library was constructed against the divergent sense peptide DEGTEEET in the specific region of MT1-MMP, which was found by multiple alignment of the whole MMP family. The antisense peptide library was virtually screened against the constructed three-dimensional model of MT1-MMP. The top 20 novel peptides were further studied, which were found well docked with MT1-MMP at the region of DEGTEEET, again confirming their specific binding to MT1-MMP. Preliminary study of one of the top-ranked peptide SFLLSPFV showed that it could inhibit the viabilities of MG63 and MDA-MB-231 tumor cells. We thus not only successfully modeled the three-dimensional structure of mature MT1-MMP but also provided a new way for the finding of peptide candidates targeting MT1-MMP based on antisense peptide library.

LincR-PPP2R5C Promotes Th2 Cell Differentiation Through PPP2R5C/PP2A by Forming an RNA–DNA Triplex in Allergic Asthma

Ji Ningfei,Chen Zhongqi,Wang Zhengxia,Sun Wei,Yuan Qi,Zhang Xijie,Jia Xinyu,Wu Jingjing,Jiang Jingxian,Song Meijuan,Xu Tingting,Liu Yanan,Ma Qiyun,Sun Zhixiao,Bao Yanmin,Zhang Mingshun,Huang Mao 대한천식알레르기학회 2024 Allergy, Asthma & Immunology Research Vol.16 No.1

Purpose: The roles and mechanisms of long noncoding RNAs (lncRNAs) in T helper 2 (Th2) differentiation from allergic asthma are poorly understood. We aimed to explore a novel lncRNA, LincR-protein phosphatase 2 regulatory subunit B' gamma (PPP2R5C), in Th2 differentiation in a mouse model of asthma. Methods: LincR-PPP2R5C from RNA-seq data of CD4+ T cells of asthma-like mice were validated and confirmed by quantitative reverse transcription polymerase chain reaction, northern blotting, nuclear and cytoplasmic separation, and fluorescence in situ hybridization (FISH). Lentiviruses encoding LincR-PPP2R5C or shRNA were used to overexpress or silence LincR-PPP2R5C in CD4+ T cells. The interactions between LincR-PPP2R5C and PPP2R5C were explored with western blotting, chromatin isolation by RNA purification assay, and fluorescence resonance energy transfer. An ovalbumin-induced acute asthma model in knockout (KO) mice (LincR-PPP2R5C KO, CD4 conditional LincR-PPP2R5C KO) was established to explore the roles of LincR-PPP2R5C in Th2 differentiation. Results: LncR-PPP2R5C was significantly higher in CD4+ T cells from asthmatic mice ex vivo and Th2 cells in vitro. The lentivirus encoding LincR-PPP2R5C suppressed Th1 differentiation; in contrast, the short hairpin RNA (shRNA) lentivirus decreased LincR-PPP2R5C and Th2 differentiation. Mechanistically, LincR-PPP2R5C deficiency suppressed the phosphatase activity of the protein phosphatase 2A (PP2A) holocomplex, resulting in a decline in Th2 differentiation. The formation of an RNA-DNA triplex between LincR-PPP2R5C and the PPP2R5C promoter enhanced PPP2R5C expression and activated PP2A. LincR-PPP2R5C KO and CD4 conditional KO decreased Th2 differentiation, airway hyperresponsiveness and inflammatory responses. Conclusions: LincR-PPP2R5C regulated PPP2R5C expression and PP2A activity by forming an RNA-DNA triplex with the PPP2R5C promoter, leading to Th2 polarization in a mouse model of acute asthma. Our data presented the first definitive evidence of lncRNAs in the regulation of Th2 cells in asthma.