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peng Hua,Gao Jiao,song Xia 한국식물생명공학회 2022 Plant biotechnology reports Vol.16 No.5
Chloroplast development together with chlorophyll biosynthesis plays an important role in plant survival and growth. Here, EMS-induced chlorophyll-deficient mutants (wtg, wts, and wta) were obtained from the progeny of Mt128 (MT), generated from the maize inbred line DP128 (WT). Three types of leaf color were segregated from the self-fertilized progenies of MT, including green, narrow-white striped and albino and named wtg, wts, and wta, respectively. We found that the Chl and carotenoid contents were significantly changed in wtg, wts, and wta, especially wta, compared with WT. In addition, leaf transcriptome sequencing of wtg, wts, wta, and WT were also carried out. The DEGs were identified as being related to chloroplast development and photosynthesis. By performing pairwise comparisons of each group, a total of 3685 (1181 upregulated, 2504 downregulated), 2624 (1028 upregulated, 1596 downregulated) and 6693 (3018 upregulated, 3675 downregulated) genes were found to be differentially regulated in wtg, wts, and wta, respectively, compared with WT. Of those, 75 comodulated DEGs with great alterations were identified among the comparison groups. More interestingly, most of the DEGs were found to be significantly involved in photosynthesis, including PSI, PSII, the cytochrome b6f complex, and F-type ATPase. GO enrichment of DEGs revealed that most of DEGs were involved in chloroplast development, chlorophyll biosynthesis, or photosynthesis. KEGG enrichment analysis indicated that the five significantly enriched pathways for comodulated DEGs in all EMS-induced mutant genotypes were as follows: benzoxazinoid biosynthesis, photosynthesis, starch and sucrose metabolism, phenylpropanoid biosynthesis and the photosynthesis–antenna proteins pathway. The results showed that EMS induction is a valid approach for generating mutants for crop breeding research.
Song, Chuan-Qing,Zhang, Jun-Hui,Shi, Jia-Chen,Cao, Xiao-Qin,Song, Chun-Hua,Hassan, Adil,Wang, Peng,Dai, Li-Ping,Zhang, Jian-Ying,Wang, Kai-Juan Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.2
Polymorphisms in miRNA binding sites have been shown to affect miRNA binding to target genes, resulting in differential mRNA and protein expression and susceptibility to common diseases. Our purpose was to predict SNPs (single nucleotide polymorphisms) within miRNA binding sites of inflammatory genes in relation to gastric cancer. A complete list of SNPs in the 3'UTR regions of all inflammatory genes associated with gastric cancer was obtained from Pubmed. miRNA target prediction databases (MirSNP, Targetscan Human 6.2, PolymiRTS 3.0, miRNASNP 2.0, and Patrocles) were used to predict miRNA target sites. There were 99 SNPs with MAF>0.05 within the miRNA binding sites of 41 genes among 72 inflammation-related genes associated with gastric cancer. NF-${\kappa}B$ and JAK-STAT are the two most important signaling pathways. 47 SNPs of 25 genes with 95 miRNAs were predicted. CCL2 and IL1F5 were found to be the shared target genes of hsa-miRNA-624-3p. Bioinformatic methods could identify a set of SNPs within miRNA binding sites of inflammatory genes, and provide data and direction for subsequent functional verification research.
Blocking Bcl-2 Leads to Autophagy Activation and Cell Death of the HEPG2 Liver Cancer Cell Line
Du, Peng,Cao, Hua,Wu, Hao-Rong,Zhu, Bao-Song,Wang, Hao-Wei,Gu, Chun-Wei,Xing, Chun-Gen,Chen, Wei Asian Pacific Journal of Cancer Prevention 2013 Asian Pacific journal of cancer prevention Vol.14 No.10
Background: Apoptosis may be induced after Bcl-2 expression is inhibited in proliferative cancer cells. This study focused on the effect of autophagy activation by ABT737 on anti-tumor effects of epirubicin. Methods: Cytotoxic effects of ABT737 on the HepG2 liver cancer cell line were assessed by MTT assay and cell apoptosis through flow cytometry. Mitochondrial membrane potential was measured by fluorescence microscopy. Monodansylcadaverin (MDC) staining was used to detect activation of autophagy. Expression of p53, p62, LC3, and Beclin1, apoptotic or autophagy related proteins, was detected by Western blotting. Results: ABT737 and epirubicin induced growth inhibition in HepG2 cells in a dose- and time-dependent manner. Both ABT737 and epirubicin alone could induce cell apoptosis with a reduction in mitochondrial membrane potential as well as increased apoptotic protein expression. Further increase of apoptosis was detected when HepG2 cells were co-treated with ABT373 and epirubicin. Furthermore, our results demonstrated that ABT373 or epirubicin ccould activate cell autophagy with elevated autophagosome formation, increased expression of autophagy related proteins and LC3 fluorescent puncta. Conclusions: ABT737 influences cancer cells through both apoptotic and autophagic mechanisms, and ABT737 may enhance the effects of epirubicin on HepG2 cells by activating autophagy and inducing apoptosis.
Molecular organization of the B mating type locus of a Lentinula edodes monokaryon strain SUP2
Bao Da-Peng Chen Ming-Jie,Song Wen-Hua,Song Chun-yan,Zhang Mei-Yan,Chen Xiang,Lin Nan 한국버섯학회 2010 한국버섯학회지 Vol.8 No.4
Lentinula edodes is an important cultivated mushroom in China. The development of Lentinula edodes production promotes more studies on it. In our previous work, degenerate PCR and chromosome walking technologies were used to obtain one pheromone receptor gene and one pheromone precursor gene from Lentinula edodes. In this study, four pairs of specific primers were designed according to the whole genome sequencing of the protoplast monokaryon of Lentinula edodes strain 135, to amplify STE3-like pheromone receptor gene and its flanking conserved genes in the protoplast monokaryon strain SUP2 derived from Lentinula edodes strain Suxiang and 33655bp DNA sequence was obtained. By BlastX search, seven putative genes were identified, and three of them are pheromone receptor encoded genes. Furthermore, near to two pheromone receptor genes, four genes encoding proteins with conserved motifs of pheromone precursors were found. This study firstly reveals the molecular organization of the B mating type locus of Lentinula edodes.
( Wei Wang ),( Hua Sheng Wang ),( Hong Bo Hu ),( Hua Song Peng ),( Xue Hong Zhang ) 한국미생물 · 생명공학회 2015 Journal of microbiology and biotechnology Vol.25 No.5
As a global regulatory gene in Streptomyces, afsR can activate the biosynthesis of many secondary metabolites. The effect of afsR on the biosynthesis of a phenazine metabolite, lomofungin, was studied in Streptomyces lomondensis S015. There was a 2.5-fold increase of lomofungin production in the afsR-overexpressing strain of S. lomondensis S015 N1 compared with the wild-type strain. Meanwhile, the transcription levels of afsR and two important genes involved in the biosynthesis of lomofungin (i.e., phzC and phzE) were significantly upregulated in S. lomondensis S015 N1. The optimization of metal chlorides was investigated to further increase the production of lomofungin in the afsR-overexpressing strain. The addition of different metal chlorides to S. lomondensis S015 N1 cultivations showed that CaCl2, FeCl2, and MnCl2 led to an increase in lomofungin biosynthesis. The optimum concentrations of these metal chlorides were obtained using response surface methodology. CaCl2 (0.04 mM), FeCl2 (0.33 mM), and MnCl2 (0.38 mM) gave a maximum lomofungin production titer of 318.0 ± 10.7 mg/l, which was a 4.1-fold increase compared with that of S. lomondensis S015 N1 without the addition of a metal chloride. This work demonstrates that the biosynthesis of phenazine metabolites can be induced by afsR. The results also indicate that metal chlorides addition might be a simple and useful strategy for improving the production of other phenazine metabolites in Streptomyces.
Identification and Functional Analysis of LsMNPV Anti-apoptosis Genes
Kim, Yu-Sin,Xiao, Hua-Zhong,Du, En-Qi,Cai, Guo-Shuai,Lu, Song-Ya,Qi, Yi-Peng Korean Society for Biochemistry and Molecular Biol 2007 Journal of biochemistry and molecular biology Vol.40 No.4
Three anti-apoptosis genes, Ls-iap2, iap3 and p49 were found in Leucania separata multiple nuclear polyhedrovirus. Amino acid sequence homology of Ls-IAP2 and Ls-IAP3 with Op-IAP2 and Op-IAP3 from Orgyia pseddotsugata MNPV were 20% and 42%, while that of Ls-P49 is 28% with Sl-P49 from Spodoptera littorolis MNPV. Ls-IAP2 contains one baculoviral IAP repeat (BIR) domain followed by a RING domain, while Ls-IAP3 contains two BIRs and a RING. Ls-P49 contains a reactive site loop, predicted cleavage site (KKLD$^{74}{\downarrow}$G) that is different from Sl-P49 (TVID$^{94}{\downarrow}$G). Expressed Ls-iap3 or Ls-p49 under presence of actinomycin D in SF9 cells, DNA ladder assayrevealed that Ls- IAP3 or Ls-P49 could block the apoptosis of SF9 cells induced by actinomycin D. Replication of p35 deficient-mutant Autographa californica MNPV in SF9 cells was also rescued when Ls-iap3 or Ls-p49 was expressed transiently. No anti-apoptotic activity was observed for Ls-IAP2. The results showed that both of Ls-IAP3 and Ls-P49 were functional apoptotic suppressors in SF9 cells.
Feng, Xiao-Dong,Song, Qi,Li, Chuan-Wei,Chen, Jian,Tang, Hua-Mei,Peng, Zhi-Hai,Wang, Xue-Chun Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.21
Background: Structural maintenance of chromosomes 4 (SMC-4) is a chromosomal ATPase which plays an important role in regulate chromosome assembly and segregation. However, the role of SMC-4 in the incidence of malignancies, especially colorectal cancer is still poorly understood. Materials and Methods: We here used quantitative PCR and Western blot analysis to examine SMC-4 mRNA and protein levels in primary colorectal cancer and paired normal colonic mucosa. SMC-4 clinicopathological significance was assessed by immunohistochemical staining in a tissue microarray (TMA) in which 118 cases of primary colorectal cancer were paired with noncancerous tissue. The biological function of SMC-4 knockdown was measured by CCK8 and plate colony formation assays. Fluorescence detection has been used to detect cell cycling and apoptosis. Results: SMC-4 expression was significantly higher in colorectal cancer and associated with T stage, N stage, AJCC stage and differentiation. Knockdown of SMC-4 expression significantly suppressed the proliferation of cancer cells and degraded its malignant degree. Conclusions: Our clinical and experimental data suggest that SMC-4 may contribute to the progression of colorectal carcinogenesis. Our study provides a new therapeutic target for colorectal cancer treatment.
Test on the anchoring components of steel shear keys in precast shear walls
Shao-Dong Shen,Peng Pan,Wen-Feng Li,Qi-Song Miao,Run-Hua Gong 국제구조공학회 2019 Smart Structures and Systems, An International Jou Vol.24 No.6
Prefabricated reinforced-concrete shear walls are used extensively in building structures because they are convenient to construct and environmentally sustainable. To make large walls easier to transport, they are divided into smaller segments and then assembled at the construction site using a variety of connection methods. The present paper proposes a precast shear wall assembled using steel shear keys, wherein the shear keys are fixed on the embedded steel plates of adjacent wall segments by combined plug and fillet welding. The anchoring strength of shear keys is known to affect the mechanical properties of the wall segments. Loading tests were therefore performed to observe the behavior of precast shear wall specimens with different anchoring components for shear keys. The specimen with insufficient strength of anchoring components was found to have reduced stiffness and lateral resistance. Conversely, an extremely high anchoring strength led to a short-column effect at the base of the wall segments and low deformation ability. Finally, for practical engineering purposes, a design approach involving the safety coefficient of anchoring components for steel shear keys is suggested.