Characterization of porcine skeletal a-actin gene promoter: expression specificity and regulatory elements

Yujie Zhang,Xu Zhang,Nini Cheng,Yanping Li,Jinyi Xing 한국유전학회 2015 Genes & Genomics Vol.37 No.7

Identification of essential fragments in the promoter region of genes to drive tissue-specific expression of exogenous genes in the skeletal muscle is obligatory for animal transgenic study. The skeletal a-actin is a major protein of the thin filaments of skeletal muscle fiber. In this study, the specificity and activity of porcine skeletal aactin gene promoter were investigated by approaches of cell transfection and assayed with green fluorescent protein (GFP) and dual luciferase reporter activity, respectively. The results revealed that the obtained 3717 bp fragment of porcine skeletal a-actin gene promoter drives GFP to be expressed uniquely in murine C2C12 cells, and two segments of 2.0–2.3 and 0.06–0.37 kb in the promoter region were essential for porcine skeletal a-actin gene expression. The regulatory elements in the 0.06–0.37 kb fragment were further investigated, and when it is deleted or the CArG box within this fragment was mutated, the promoter activity was reduced to 20 or 43 % (P\0.01), respectively, compared to the 3.04 kb segment, suggesting an important role of CArG box in regulating porcine skeletal a-actin gene transcription. The results may provide reference for creating transgenic pigs to express exogenous genes uniquely in pork.

A transcriptional atlas of the silk gland in Antheraea pernyi revealed by IsoSeq

Duan Jianping,Li Shanshan,Zhang Zhengtian,Yao Lunguang,Yang Xinfeng,Ma Sanyuan,Duan Nini,Wang Jiazhen,Zhu Xuwei,Zhao Ping 한국응용곤충학회 2023 Journal of Asia-Pacific Entomology Vol.26 No.2

Silk fibers spun by the silk gland of Antheraea pernyi have many unique properties and are of great value in genetic improvement and non-traditional applications. However, the complete transcriptional landscape and accurate genic annotation of the silk gland are yet to be conducted, which limits related studies on this organ. In this study, isoform sequencing revealed the full-length transcriptome of the A. pernyi silk gland, producing 12,572 high-confidence isoforms from 7,658 genes, among which more than 40 % of genes have not yet been annotated in the reference genome. Moreover, approximately 9 % of isoforms are computationally identified as long non-coding RNAs (lncRNAs). Up to 1,492 alternative splicing (AS) and 3,068 alternative polyadenylation (APA) events were revealed within this transcriptome. In addition, 2,569 putative transcription factors (TFs) belonging to 68 different families were first identified in A. pernyi genome, including 871 TFs in silk gland, and some TF families have undergone expansion or contraction. This study significantly improve our knowledge of the genes expressed in the silk gland of A. pernyi and provide a valuable resource for the in-depth study of silk protein synthesis and spinning, genetic improvement, and non-traditional applications in A. pernyi.

Optimization of ultrasound-assisted extraction of indigo and indirubin from Isatis indigotica Fort. and their antioxidant capacities

Guihong Zhao,Tao Li,Xinyun Qu,Nini Zhang,Miao Lu,Jing Wang 한국식품과학회 2017 Food Science and Biotechnology Vol.26 No.5

An effective method for the ultrasound-assisted extraction of indigo and indirubin from Isatis indigotica Fort. was established and their antioxidant activities were investigated. Response surface methodology based on a three-level, four-factor Box–Behnken design was used to optimize the extraction conditions. Analysis of variance showed that the quadratic model was significant for the extraction of indigo and indirubin (112.72% ± 1.65% and 116.42% ± 1.27%, respectively) under the optimal conditions (methanol concentration, 80%; extraction time, 25 min; ratio of solid to liquid, 1:34 g/mL; and extraction temperature, 41 C) and was in good agreement with the predicted value. Moreover, evaluation of the antioxidant activities suggested that indigo and indirubin presented better scavenging effects on 1,1-diphenyl-2-picrylhydrazyl free radical and superoxide radical than the extract and the extract revealed certain antioxidant activities in hydroxyl radical scavenging and reducing power, and indigo and indirubin could be used as natural antioxidants in the food or medicine industry.

Extracellular signal-regulated kinase 5 increases radioresistance of lung cancer cells by enhancing the DNA damage response

Weiwei Jiang,Guanghui Jin,Fangfang Cai,Xiao Chen,Nini Cao,Xiangyu Zhang,Jia Liu,Fei Chen,Feng Wang,Wei Dong,Hongqin Zhuang,Zichun Hua 생화학분자생물학회 2019 Experimental and molecular medicine Vol.51 No.-

Radiotherapy is a frequent mode of cancer treatment, although the development of radioresistance limits its effectiveness. Extensive investigations indicate the diversity of the mechanisms underlying radioresistance. Here, we aimed to explore the effects of extracellular signal-regulated kinase 5 (ERK5) on lung cancer radioresistance and the associated mechanisms. Our data showed that ERK5 is activated during solid lung cancer development, and ectopic expression of ERK5 promoted cell proliferation and G2/M cell cycle transition. In addition, we found that ERK5 is a potential regulator of radiosensitivity in lung cancer cells. Mechanistic investigations revealed that ERK5 could trigger IR-induced activation of Chk1, which has been implicated in DNA repair and cell cycle arrest in response to DNA double-strand breaks (DSBs). Subsequently, ERK5 knockdown or pharmacological inhibition selectively inhibited colony formation of lung cancer cells and enhanced IR-induced G2/M arrest and apoptosis. In vivo, ERK5 knockdown strongly radiosensitized A549 and LLC tumor xenografts to inhibition, with a higher apoptotic response and reduced tumor neovascularization. Taken together, our data indicate that ERK5 is a novel potential target for the treatment of lung cancer, and its expression might be used as a biomarker to predict radiosensitivity in NSCLC patients.

Tryptophan Metabolites in Irritable Bowel Syndrome: An Overnight Time-Course Study

Robert L Burr,Haiwei Gu,Kevin Cain,Danijel Djukovic,Xinyu Zhang,Claire Han,Nini Callan,Daniel Raftery,,Margaret Heitkemper 대한소화기 기능성질환∙운동학회 2019 Journal of Neurogastroenterology and Motility (JNM Vol.25 No.4

Background/Aims Patients with irritable bowel syndrome (IBS) often report poor sleep quality. Whether poor sleep is associated with tryptophan (Trp) metabolites is unknown. We compared serum Trp metabolites in women with IBS and healthy controls (HCs) using targeted liquid chromatography mass spectrometry (LC-MS)-based profiling. In IBS only, we explored whether Trp metabolites are associated with IBS symptoms and subjective and objective sleep indices, serum cortisol, plasma adrenocorticotropic hormone (ACTH), and cortisol/ACTH levels. Methods Blood samples were obtained every 80 minutes in 21 HCs and 38 IBS subjects following an anticipation-of-public-speaking stressor during a sleep laboratory protocol. Subjects completed symptom diaries for 28 days. Adjacent values of metabolites were averaged to represent 4 time-periods: awake, early sleep, mid-sleep, and mid-to-late sleep. Thirteen of 20 targeted Trp metabolites were identified. Results Ten of 13 Trp metabolites decreased across the night, while nicotinamide increased in both groups. A MANOVA omnibus test performed after principal component analysis showed a significant difference in these 13 principal component (P = 0.014) between groups. Compared to HCs, nicotinamide levels were higher and indole-3-lactic acid levels lower in the IBS group. Melatonin and indole-3-acetic acid levels were associated with several subjective/objective sleep measures; decreased stool consistency/frequency and abdominal pain were positively associated with melatonin and serotonin in the IBS group. The kynurenine and kynurenic acid were associated with ACTH (positively) and cortisol/ACTH (negatively). Conclusions Nighttime Trp metabolites may provide clues to poor sleep and stress with IBS. Further study of the mechanism of metabolite action is warranted.