Identification and comparative analysis of the pearl oyster Pinctada fucata hemocytes microRNAs in response to Vibrio alginolyticus infection

Zhongliang Wang,Bei Wang,Gang Chen,Yishan Lu,Jichang Jian,Zaohe Wu 한국유전학회 2017 Genes & Genomics Vol.39 No.10

MicroRNAs (miRNAs) are a class of noncoding RNA molecules that function as negative regulators of gene expression and play important roles in a wide spectrum of biological processes, including in immune response. However, the physiological regulation function of Pinctada fucata miRNAs, specially their immunomodulation has not been explored yet. Here, two small RNA libraries from hemocytes of P. fucata with or without Vibrio alginolyticus infection were constructed and sequenced using the high-throughput Illumina deep sequencing technology. In total, 11,939,992 and 11,083,327 raw reads, corresponding to 10,993,546 and 9,988,179 clean reads, were respectively obtained in the control and infected libraries. A total of 276 miRNAs, including 225 known miRNAs and 51 putative novel miRNAs, were identified by bioinformatic analysis. By using pairwise comparison between two libraries, 93 miRNAs were found to be significantly differentially expressed, with 42 and 51 miRNAs exhibiting up-regulation and down-regulation, respectively. Thereinto, some known miRNAs were considered to be immune-related. Real-time PCR were implemented for 6 miRNAs co-expressed in the control and infected samples, and agreement was confirmed between the high-throughput sequencing and real-time PCR data. After miRNA targets were predicted, GO and KEGG pathway enrichment analysis were performed, and the results indicated that ten of the differentially expressed miRNAs were involved in immunerelated pathways, and might participate in the host immune response to V. alginolyticus. These results of identification and comparative analysis of miRNAs might deepen our understanding of host-pathogen interactions and immune defense mechanisms in P. fucata.

Expression of VEGF-C/VEGFR-3 in Human Laryngeal Squamous Cell Carcinomas and its Significance for Lymphatic Metastasis

Wang, Zhongliang,Chen, Yao,Li, Xiaofeng,Xu, Li,Ma, Wei,Chang, Lingmei,Ju, Funian Asian Pacific Journal of Cancer Prevention 2012 Asian Pacific journal of cancer prevention Vol.13 No.1

Objectives: Expression of vascular endothelial growth factor C (VEGF-C)and vascular endothelial growth factor feceptor-3 (VEGFR-3) in laryngeal squamous carcinoma and its relationship to lymph node metastasis were investigated. Methods: VEGF-C and VEGFR-3 gene expression in 30 cases of normal laryngeal mucosa tissue (NLM), primary laryngeal carcinoma cell carcinomas (PLC) and cervical lymph nodes (CLN) was examined by reverse transcription polymerase chain reaction (RT-PCR). Protein levels of VEGF-C expression were determined by immunohistochemical staining in 60 cases of PLC. Results: Expression of VEGF-C and VEGFR-3 different among NLM, PLC and CLN in the same patient. In PLC, expression was significantly higher in lymph node positive group than in the lymph node negative group and associated with histological grade of differentiation; Expression of VEGF-C and VEGFR-3 was not linked with age, sex, site or T stage. Conclusions: A close correlation was found between VEGF-C/VEGFR-3 expression and lymph node metastasis in PLC, suggesting a role in metastasis of laryngeal carcinomas.

Regulation of chicken vanin1 gene expression by peroxisome proliferators activated receptor α and miRNA-181a-5p

Wang, Zhongliang,Yu, Jianfeng,Hua, Nan,Li, Jie,Xu, Lu,Yao, Wen,Gu, Zhiliang Asian Australasian Association of Animal Productio 2021 Animal Bioscience Vol.34 No.2

Objective: Vanin1 (VNN1) is a pantetheinase that can catalyze the hydrolysis of pantetheine to produce pantothenic acid and cysteamine. Our previous studies showed that VNN1 is specifically expressed in chicken liver. In this study, we aimed to investigate the roles of peroxisome proliferators activated receptor α (PPARα) and miRNA-181a-5p in regulating VNN1 gene expression in chicken liver. Methods: 5'-RACE was performed to identify the transcription start site of chicken VNN1. JASPAR and TFSEARCH were used to analyze the potential transcription factor binding sites in the promoter region of chicken VNN1 and miRanda was used to search miRNA binding sites in 3' untranslated region (3'UTR) of chicken VNN1. We used a knock-down strategy to manipulate PPARα (or miRNA-181a-5p) expression levels in vitro to further investigate its effect on VNN1 gene transcription. Luciferase reporter assays were used to explore the specific regions of VNN1 targeted by PPARα and miRNA-181a-5p. Results: Sequence analysis of the VNN1 promoter region revealed several transcription factor-binding sites, including hepatocyte nuclear factor 1α (HNF1α), PPARα, and CCAAT/enhancer binding protein α. GW7647 (a specific agonist of PPARα) increased the expression level of VNN1 mRNA in chicken primary hepatocytes, whereas knockdown of PPARα with siRNA increased VNN1 mRNA expression. Moreover, the predicted PPARα-binding site was confirmed to be necessary for PPARα regulation of VNN1 gene expression. In addition, the VNN1 3'UTR contains a sequence that is completely complementary to nucleotides 1 to 7 of miRNA-181a-5p. Overexpression of miR-181a-5p significantly decreased the expression level of VNN1 mRNA. Conclusion: This study demonstrates that PPARα is an important transcriptional activator of VNN1 gene expression and that miRNA-181a-5p acts as a negative regulator of VNN1 expression in chicken hepatocytes.

DNA methyltransferase-3a interacts with p53 and represses p53-mediated gene expression.

Wang, Y Alan,Kamarova, Yeugeniya,Shen, Kate C,Jiang, Zhongliang,Hahn, Myong-Joon,Wang, Yaolin,Brooks, S C Landes Bioscience 2005 Cancer biology & therapy Vol.4 No.10

<P>Genome stability maintenance is regulated by both genetic and epigenetic mechanisms. DNA methylation is the predominant epigenetic mechanism in regulation of gene expression and in suppression of mobile DNA elements from random integration in the genome. The importance of DNA methylation in tumorigenesis has been demonstrated in cancer cells, which harbor global genomic DNA hypomethylation and regional hypermethylation at CpG islands of tumor suppressor genes. DNA methylation is mediated by a class of DNA methyltransferases (Dnmts) involved in de novo methylation of genomic DNA and in the maintenance of DNA methylation patterns during replication. Global genomic DNA demethylation induced by 5-Aza-deoxycytidine activates the p53 signaling pathway and induces apoptosis, suggesting that DNA methylation mediated by Dnmts is associated with p53 signaling in maintaining genome stability. In this report, we show that Dnmt3a interacts with p53 directly and represses p53-mediated transactivation of the p21 gene. It was found that trans-repression by Dnmt3a does not require the methyltransferase activity implying that transcriptional repression does not involve promoter silencing through DNA methylation by Dnmt3a. Finally, the activity of Dnmt3a in vivo was demonstrated when this enzyme was overexpressed in a breast cell line in which Dnmt3a repressed p21 upregulation following DNA damage. The results presented in this study provide new understanding of tumor promotion as mediated by Dnmt3a through its interaction with p53, and suppression of the p53-mediated transcription of tumor suppressor genes. Given that the expression of Dnmts is increased in certain cancers, it is likely that increased Dnmts could block the transactivation function of p53 following its induction by chemotherapeutic drugs resulting in chemoresistance. The use of a DNA methyltransferase inhibitor would therefore restore the p53 tumor suppression function and the utilization of such an inhibitor in combination with DNA damage agents might be an effective therapy for certain cancers.</P>

CRISPR/Cas9-mediated knockout of the Vanin-1 gene in the Leghorn Male Hepatoma cell line and its effects on lipid metabolism

Xu Lu,Wang Zhongliang,Liu Shihao,Wei Zhiheng,Yu Jianfeng,Li Jun,Li Jie,Yao Wen,Gu Zhiliang 아세아·태평양축산학회 2024 Animal Bioscience Vol.37 No.3

Objective: Vanin-1 (VNN1) is a pantetheinase that catalyses the hydrolysis of pantetheine to produce pantothenic acid and cysteamine. Our previous studies have shown that the VNN1 is specifically expressed in chicken liver which negatively regulated by microRNA-122. However, the functions of the VNN1 in lipid metabolism in chicken liver haven’t been elucidated. Methods: First, we detected the VNN1 mRNA expression in 4-week chickens which were fasted 24 hours. Next, knocked out VNN1 via CRISPR/Cas9 system in the chicken Leghorn Male Hepatoma cell line. Detected the lipid deposition via oil red staining and analysis the content of triglycerides (TG), low-density lipoprotein-C (LDL-C), and highdensity lipoprotein-C (HDL-C) after VNN1 knockout in Leghorn Male Hepatoma cell line. Then we captured various differentially expressed genes (DEGs) between VNN1- modified LMH cells and original LMH cells by RNA-seq. Results: Firstly, fasting-induced expression of VNN1. Meanwhile, we successfully used the CRISPR/Cas9 system to achieve targeted mutations of the VNN1 in the chicken LMH cell line. Moreover, the expression level of VNN1 mRNA in LMH-KO-VNN1 cells decreased compared with that in the wild-type LMH cells (p<0.0001). Compared with control, lipid deposition was decreased after knockout VNN1 via oil red staining, meanwhile, the contents of TG and LDL-C were significantly reduced, and the content of HDL-C was increased in LMH-KO-VNN1 cells. Transcriptome sequencing showed that there were 1,335 DEGs between LMH-KO-VNN1 cells and original LMH cells. Of these DEGs, 431 were upregulated, and 904 were downregulated. Gene ontology analyses of all DEGs showed that the lipid metabolism-related pathways, such as fatty acid biosynthesis and long-chain fatty acid biosynthesis, were enriched. KEGG pathway analyses showed that “lipid metabolism pathway”, “energy metabolism”, and “carbohydrate metabolism” were enriched. A total of 76 DEGs were involved in these pathways, of which 29 genes were upregulated (such as cytochrome P450 family 7 subfamily A member 1, ELOVL fatty acid elongase 2, and apolipoprotein A4) and 47 genes were downregulated (such as phosphoenolpyruvate carboxykinase 1) by VNN1 knockout in the LMH cells. Conclusion: These results suggest that VNN1 plays an important role in coordinating lipid metabolism in the chicken liver.

ROBUST GAIN-SCHEDULING CONTROL OF DYNAMIC LATERAL OBSTACLE AVOIDANCE FOR CONNECTED AND AUTOMATED VEHICLES

Zhigen Nie,Zhongliang Li,Wanqiong Wang,Yufeng Lian,Rachid Outbib 한국자동차공학회 2023 International journal of automotive technology Vol.24 No.1

Dynamic trajectory planning (DTP) and Dynamic trajectory tracking (DTT) are the real-time mutual coupling in the process of the dynamic lateral obstacle avoidance (DLOA) of connected and automated vehicles (CAVs). Meanwhile, the varying velocity and acceleration of obstacle vehicles (OVs) increase the difficulties of DTP. Furthermore, the parameters perturbation in CAVs (such as mass and cornering stiffness), the varying velocities of CAVs and the signal disturbances, raise the difficulties of DTT. Therefore, the DLOA is challenging due to the interaction of the above multiple factors. To address the problem, this paper proposes a robust gain-scheduling control strategy of DLOA for CAVs. The strategy is divided into two modules namely DTP and DTT, and the two modules cooperate with each other in real time. In the module of DTP, the optimal trajectory considering the efficiency, passenger comfort and safety is real-time optimized in the dynamic safe limit which is real time predicted according to the information from CAVs and OVs. In the module of DTT, the real-time trajectory reference is tracked. Robust gain-scheduling control is realized to cope with variation of real-time trajectory reference, varying velocity, parameters perturbation and signal disturbances during the process of DLOA. The simulation results indicate that the strategy can effectively achieve DLOA maintaining the vehicle stability across various working conditions.

Generative Linguistic Steganography: A Comprehensive Review

Lingyun Xiang,Rong Wang,Zhongliang Yang,Yuling Liu 한국인터넷정보학회 2022 KSII Transactions on Internet and Information Syst Vol.16 No.3

Text steganography is one of the most imminent and promising research interests in the information security field. With the unprecedented success of the neural network and natural language processing (NLP), the last years have seen a surge of research on generative linguistic steganography (GLS). This paper provides a thorough and comprehensive review to summarize the existing key contributions, and creates a novel taxonomy for GLS according to NLP techniques and steganographic encoding algorithm, then summarizes the characteristics of generative linguistic steganographic methods properly to analyze the relationship and difference between each type of them. Meanwhile, this paper also comprehensively introduces and analyzes several evaluation metrics to evaluate the performance of GLS from diverse perspective. Finally, this paper concludes the future research work, which is more conducive to the follow-up research and innovation of researchers.

A Novel Low-cost Thin-film Flat Plate Photobioreactor for Microalgae Cultivation

Chenghu Yan,Qinghua Zhang,Shengzhang Xue,Zhongliang Sun,Xia Wu,Zhihui Wang,Yunming Lu,Wei Cong 한국생물공학회 2016 Biotechnology and Bioprocess Engineering Vol.21 No.1

In this study, a novel thin-film flat plate photobioreactor (FPPBR) mounted with baffles and a 61.2 m2 (2,000 L) photobioreactor system based on the FPPBR were developed. The flow of the fluid in the thinfilm photobioreactor was investigated by means of computational fluid dynamics (CFD). The cultivation of Chlorella sp. and Scenedesmus dimorphus in the thin-film FPPBR was carried out outdoors. The results showed that the flow of culture medium in different channels was uniform. In outdoor cultivation, the biomass productivity in the FPPBR with baffles was 25.2% higher than that in the FPPBR without baffles. In the pilot-scale FPPBR system, the maximum area productivity of Scenedesmus dimorphus reached 21.9 g/m2/day. When the service time of the photobioreactor was 1 and 3 years, the capital cost of the photobioreactor was 4.72 and 2.45 $ kgalgae, respectively. The results demonstrated that the thin-film FPPBR was cost effective, and it has the potential to be used for mass cultivation of microalgae.

Triterpenoid Saponins from Vaccaria segetalis

Shengmin Sang,Aina Lao,Hongcheng Wang,Zhongliang Chen,Jun Uzawa,Yasuo Fujimoto 한국생약학회 1998 Natural Product Sciences Vol.4 No.4

Two new triterpenoid saponins, named segetoside D and E, have been isolated from the seeds of Vaccaria segetalis. On the basis of chemical reactions and spectral data, structures of segetoside D and E have been established as: 28-O-[β-D-xylopyranosyl-(1→4)-α-L-rhamnopyranosyl-(1→2)]-[5-O-acetyl-α-arabinofuranosyl(1→3)]-[4-O-acetyl-β-D-fucopyranosyl]-quillaic acid-3-O-[β-D-galactopyranosyl(1→2)]-6-O-methyl ester-β-D-glucuronopyranoside and 28-O-[β-D-xylopyranosyl-(1→4)-α-L-rhamnopyranosyl-(1→2)]-[5-O-acetyl-α-arabinofuranosyl(1→3)]-[4-O-acetyl-β-D-fucopyranosyl]-quillaic acid -3-O-[β-D-galactopyranosyl(1→2)]-6-O-n-butyl ester-β-D-glucuronopyranoside, respectively.