Temporal expression profiling of long noncoding RNA and mRNA in the peripheral blood during porcine development

Gu, Yiren,Zhou, Rui,Jin, Long,Tao, Xuan,Zhong, Zhijun,Yang, Xuemei,Liang, Yan,Yang, Yuekui,Wang, Yan,Chen, Xiaohui,Gong, Jianjun,He, Zhiping,Li, Mingzhou,Lv, Xuebin Asian Australasian Association of Animal Productio 2020 Animal Bioscience Vol.33 No.5

Objective: We investigated the temporal expression profiles of long noncoding RNA (lncRNA) and mRNA in the peripheral blood of pigs during development and identified the lncRNAs that are related to the blood-based immune system. Methods: Peripheral blood samples were obtained from the pigs at 0, 7, 28, and 180 days and 2 years of age. RNA sequencing was performed to survey the lncRNA and mRNA transcriptomes in the samples. Short time-series expression miner (STEM) was used to show temporal expression patterns in the mRNAs and lncRNAs. Gene ontology and Kyoto encyclopedia of genes and genomes analyses were performed to assess the genes' biological relevance. To predict the functions of the identified lncRNAs, we extracted mRNAs that were nearby loci and highly correlated with the lncRNAs. Results: In total of 5,946 lncRNA and 12,354 mRNA transcripts were identified among the samples. STEM showed that most lncRNAs and mRNAs had similar temporal expression patterns during development, indicating the expressional correlation and functional relatedness between them. The five stages were divided into two classes: the suckling period and the late developmental stage. Most genes were expressed at low level during the suckling period, but at higher level during the late stages. Expression of several T-cell-related genes increased continuously during the suckling period, indicating that these genes are crucial for establishing the adaptive immune system in piglets at this stage. Notably, lncRNA TCONS-00086451 may promote blood-based immune system development by upregulating nuclear factor of activated T-cells cytoplasmic 2 expression. Conclusion: This study provides a catalog of porcine peripheral blood-related lncRNAs and mRNAs and reveals the characteristics and temporal expression profiles of these lncRNAs and mRNAs during peripheral blood development from the newborn to adult stages in pigs.

ZD1839 and Cisplatin Alone or in Combination for Treatment of a Nasopharyngeal Carcinoma Cell Line and Xenografts

Gu, Wei-Guang,Huang, Yan,Yuan, Zhong-Yu,Peng, Rou-Jun,Luo, Hai-Tao,He, Zhi-Ren,Wang, Shu-Sen Asian Pacific Journal of Cancer Prevention 2013 Asian Pacific journal of cancer prevention Vol.14 No.3

This study evaluated the effects of ZD1839, an orally active, selective epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor, on nasopharyngeal carcinoma (NPC) both in vitro and in vivo. Influence of ZD1839 alone or combined with cisplatin on the NPC cell line CNE2 was detected by MTT assay with flow cytometry assessment of cell cycle distribution and apoptosis rates. Nude mice NPC xenografts were also used to evaluate the effects of ZD1839 alone or combined with cisplatin. The Student's t test evaluated statistical significance. ZD1839 alone or combined with cisplatin inhibited CNE2 cell line proliferation. ZD1839 induced CNE2 cell cycle arrest in the G1 phase, and higher concentrations induced apoptosis. Xenograft tumors were significantly smaller when treated with 200 mg/kg ZD1839, cisplatin, or cisplatin combined with 100 mg/kg ZD1839 than untreated controls. ZD1839 (200 mg/kg) alone showed good tumor inhibition effects, reduction of tumor weights, and smaller tumor volume without loss of body weight. ZD1839 (200 mg/kg) might provide a good and effective therapeutic reagent for NPC.

Cadmium induces apoptosis in primary rat osteoblasts through caspase and mitogen-activated protein kinase pathways

Hong-Yan Zhao,Wei Liu,Yi Wang,Nannan Dai,Jian-Hong Gu,Yan Yuan,Xue-Zhong Liu,Jian-Chun Bian,Zong-Ping Liu 대한수의학회 2015 Journal of Veterinary Science Vol.16 No.3

Exposure to cadmium (Cd) induces apoptosis in osteoblasts (OBs); however, little information is available regarding the specific mechanismsof Cd-induced primary rat OB apoptosis. In this study, Cd reduced cell viability, damaged cell membranes and induced apoptosis in OBs. We observed decreased mitochondrial transmembrane potentials, ultrastructure collapse, enhanced caspase-3 activity, and increasedconcentrations of cleaved PARP, cleaved caspase-9 and cleaved caspase-3 following Cd treatment. Cd also increased the phosphorylationof p38-mitogen-activated protein kinase (MAPK), extracellular signal-regulated kinases (ERK)1/2 and c-jun N-terminal kinase (JNK) in OBs. Pretreatment with the caspase inhibitor, N-benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone, ERK1/2 inhibitor (U0126), p38 inhibitor(SB203580) and JNK inhibitor (SP600125) abrogated Cd-induced cell apoptosis. Furthermore, Cd-treated OBs exhibited signs of oxidativestress protection, including increased antioxidant enzymes superoxide dismutase and glutathione reductase levels and decreased formationof reactive oxygen species. Taken together, the results of our study clarified that Cd has direct cytotoxic effects on OBs, which are mediatedby caspase- and MAPK pathways in Cd-induced apoptosis of OBs.

Regulation of matrix metalloproteinase-9 protein expression by 1α , 25-(OH)2D3 during osteoclast differentiation

Jian-Hong Gu,Xi-Shuai Tong,Guohong Chen,Xue-Zhong Liu,Jian-Chun Bian,Yan Yuan,Zong-Ping Liu 대한수의학회 2014 Journal of Veterinary Science Vol.15 No.1

To investigate 1α,25-(OH)2D3 regulation of matrixmetalloproteinase-9 (MMP-9) protein expression duringosteoclast formation and differentiation, receptor activator ofnuclear factor κB ligand (RANKL) and macrophagecolony-stimulating factor (M-CSF) were administered toinduce the differentiation of RAW264.7 cells into osteoclasts. The cells were incubated with different concentrations of1α,25-(OH)2D3 during culturing, and cell proliferation wasmeasured using the methylthiazol tetrazolium method. Osteoclast formation was confirmed using tartrate-resistantacid phosphatase (TRAP) staining and assessing bone lacunarresorption. MMP-9 protein expression levels were measuredwith Western blotting. We showed that 1α,25-(OH)2D3inhibited RAW264.7 cell proliferation induced by RANKLand M-CSF, increased the numbers of TRAP-positiveosteoclasts and their nuclei, enhanced osteoclast boneresorption, and promoted MMP-9 protein expression in aconcentration-dependent manner. These findings indicatethat 1α,25-(OH)2D3 administered at a physiological relevantconcentration promoted osteoclast formation and couldregulate osteoclast bone metabolism by increasing MMP-9protein expression during osteoclast differentiation.

Decreased Expression of FADS1 Predicts a Poor Prognosis in Patients with Esophageal Squamous Cell Carcinoma

Du, Yong,Yan, Shu-Mei,Gu, Wan-Yi,He, Fan,Huang, Li-Yun,Li, Mei,Yuan, Yan,Chen, Ren-Hui,Zhong, Qian,Li, Man-Zhi,Li, Yong,Zeng, Mu-Sheng Asian Pacific Journal of Cancer Prevention 2015 Asian Pacific journal of cancer prevention Vol.16 No.12

FADS1 (fatty acid desaturase 1) plays a crucial role in fatty acid metabolism, and it was recently reported to be involved in tumorigenesis. However, the role of FADS1 expression in esophageal squamous cell carcinoma (ESCC) remains unknown. In the current study, we investigated the expression and clinical pathologic and prognostic significance of FADS1 in ESCC. Immunohistochemical analyses revealed that 58.2% (146/251) of the ESCC tissues had low levels of FADS1 expression, whereas 41.8% (105/251) exhibited high levels of FADS1 expression. In positive cases, FADS1 expression was detected in the cytoplasm of cells. Correlation analyses demonstrated that FADS1 expression was significantly correlated with tumor location (p=0.025) but not with age, gender, histological grade, tumor status, nodal status or TNM staging. Furthermore, patients with tumors expressing high levels of FADS1had a longer disease-free survival time (p<0.001) and overall survival time (p <0.001). Univariate and multivariate analyses revealed that, along with nodal status, FADS1 expression was an independent and significant predictive factor (p<0.001). In conclusion, our study suggested that FADS1 might be a valuable biomarker and potential therapeutic target for ESCC.

Tensile and Creep Deformation of a Newly Developed Ni-Fe-Based Superalloy for 700 °C Advanced Ultra-Supercritical Boiler Applications

Y. Yuan,Z. H. Zhong,Z.S. Yu,H. F. Yin,Y. Y. Dang,X. B. Zhao,Z. Yang,J. T. Lu,J. B. Yan,Y. Gu 대한금속·재료학회 2015 METALS AND MATERIALS International Vol.21 No.4

A new Ni-Fe-based superalloy, HT-X, has been developed for applications in 700 °C advanced ultra-supercritical (A-USC) boilers. The HT-X alloy is subjected to various heat treatments. Tensile tests are conducted at room temperature (RT), 700 °C and 750 °C. Creep tests are carried out under conditions of 700 °C/300 MPa and 750 °C/150 MPa. After aging treatment, the yield strength of the HT-X alloy at RT and 750 °C is 787 MPa and 624 MPa, respectively. When additional thermal exposure at 750 °C for 5400 h is applied, the yield strength is decreased to 656 MPa at RT and 480 MPa at 700 °C. For an aged specimen, the a/2<110> dislocation shearing process occurs when tensile testing is conducted at RT and 750 °C. As the γ' precipitate size increases in the specimen that is thermally exposed at 750 °C for 5400 h, Orowan bowing is the dominant dislocation process, and stacking faults develop in the γ' precipitates at both RT and 700 °C. Dislocation slip combined with climb is the dominant mechanism under the creep testing conditions. The factors that affect the mechanical properties and deformation mechanisms are discussed.

Inhibitory effects of osteoprotegerin on osteoclast formation and function under serum-free conditions

Ying-Xiao Fu,Jian-Hong Gu,Yi-Ran Zhang,Xi-Shuai Tong,Hong-Yan Zhao,Yan Yuan,Xue-Zhong Liu,Jian-Chun Bian,Zong-Ping Liu 대한수의학회 2013 Journal of Veterinary Science Vol.14 No.4

The purpose of this study was to determine whether osteoprotegerin (OPG) could affect osteoclat differentiation and activation under serum-free conditions. Both duck embryo bone marrow cells and RAW264.7 cells were incubated with macrophage colony stimulatory factor (M-CSF) and receptor activator for nuclear factor κB ligand (RANKL) in serum-free medium to promote osteoclastogenesis. During cultivation, 0,10, 20, 50, and 100 ng/mL OPG were added to various groups of cells. Osteoclast differentiation and activation were monitored via tartrate-resistant acid phosphatase (TRAP) staining,filamentous-actin rings analysis, and a bone resorption assay. Furthermore, the expression osteoclast-related genes, such as TRAP and receptor activator for nuclear factor κB (RANK),that was influenced by OPG in RAW264.7 cells was examined using real-time polymerase chain reaction. In summary,findings from the present study suggested that M-CSF with RANKL can promote osteoclast differentiation and activation,and enhance the expression of TRAP and RANK mRNA in osteoclasts. In contrast, OPG inhibited these activities under serum-free conditions.

Involvement of the Ca2+ signaling pathway in osteoprotegerin inhibition of osteoclast differentiation and maturation

Ying-Xiao Fu,Jian-Hong Gu,Yi Wang,Yan Yuan,Xue-Zhong Liu,Jian-Chun Bian,Zong-Ping Liu 대한수의학회 2015 Journal of Veterinary Science Vol.16 No.2

The purpose of this study was to determine whether the Ca2+ signaling pathway is involved in the ability of osteoprotegerin (OPG) to inhibit osteoclast differentiation and maturation. RAW264.7 cells were incubated with macrophage colony-stimulating factor (M-CSF) + receptor activator of nuclear factor-κB ligand (RANKL) to stimulate osteoclastogenesis and then treated with different concentrations of OPG, an inhibitor of osteoclast differentiation. The intracellular Ca2+ concentration [Ca2+]i and phosphorylation of Ca2+/calmodulin-dependent protein kinase II (CaMKII) in the different treatment groups were measured by flow cytometry and Western blotting, respectively. The results confirmed that M-CSF + RANKL significantly increased [Ca2+]i and CaMKII phosphorylation in osteoclasts (p < 0.01), and that these effects were subsequently decreased by OPG treatment. Exposure to specific inhibitors of the Ca2+ signaling pathway revealed that these changes varied between the different OPG treatment groups. Findings from the present study indicated that the Ca2+ signaling pathway is involved in both the regulation of osteoclastogenesis as well as inhibition of osteoclast differentiation and activation by OPG.

Lethal and sublethal effects of sulfoxaflor on the small brown planthopper Laodelphax striatellus

Lu Xu,Chun-Qing Zhao,Ya-Nan Zhang,Ying Liud,,Zhong-Yan Gu 한국응용곤충학회 2016 Journal of Asia-Pacific Entomology Vol.19 No.3

Laodelphax striatellus (Fallén), as an important pest of gramineous crops, has developed resistance to multiple classes of insecticides, impairing control efficiency. However, the application of insecticides is still the main controlmeasure for it.Herein, the lethal effects of seven insecticides aswell as sulfoxaflor,which has been introduced and registered for controlling L. striatellus in China, were investigated. The acute toxicity of these insecticides on the L. striatellus adultswas rankedas sulfoxaflor N abamectin N dinotefuran N emamectin benzoate N ethofenprox N imidacloprid N chlorantraniliprole N chlorpyrifos. The toxicity of sulfoxaflor against the adultswas highestwith LD50 at 1.07–1.09 ng/insect. In addition, the sublethal effects of lower lethal dose LD3 (0.06 ng/insect), low lethal dose LD10 (0.15 ng/insect) and moderate lethal dose LD30 (0.49 ng/insect) of sulfoxaflor for L. striatellus were assessed as well. Both LD10 and LD30 induced slower nymphal development period, shorter oviposition period and longer pre-oviposition period in L. striatellus. The LD30 also shortened the longevity of females. Hormesis on fecunditywas observed in L. striatellus exposed to LD3. Therefore, the net reproductive rate (R0) was increased by LD3. The intrinsic rate of increase (rm) was reduced by LD10 and LD30 while mean generation time(T) and doubling time (DT) were prolonged. The acute sulfoxaflor doses in its toxicological properties need to be considered when develop L. striatellus control strategy with sulfoxaflor. These results demonstrate that sulfoxaflor is a valid candidate for L. striatellus management.

Multiple down-regulated cytochrome P450 monooxygenase genes contributed to synergistic interaction between chlorpyrifos and imidacloprid against Nilaparvata lugens

Lu Xue,Guanghua Luo,Yang Sun,Shuijin Huang,De-Jin Xu,Guang-Chun Xu,Zhao-Jun Han,Zhong-Yan Gu,Ya-Nan Zhang 한국응용곤충학회 2020 Journal of Asia-Pacific Entomology Vol.23 No.1

Insecticide mixtures are an effective strategy in pest resistance management. The synergistic chlorpyrifos and imidacloprid mixture could significantly increase toxicity against rice pest, Nilaparvata lugens, despite their high levels of resistance. However, synergism mechanisms to explain this phenomenon remain unknown. Chlorpyrifos and imidacloprid at a 1:0.5 ratio showed significant synergism on N. lugens with a combination index value of 0.18 after topical exposure. We constructed a genetic database of the genes expressed in individual and synergistic chlorpyrifos and imidacloprid treatments of N. lugens using Illumina Hiseq™ X Ten, and 17 co-downregulated genes putatively involved in synergism were detected by comparative transcriptome analyses. Expression patterns of the 17 candidate synergistic genes matched with transcriptome sequencing data by quantitative real-time PCR analyses. Feeding of dsRNAs further reduced the expression levels of 10 of these candidate synergistic genes (from 1.68 to 4.13-fold). Nymphs fed with only dsRNAs of CYP4DE1, CYP6AY1v2, CYP353D1, and CYP439A1 experienced more high mortality rates (81.45–90.34%) to improve synergism between chlorpyrifos and imidacloprid. Multiple reductive expressed P450 genes were potentially associated with synergism of a mixture of chlorpyrifos and imidacloprid, as confirmed by comparative transcriptome analyses and RNAi assays. Our findings suggested that synergistic interactions between chlorpyrifos and imidacloprid might be controlled by P450s.