Relationship between porcine miR-20a and its putative target low-density lipoprotein receptor based on dual luciferase reporter gene assays

Yueyun Ding,Shujiao Zhu,Chaodong Wu,Li Qian,DengTao Li,Li Wang,Yuanlang Wang,Wei Zhang,Min Yang,Jian Ding,Xudong Wu,Xiao-Dong Zhang,Yafei Gao,Zongjun Yin 아세아·태평양축산학회 2019 Animal Bioscience Vol.32 No.7

Objective: Mutations in low-density lipoprotein receptor (LDLR), which encodes a critical protein for cholesterol homeostasis and lipid metabolism in mammals, are involved in cardiometabolic diseases, such as familial hypercholesterolemia in pigs. Whereas microRNAs (miRNAs) can control LDLR regulation, their involvement in circulating cholesterol and lipid levels with respect to cardiometabolic diseases in pigs is unclear. We aimed to identify and analyze LDLR as a potential target gene of SSC-miR-20a. Methods: Bioinformatic analysis predicted that porcine LDLR is a target of SSC-miR-20a. Wild-type and mutant LDLR 3′-untranslated region (UTR) fragments were generated by polymerase chain reaction (PCR) and cloned into the pGL3-Control vector to construct pGL3 Control LDLR wild-3′-UTR and pGL3 Control LDLR mutant-3′-UTR recombinant plasmids, respectively. An miR-20a expression plasmid was constructed by inserting the porcine pre-miR-20a-coding sequence between the HindIII and BamHI sites in pMR-mCherry, and constructs were confirmed by sequencing. HEK293T cells were co-transfected with the miR-20a expression or pMR-mCherry control plasmids and constructs harboring the corresponding 3′-UTR, and relative luciferase activity was determined. The relative expression levels of miR-20a and LDLR mRNA and their correlation in terms of expression levels in porcine liver tissue were analyzed using reverse-transcription quantitative PCR. Results: Gel electrophoresis and sequencing showed that target gene fragments were successfully cloned, and the three recombinant vectors were successfully constructed. Compared to pMR-mCherry, the miR-20a expression vector significantly inhibited wild-type LDLR-3′-UTR-driven (p<0.01), but not mutant LDLR-3′-UTR-driven (p>0.05), luciferase reporter activity. Further, miR-20a and LDLR were expressed at relatively high levels in porcine liver tissues. Pearson correlation analysis revealed that porcine liver miR-20a and LDLR levels were significantly negatively correlated (r = –0.656, p<0.05). Conclusion: LDLR is a potential target of miR-20a, which might directly bind the LDLR 3′-UTR to post-transcriptionally inhibit expression. These results have implications in understanding the pathogenesis and progression of porcine cardiovascular diseases.

Relationship among porcine lncRNA TCONS_00010987, miR-323, and leptin receptor based on dual luciferase reporter gene assays and expression patterns

Ding, Yueyun,Qian, Li,Wang, Li,Wu, Chaodong,Li, DengTao,Zhang, Xiaodong,Yin, Zongjun,Wang, Yuanlang,Zhang, Wei,Wu, Xudong,Ding, Jian,Yang, Min,Zhang, Liang,Shang, Jinnan,Wang, Chonglong,Gao, Yafei Asian Australasian Association of Animal Productio 2020 Animal Bioscience Vol.33 No.2

Objective: Considering the physiological and clinical importance of leptin receptor (LEPR) in regulating obesity and the fact that porcine LEPR expression is not known to be controlled by lncRNAs and miRNAs, we aim to characterize this gene as a potential target of SSC-miR-323 and the lncRNA TCONS_00010987. Methods: Bioinformatics analyses revealed that lncRNA TCONS_00010987 and LEPR have SSC-miR-323-binding sites and that LEPR might be a target of lncRNA TCONS_00010987 based on cis prediction. Wild-type and mutant TCONS_00010987-target sequence fragments and wild-type and mutant LEPR 3'-UTR fragments were generated and cloned into pmiRRB-REPORT^TM-Control vectors to construct respective recombinant plasmids. HEK293T cells were co-transfected with the SSC-miR-323 mimics or a negative control with constructs harboring the corresponding binding sites and relative luciferase activities were determined. Tissue expression patterns of lncRNA TCONS_00010987, SSC-miR-323, and LEPR in Anqing six-end-white (AQ, the obese breed) and Large White (LW, the lean breed) pigs were detected by real-time quantitative polymerase chain reaction; backfat expression of LEPR protein was detected by western blotting. Results: Target gene fragments were successfully cloned, and the four recombinant vectors were constructed. Compared to the negative control, SSC-miR-323 mimics significantly inhibited luciferase activity from the wild-type TCONS_00010987-target sequence and wild-type LEPR-3'-UTR (p<0.01 for both) but not from the mutant TCONS_00010987-target sequence and mutant LEPR-3'-UTR (p>0.05 for both). Backfat expression levels of TCONS_00010987 and LEPR in AQ pigs were significantly higher than those in LW pigs (p<0.01), whereas levels of SSC-miR-323 in AQ pigs were significantly lower than those in LW pigs (p<0.05). LEPR protein levels in the backfat tissues of AQ pigs were markedly higher than those in LW pigs (p<0.01). Conclusion: LEPR is a potential target of SSC-miR-323, and TCONS_00010987 might act as a sponge for SSC-miR-323 to regulate LEPR expression.

A Lifetime-Preserving and Delay-Constrained Data Gathering Tree for Unreliable Sensor Networks

( Yanjun Li ),( Yueyun Shen ),( Kaikai Chi ) 한국인터넷정보학회 2012 KSII Transactions on Internet and Information Syst Vol.6 No.12

A tree routing structure is often adopted for many-to-one data gathering and aggregation in sensor networks. For real-time scenarios, considering lossy wireless links, it is an important issue how to construct a maximum-lifetime data gathering tree with delay constraint. In this work, we study the problem of lifetime-preserving and delay-constrained tree construction in unreliable sensor networks. We prove that the problem is NP-complete. A greedy approximation algorithm is proposed. We use expected transmissions count (ETX) as the link quality indicator, as well as a measure of delay. Our algorithm starts from an arbitrary least ETX tree, and iteratively adjusts the hierarchy of the tree to reduce the load on bottleneck nodes by pruning and grafting its sub-tree. The complexity of the proposed algorithm is O(N⁴). Finally, extensive simulations are carried out to verify our approach. Simulation results show that our algorithm provides longer lifetime in various situations compared to existing data gathering schemes.

Evaluation and modeling of methyl green adsorption from aqueous solutions using loofah fibers

Xiaonan Tang,Yueyun Li,Runhai Chen,Fanlian Min,Juanjuan Yang,Yunhui Dong 한국화학공학회 2015 Korean Journal of Chemical Engineering Vol.32 No.1

Loofah fiber, an economical adsorbent material, was first developed for the methyl green (MG) removalfrom aqueous solutions. The loofah fibers were characterized by SEM, FTIR, N2-BET and the potentiometric titration. The pH, contact time and temperature were examined extensively. The adsorption of MG on loofah fiber increasedvery quickly in the pH range 3.0 to 7.0, remaining a high level at pH>7.0. The kinetics of adsorption of MG on theloofah fiber was proved to coincide with pseudo-second-order kinetic models (r2>0.99) very well. Langmuir isothermwas demonstrated to fit the experimental data better than Freundlich isotherm model. Monolayer adsorption capacityincreased with the increase of temperature. Thermodynamic constants were evaluated, and the results indicated thatMG adsorption onto loofah fiber was feasible, spontaneous and endothermic. The high removal efficiency of MG onloofah fiber suggested that the loofah fiber was suitable material in MG pollution cleanup.

Cyanide- and Phenoxo-Bridged Heterobimetallic Fe(III)-Mn(III) Coordination Polymer: Synthesis, Crystal Structures and Magnetic Properties

Zhang, Daopeng,Kong, Lingqian,Li, Yueyun,Wang, Ping,Chen, Xia Korean Chemical Society 2014 Bulletin of the Korean Chemical Society Vol.35 No.9

Two two-dimensional cyanide- and phenoxo-bridged heterometallic M(II)-Mn(III) (M = Ni, Pd) coordination polymers $\{[Mn(saltmen)]_4[Ni(CN)_4]\}(ClO_4)_2{\cdot}CH_3OH{\cdot}H_2O$ (1) and $\{[Mn(saltmen)]_4[Pd(CN)_4]\}(ClO_4)_2{\cdot}CH_3CN{\cdot}H_2O$ (2) ($saltmen^{2-}$ = N,N'-(1,1,2,2-tetramethylethylene)bis(salicylideneaminato)dianion) have been obtained by using $K_2[M(CN)_4]$ as building blocks and a salen-tpye Schiff-base manganese(III) compound as assembling segment. Single X-ray analysis reveals their isostrutural cyanide-bridged $MMn_4$ pentanuclear cationic structure. The four Schiff base manganese units of the pentanuclear entity are self-complementary through the phenoxo oxygen atoms from the neighboring complex, therefore forming cyanide- and phenoxo-bridged 2D sheet-like structure. Investigation over magnetic susceptibilities reveals the overall ferromagnetic coupling between the adjacent Mn(III) ions bridged by the phenoxo oxygen atoms with J = 2.13 and $2.21cm^{-1}$ for complexes 1 and 2, respectively.

Cyanide- and Phenoxo-Bridged Heterobimetallic Fe(III)-Mn(III) Coordination Polymer: Synthesis, Crystal Structures and Magnetic Properties

Daopeng Zhang,Lingqian Kong,Yueyun Li,Ping Wang,Xia Chen 대한화학회 2014 Bulletin of the Korean Chemical Society Vol.35 No.9

Two two-dimensional cyanide- and phenoxo-bridged heterometallic M(II)-Mn(III) (M = Ni, Pd) coordination polymers {[Mn(saltmen)]4[Ni(CN)4]}(ClO4)2·CH3OH·H2O (1) and {[Mn(saltmen)]4[Pd(CN)4]}(ClO4)2· CH3CN·H2O (2) (saltmen2– = N,N'-(1,1,2,2-tetramethylethylene)bis(salicylideneaminato)dianion) have been obtained by using K2[M(CN)4] as building blocks and a salen-tpye Schiff-base manganese(III) compound as assembling segment. Single X-ray analysis reveals their isostrutural cyanide-bridged MMn4 pentanuclear cationic structure. The four Schiff base manganese units of the pentanuclear entity are self-complementary through the phenoxo oxygen atoms from the neighboring complex, therefore forming cyanide- and phenoxobridged 2D sheet-like structure. Investigation over magnetic susceptibilities reveals the overall ferromagnetic coupling between the adjacent Mn(III) ions bridged by the phenoxo oxygen atoms with J = 2.13 and 2.21 cm−1 for complexes 1 and 2, respectively.

Systematic Approach for Design of Broadband, High Efficiency, High Power RF Amplifiers

Seyed Alireza Mohadeskasaei,Jianwei An,Yueyun Chen,Zhi Li,Sani Umar Abdullahi,Tie Sun 한국전자통신연구원 2017 ETRI Journal Vol.39 No.1

This paper demonstrates a systematic approach for the design of broadband, high efficiency, high power, Class-AB RF amplifiers with high gain flatness. It is usually difficult to simultaneously achieve a high gain flatness and high efficiency in a broadband RF power amplifier, especially in a high power design. As a result, the use of a computer-aided simulation is most often the best way to achieve these goals; however, an appropriate initial value and a systematic approach are necessary for the simulation results to rapidly converge. These objectives can be accomplished with a minimum of trial and error through the following techniques. First, signal gain variations are reduced over a wide bandwidth using a proper pre-matching network. Then, the source and load impedances are satisfactorily obtained from small-signal and load-pull simulations, respectively. Finally, two high-order Chebyshev low-pass filters are employed to provide optimum input and output impedance matching networks over a bandwidth of 100 MHz–500 MHz. By using an EM simulation for the substrate, the simulation results were observed to be in close agreement with the measured results.

Removal of Co(II) from aqueous solutions by sulfonated magnetic multi-walled carbon nanotubes

Juanjuan Yang,Yunhui Dong,Jun Li,Zhengjie Liu,Fanlian Min,Yueyun Li 한국화학공학회 2015 Korean Journal of Chemical Engineering Vol.32 No.11

Sulfonated magnetic multi-walled carbon nanotubes (SMMWCNTs) were applied in the sorption of Co(II) from aqueous solutions. The SMMWCNTs were prepared and characterized by scanning electron microscope (SEM), Fourier transform infrared (FTIR), and X-ray diffractometer (XRD) test. A large number of influencing factors to the sorption process were investigated, such as pH, ionic strength, contact time, cations, anions, humic acid (HA), fulvic acid (FA) and temperature. The results indicated that the Co(II) sorption was strongly controlled by the pH and ionic strength. Moreover, foreign anions, such as F−, Cl− and Br−, had an obvious effect on the sorption process, which depended on the electronegativity of the anions. On the other hand, cations restrained sorption strongly, such as Mg2+ and Ca2+. The existence of HA/FA enhanced sorption process at pH<8 while weakened at pH>8. As revealed by the sorption results, the Langmuir adsorption model was more favorable than the Freundlich adsorption model, and the pseudo-second-order model could fit the data much better than the pseudo-first-order. The thermodynamic analysis suggested that sorption was spontaneous and endothermic. What’s more, the stability experiments of the SMMWCNTs showed that SMMWCNTs could maintain excellent magnetic stability and dispersion stability. Thus, this SMMWCNTs sorbent was believed to be a promising material for the selective removal of Co(II) from heavy metal-containing wastewater.