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.

Adverse effect of DEHP exposure on the serum insulin level of Balb/c mice

Jing Wang,Jinquan Li,Kashif Rafiq Zahid,Kun Wang,Yan Qian,Ping Ma,Shumao Ding,Xu Yang,Xianliang Wang,Xianliang Wang,Xu Yang 대한독성 유전단백체 학회 2016 Molecular & cellular toxicology Vol.12 No.1

Di(2-ethylhexyl) phthalate (DEHP) is a common indoor pollutant in the world, which may cause lots of harmful effects in human including diabetes according to epidemiological studies. To explore the underlying role of DEHP in diabetes-like symptoms, Balb/c mice were chose to be the experimental animals in this paper. They were separated as eight groups as follows: (1) saline+normal diet (vehicle control), (2) 10 mg/kg.day DEHP+normal diet, (3) 50 mg/kg.day DEHP+normal diet, (4) 250 mg/kg.day DEHP+normal diet, (5) streptozotocin (STZ)+high fat diet (diabetes model), (6) 10 mg/kg.day DEHP+ STZ+high fat diet, (7) 50 mg/kg.day DEHP+STZ +high fat diet, (8) 250 mg/kg.day DEHP+STZ+high fat diet. The biomarkers of this experiment include four groups: (1) general indicates: body weight and drinking water, (2) blood biomarkers: serum insulin and fasting glucose, (3) pathological examination: pancreas section and kidney section, and (4) biomarkers of oxidative stress: reactive oxygen species (ROS) and malondialdehyde (MDA) in liver cells. Our study results demonstrate that: (1) at our treatment levels DEHP cannot directly induce diabetes, but reduce serum insulin level in DEHP-exposed non-STZ-treated animals, (2) pathological examination finds that these is a dose-dependent damage in the pancreas in DEHP- exposed STZ-treated groups, and (3) the oxidative mechanism may be involved in this pathological process.

Chicken serum uric acid level is regulated by glucose transporter 9

Ding Xuedong,Peng Chenglu,Li Siting,Li Manman,Li Xinlu,Wang Zhi,Li Yu,Wang Xichun,Li Jinchun,wu Jinjie 아세아·태평양축산학회 2021 Animal Bioscience Vol.34 No.4

Objective: Glucose transporter 9 (GLUT9) is a uric acid transporter that is associated with uric absorption in mice and humans; but it is unknown whether GLUT9 involves in chicken uric acid regulation. This experiment aimed to investigate the chicken GLUT9 expression and serum uric acid (SUA) level. Methods: Sixty chickens were divided into 4 groups (n = 15): a control group (NC); a sulfonamide-treated group (SD) supplemented with sulfamonomethoxine sodium via drinking water (8 mg/L); a fishmeal group (FM) supplemented with 16% fishmeal in diet; and a uric acid-injection group (IU), where uric acid (250 mg/kg) was intraperitoneally injected once a day. The serum was collected weekly to detect the SUA level. Liver, kidney, jejunum, and ileum tissues were collected to detect the GLUT9 mRNA and protein expression. Results: The results showed in the SD and IU groups, the SUA level increased and GLUT9 expression increased in the liver, but decreased in the kidney, jejunum, and ileum. In the FM group, the SUA level decreased slightly and GLUT9 expression increased in the kidney, but decreased in the liver, jejunum, and ileum. Correlation analysis revealed that liver GLUT9 expression correlated positively, and renal GLUT9 expression correlated negatively with the SUA level. Conclusion: These results demonstrate that there may be a feedback regulation of GLUT9 in the chicken liver and kidney to maintain the SUA balance; however, the underlying mechanism needs to be investigated in future studies. Objective: Glucose transporter 9 (GLUT9) is a uric acid transporter that is associated with uric absorption in mice and humans; but it is unknown whether GLUT9 involves in chicken uric acid regulation. This experiment aimed to investigate the chicken GLUT9 expression and serum uric acid (SUA) level.Methods: Sixty chickens were divided into 4 groups (n = 15): a control group (NC); a sulfonamide-treated group (SD) supplemented with sulfamonomethoxine sodium via drinking water (8 mg/L); a fishmeal group (FM) supplemented with 16% fishmeal in diet; and a uric acid-injection group (IU), where uric acid (250 mg/kg) was intraperitoneally injected once a day. The serum was collected weekly to detect the SUA level. Liver, kidney, jejunum, and ileum tissues were collected to detect the GLUT9 mRNA and protein expression.Results: The results showed in the SD and IU groups, the SUA level increased and GLUT9 expression increased in the liver, but decreased in the kidney, jejunum, and ileum. In the FM group, the SUA level decreased slightly and GLUT9 expression increased in the kidney, but decreased in the liver, jejunum, and ileum. Correlation analysis revealed that liver GLUT9 expression correlated positively, and renal GLUT9 expression correlated negatively with the SUA level.Conclusion: These results demonstrate that there may be a feedback regulation of GLUT9 in the chicken liver and kidney to maintain the SUA balance; however, the underlying mechanism needs to be investigated in future studies.

Analysis on stability of an Arch Dam with Interlayer Shear Zones

Ding Zelin,Wang Qianqian,Wang Jing 대한토목학회 2016 KSCE JOURNAL OF CIVIL ENGINEERING Vol.20 No.6

This paper focuses on the stability of arch dam abutment with interlayer shear zones, presents the experimental analysis on Lizhou Arch Dam, and implements comprehensive method by combining geo-mechanical model test with the finite element method to analyze the impact of interlayer shear zones on the stability of dam abutment. Firstly, 3D geo-mechanical model test is conducted and the model shows the impact of interlayer shear zones on stability of arch dam. According to the deformation behaviors and eventual failure form of rock mass and structure planes in dam abutment and dam foundation, the impact of interlayer shear zones unstable failure on the safety coefficient of arch dam is analyzed. Secondly, the FEM (Finite Element Method) is used for supplementary verification. As a conclusion of the analysis: Interlayer shear zones fj1 - fj4 are developing towards riverbed in the rock mass of the dam abutment of Lizhou Arch Dam, cutting through the rock mass of dam abutment and forming a bottom slide face. In the dam abutment, fault f5 and fracture Lp285 have developed and formed a side fracture plane by intersecting the interlayer shear zone. Under the effect of the resultant force of the arch abutment, with the interlayer shear zone of inferior properties as the bottom slide face, the rock mass of dam abutment slides out in the form of steps or drawers, and the stability of the dam and the foundation is influenced.

Recent advances of Na3V2(PO4)3 as cathode for rechargeable zinc-based batteries

Ding Kunpeng,Jiang Tian,Peng Jian,Wang Peng,Gou Wenshan,Xu Qingyu,Fan Qi,Wang Wei,Sun Yueming 한국탄소학회 2023 Carbon Letters Vol.33 No.4

Rechargeable zinc-based batteries (RZBs) with the advantages of high safety, low cost, abundant resources and environmental friendliness, are considered as advanced secondary battery systems that can be applied to large-scale energy storage. As an important cathode material for RZBs, NASICON-type Na3V2(PO4)3 (NVP) possesses three-dimensional and large-scale ion channels that facilitate the rapid diffusion of Zn2+, and has a higher average operating voltage compared with other vanadium-based compounds, thus exhibiting the possibility of realizing RZBs with high energy density. However, NVP still has some problems, such as poor electronic conductivity and spontaneous dissolution in aqueous solution. The sluggish kinetics of Zn2+ (de)intercalation in NVP and dendritic growth on the Zn anode also contribute to the poor rate performance and short cycle life of the batteries. In this review, optimization strategies for the electrochemical performance of RZBs with NVP as cathode are systematically elaborated, including modification of NVP cathode and optimization of electrolyte. Several mainstream energy storage mechanisms and analysis methods in this battery system are sorted out and summarized. On this basis, the development direction of NVP–RZB system is further prospected.

Adaptive guided salp swarm algorithm with velocity clamping mechanism for solving optimization problems

Wang Zongshan,Ding Hongwei,Wang Jie,Hou Peng,Li Aishan,Yang Zhijun,Hu Xiang 한국CDE학회 2022 Journal of computational design and engineering Vol.9 No.6

Salp swarm algorithm (SSA) is a well-established population-based optimizer that exhibits strong exploration ability, but slow convergence and poor exploitation capability. In this paper, an endeavour is made to enhance the performance of the basic SSA. The new upgraded version of SSA named as ‘adaptive strategy-based SSA (ABSSA) algorithm’ is proposed in this paper. First, the exploratory scope and food source navigating commands of SSA are enriched using the inertia weight and boosted global best-guided mechanism. Next, a novel velocity clamping strategy is designed to efficiently stabilize the balance between the exploration and exploitation operations. In addition, an adaptive conversion parameter tactic is designed to modify the position update equation to effectively intensify the local exploitation competency and solution accuracy. The effectiveness of the proposed ABSSA algorithm is verified by a series of problems, including 23 classical benchmark functions, 29 complex optimization problems from CEC 2017, and five engineering design tasks. The experimental results show that the developed ABSSA approach performs significantly better than the standard SSA and other competitors. Moreover, ABSSA is implemented to handle path planning and obstacle avoidance (PPOA) tasks in autonomous mobile robots and compared with some swarm intelligent approach-based path planners. The experimental results indicate that the ABSSA-based PPOA method is a reliable path planning algorithm.

Excessive mechanical strain accelerates intervertebral disc degeneration by disrupting intrinsic circadian rhythm

Ding Sheng-Long,Zhang Tai-Wei,Zhang Qi-Chen,Ding Wang,Li Ze-Fang,Han Guan-Jie,Bai Jin-Song,Li Xi-Lei,Dong Jian,Wang Hui-Ren,Jiang Li-Bo 생화학분자생물학회 2021 Experimental and molecular medicine Vol.53 No.-

Night shift workers with disordered rhythmic mechanical loading are more prone to intervertebral disc degeneration (IDD). Our results showed that circadian rhythm (CR) was dampened in degenerated and aged NP cells. Long-term environmental CR disruption promoted IDD in rats. Excessive mechanical strain disrupted the CR and inhibited the expression of core clock proteins. The inhibitory effect of mechanical loading on the expression of extracellular matrix genes could be reversed by BMAL1 overexpression in NP cells. The Rho/ROCK pathway was demonstrated to mediate the effect of mechanical stimulation on CR. Prolonged mechanical loading for 12 months affected intrinsic CR genes and induced IDD in a model of upright posture in a normal environment. Unexpectedly, mechanical loading further accelerated the IDD in an Light-Dark (LD) cycle-disrupted environment. These results indicated that intrinsic CR disruption might be a mechanism involved in overloading-induced IDD and a potential drug target for night shift workers.

Construction and analysis of Sip1Aa insecticidal protein random recombination library

Ding Yue-ming,Wang Jing,Wang Lin,Gao Ji-Guo,Liu Rong-Mei,Li Hai-Tao 한국응용곤충학회 2021 Journal of Asia-Pacific Entomology Vol.24 No.2

The Sip1Aa protein from Bacillus thuringiensis is highly toxic to Colaphellus bowringi Baly. In order to obtain mutant proteins with higher insecticidal activity, a random recombinant library of Sip1Aa protein was con structed using error-prone PCR. A total number of 100 positive transformants were randomly selected for sequence determination, and 25 mutants (M1 to M25) were selected and expressed the respective Sip1Aa mu tants. These Sip1Aa variants had a total of 29 base mutations, with an average of 1.2 base mutations per mutant. Compared with that of the wild-type Sip1Aa protein, the insecticidal activity of the mutants M1 (A31G, Y118C, D227E), M5 (K168R) and M21 (I307T) was significantly decreased, with and LC 50 values 4 to 6 times higher than the Sip1Aa protein. The mutant M8 (R174S) showed increase in the insecticidal activity against the Colaphellus bowringi Baly was obtained, with an LC 50 value 4-fold less than the Sip1Aa protein. The results of this study provide reference for the molecular modification of Sip1Aa protein and the study of key sites of its insecticidal activity.

Genetic Diversity of a Chinese Native Chicken Breed, Bian Chicken, Based on Twenty-nine Microsatellite Markers

Ding, Fu-Xiang,Zhang, Gen-Xi,Wang, Jin-Yu,Li, Yuan,Zhang, Li-Jun,Wei, Yue,Wang, Hui-Hua,Zhang, Li,Hou, Qi-Rui Asian Australasian Association of Animal Productio 2010 Animal Bioscience Vol.23 No.2

The level of genetic differentiation and genetic structure in a Chinese native chicken breed, Bian chicken, and two controlled chicken populations (Jinghai chicken and Youxi chicken in China) were analysed based on 29 microsatellite markers. A total of 166 distinct alleles were observed across the 3 breeds, and 32 of these alleles (19.3%) were unique to only 1 breed. Bian chicken carried the largest number of private alleles at 15 (46.9%), followed by the Jinghai chicken with 12 private alleles (37.5%). The average polymorphism information content (0.5168) and the average expected heterozygote frequency (0.5750) of the Bian chicken were the highest, and those of the Jinghai chicken were 0.4915 and 0.5505, respectively, which were the lowest. Among 29 microsatellite loci, there were 15 highly informative loci in Bian chicken, and the other 14 were reasonably informative loci. The highly informative loci in Jinghai chicken and Youxi chicken were 17 and 14 respectively. Significant deviations from the Hardy-Weinberg equilibrium were observed at several locus-breed combinations, showing a deficit of heterozygotes in many cases. As a whole, genetic differentiation among the breeds estimated by the fixation index (Fst) were at 6.7% (p<0.001). The heterozygote deficit within population (Fis) was 22.2% (p<0.001), with the highest (0.249) in Bian chicken and lowest (0.159) in Youxi chicken. These results serve as an initial step in the plan for genetic characterization and conservation of the Chinese chicken genetic resource of Bian, as well as Jinghai and Youxi chickens.

ppGalNAc T1 as a Potential Novel Marker for Human Bladder Cancer

Ding, Ming-Xia,Wang, Hai-Feng,Wang, Jian-Song,Zhan, Hui,Zuo, Yi-Gang,Yang, De-Lin,Liu, Jing-Yu,Wang, Wei,Ke, Chang-Xing,Yan, Ru-Ping Asian Pacific Journal of Cancer Prevention 2012 Asian Pacific journal of cancer prevention Vol.13 No.11

Objectives: To investigate the effect of glycopeptide-preferring polypeptide GalNAc transferase 1 (ppGalNAc T1 ) targeted RNA interference (RNAi) on the growth and migration of human bladder carcinoma EJ cells in vitro and in vivo. Methods: DNA microarray assays were performed to determine ppGalNAc Ts(ppGalNAc T1-9) expression in human bladder cancer and normal bladder tissues. We transfected the EJ bladder cancer cell line with well-designed ppGalNAc T1 siRNA. Boyden chamber and Wound healing assays were used to investigate changes of shppGalNAc T1-EJ cell migration. Proliferation of shppGalNAc T1-EJ cells in vitro was assessed using [3H]-thymidine incorporation assay and soft agar colony formation assays. Subcutaneous bladder tumors in BALB/c nude mice were induced by inoculation of shppGalNAc T1-EJ cells and after inoculation diameters of tumors were measured every 5 days to determine gross tumor volumes. Results: ppGalNAc T1 mRNA in bladder cancer tissues was 11.2-fold higher than in normal bladder tissues. When ppGalNAc T1 expression in EJ cells was knocked down through transfection by pSUPER-shppGalNAc T1 vector, markedly reduced incorporation of [3H]-thymidine into DNA of EJ cells was observed at all time points compared with the empty vector transfected control cells. However, ppGalNAc T1 knockdown did not significantly inhibited cell migration (only 12.3%). Silenced ppGalNAc T1 expression significantly inhibited subcutaneous tumor growth compared with the control groups injected with empty vector transfected control cells. At the end of observation course (40 days), the inhibitory rate of cancerous growth for ppGalNAc T1 knockdown was 52.5%. Conclusion: ppGalNAc T1 might be a potential novel marker for human bladder cancer. Although ppGalNAc T1 knockdown caused no remarkable change in cell migration, silenced expression significantly inhibited proliferation and tumor growth of the bladder cancer EJ cell line.