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Inhibition of MicroRNA-15a/16 Expression Alleviates Neuropathic Pain Development through Upregulation of G Protein-Coupled Receptor Kinase 2

  • Li, Tao;Wan, Yingchun;Sun, Lijuan;Tao, Shoujun;Chen, Peng;Liu, Caihua;Wang, Ke;Zhou, Changyu;Zhao, Guoqing
    • Biomolecules & Therapeutics
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    • v.27 no.4
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    • pp.414-422
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    • 2019
  • There is accumulating evidence that microRNAs are emerging as pivotal regulators in the development and progression of neuropathic pain. MicroRNA-15a/16 (miR-15a/16) have been reported to play an important role in various diseases and inflammation response processes. However, whether miR-15a/16 participates in the regulation of neuroinflammation and neuropathic pain development remains unknown. In this study, we established a mouse model of neuropathic pain by chronic constriction injury (CCI) of the sciatic nerves. Our results showed that both miR-15a and miR-16 expression was significantly upregulated in the spinal cord of CCI rats. Downregulation of the expression of miR-15a and miR-16 by intrathecal injection of a specific inhibitor significantly attenuated the mechanical allodynia and thermal hyperalgesia of CCI rats. Furthermore, inhibition of miR-15a and miR-16 downregulated the expression of interleukin-$1{\beta}$ and tumor-necrosis factor-${\alpha}$ in the spinal cord of CCI rats. Bioinformatic analysis predicted that G protein-coupled receptor kinase 2 (GRK2), an important regulator in neuropathic pain and inflammation, was a potential target gene of miR-15a and miR-16. Inhibition of miR-15a and miR-16 markedly increased the expression of GRK2 while downregulating the activation of p38 mitogen-activated protein kinase and $NF-{\kappa}B$ in CCI rats. Notably, the silencing of GRK2 significantly reversed the inhibitory effects of miR-15a/16 inhibition in neuropathic pain. In conclusion, our results suggest that inhibition of miR-15a/16 expression alleviates neuropathic pain development by targeting GRK2. These findings provide novel insights into the molecular pathogenesis of neuropathic pain and suggest potential therapeutic targets for preventing neuropathic pain development.

Association analysis of polymorphisms of G protein-coupled receptor 54 gene exons with reproductive traits in Jiaxing Black sows

  • Wu, Fen;Zhang, Wei;Song, Qian-Qian;Li, Hai-Hong;Xu, Ming-Shu;Liu, Guo-Liang;Zhang, Jin-Zhi
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.8
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    • pp.1104-1111
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    • 2019
  • Objective: The aim of this study was to detect single nucleotide polymorphisms (SNP) of G protein-coupled receptor 54 (GPR54) gene and explore association of this candidate gene with reproductive traits in Jiaxing Black sows. Methods: Six pairs of primers of the gene were designed to amplify all exons thus sequences of which were detected by means of direct sequencing and then SNP loci were scanned. The effects of SNPs on total number of piglets born (TNB), number of piglets born alive (NBA), number of still born piglets (NSB), and litter weight at birth (LWB) of Jiaxing Black sows were analyzed. Results: Three SNP loci, including T3739C, C3878T and T6789C, were identified via comparison of sequencing and two genotypes (AB, BB) at each SNP site were observed. T3739C resulted in the change of amino acid ($Leu{\rightarrow}Pro$) in corresponding protein, and C3878T resulted in synonymous mutation ($Ile{\rightarrow}Ile$). Statistical results demonstrated that allele B was the preponderant allele at the three SNP loci and Genotype BB was the preponderant genotype. Meanwhile, Chi-Square test of these three SNPs indicated that all mutation sites fitted in Hardy-Weinberg equilibrium (p>0.05). For GPR54-T3739C locus, Jiaxing Black sows with genotype BB had 1.23 TNB and 1.28 NBA (p<0.01) that were more than those with genotype AB, respectively. Jiaxing Black sows that had the first two parities with genotype BB had additional 2.23 TNB, 2.27 NBA (p<0.01), and 1.94 LWB (p<0.05) compared to those with genotype AB, respectively. However, for other two loci, no significant difference was found between TNB, NBA, NSB, and LWB, and different genotypes of Jiaxing Black sows. Conclusion: In conclusion, the polymorphisms of GPR54-T3739C locus were significantly associated to TNB, NBA, and LWB and could be used as a potential genetic marker to improve reproductive function of Jiaxing black sows.

Development of LiDAR-Based MRM Algorithm for LKS System (LKS 시스템을 위한 라이다 기반 MRM 알고리즘 개발)

  • Son, Weon Il;Oh, Tae Young;Park, Kihong
    • The Journal of The Korea Institute of Intelligent Transport Systems
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    • v.20 no.1
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    • pp.174-192
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    • 2021
  • The LIDAR sensor, which provides higher cognitive performance than cameras and radar, is difficult to apply to ADAS or autonomous driving because of its high price. On the other hand, as the price is decreasing rapidly, expectations are rising to improve existing autonomous driving functions by taking advantage of the LIDAR sensor. In level 3 autonomous vehicles, when a dangerous situation in the cognitive module occurs due to a sensor defect or sensor limit, the driver must take control of the vehicle for manual driving. If the driver does not respond to the request, the system must automatically kick in and implement a minimum risk maneuver to maintain the risk within a tolerable level. In this study, based on this background, a LIDAR-based LKS MRM algorithm was developed for the case when the normal operation of LKS was not possible due to troubles in the cognitive system. From point cloud data collected by LIDAR, the algorithm generates the trajectory of the vehicle in front through object clustering and converts it to the target waypoints of its own. Hence, if the camera-based LKS is not operating normally, LIDAR-based path tracking control is performed as MRM. The HAZOP method was used to identify the risk sources in the LKS cognitive systems. B, and based on this, test scenarios were derived and used in the validation process by simulation. The simulation results indicated that the LIDAR-based LKS MRM algorithm of this study prevents lane departure in dangerous situations caused by various problems or difficulties in the LKS cognitive systems and could prevent possible traffic accidents.

Acetylation of Sarcoplasmic and Myofibrillar Proteins were Associated with Ovine Meat Quality Attributes at Early Postmortem

  • Zhang, Yejun;Li, Xin;Zhang, Dequan;Ren, Chi;Bai, Yuqiang;Ijaz, Muawuz;Wang, Xu;Zhao, Yingxin
    • Food Science of Animal Resources
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    • v.41 no.4
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    • pp.650-663
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    • 2021
  • The objective of this study was to examine the relationship between meat quality attributes and the changes of sarcoplasmic protein acetylation and myofibrillar protein acetylation in lamb longissimus thoracis et lumborum muscles at different postmortem phases. Protein acetylation, color, pH, shear force, myofibril fragmentation index and cooking loss were measured. The total level of acetylated sarcoplasmic proteins showed a negative relation with pH, a positive relation with a*, b* and cooking loss at the pre-rigor phase. Sarcoplasmic proteins acetylation affected postmortem pH by regulating glycolysis, which in turn affects color and cooking loss. The total level of acetylated myofibrillar proteins showed a positive relation with shear force at the pre-rigor phase. Myofibrillar proteins acetylation affected meat tenderness by regulating muscle contraction. This study indicated that acetylation played a regulatory role of meat color, water-holding capacity, and tenderization process at early postmortem.

Effect of Postmortem Phases on Lamb Meat Quality: A Physicochemical, Microstructural and Water Mobility Approach

  • Ge, Yue;Zhang, Dequan;Zhang, Huimin;Li, Xin;Fang, Fei;Liang, Ce;Wang, Zhenyu
    • Food Science of Animal Resources
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    • v.41 no.5
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    • pp.802-815
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    • 2021
  • To investigate the effect of postmortem phases on lamb meat quality, the physicochemical quality, microstructure and water mobility of oyster cut, short loin, knuckle and silverside muscles from Small-Tail Han sheep were evaluated in the pre-rigor, rigor mortis and post-rigor phases. Pre-rigor lamb meat had higher pH and water holding capacity (WHC), whereas lower CIE L*, b*, hue angle values than rigor mortis and post-rigor meat (p<0.05). The Warner-Bratzler shear force (WBSF) values were higher in rigor mortis short loin and silverside than their pre-rigor and post-rigor counterparts, pre-rigor short loin had lower WBSF value than its post-rigor counterpart (p<0.05). Muscle fibers shrank laterally and longitudinally during the onset of rigor mortis. Rigor mortis and postrigor lamb meat exhibited wide I-bands, dark A-bands, short sarcomeres and large inter-myofibrillar spaces. The shift of immobilized water to free water and repulsion from the intra-myofibrillar space to the extracellular space result in the increase of water loss in rigor mortis and post-rigor lamb meat. The results of the principal component analysis (PCA) indicated that rigor mortis and post-rigor lamb meat had similar quality properties but different from pre-rigor lamb meat. In conclusion, the lamb meat in the pre-rigor phase had good tenderness, color and WHC. The results of this research could provide some theoretical references for lamb meat production and processing.

Effects of dietary supplementation of glucose oxidase, catalase, or both on reproductive performance, oxidative stress, fecal microflora and apoptosis in multiparous sows

  • Sun, Xiaojiao;Piao, Longguo;Jin, Haifeng;Nogoy, K. Margarette C.;Zhang, Junfang;Sun, Bin;Jin, Yi;Lee, Dong Hoon;Choi, Seong-Ho;Smith, Stephen B;Li, Xiangzi
    • Animal Bioscience
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    • v.35 no.1
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    • pp.75-86
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    • 2022
  • Objective: The objective of this experiment was to investigate the effect of dietary glucose oxidase (GOD), catalase (CAT), or both supplementation on reproductive performance, oxidative stress, and apoptosis in sows. Methods: A total of 104 multiparous sows were randomly assigned to four groups (n = 26) with each group given a basal diet, basal diet plus GOD at 60 U/kg, basal diet plus CAT at 75 U/kg, and basal diet plus GOD at 60 U/kg and CAT at 75 U/kg. Sows were fed the experimental diets throughout gestation and lactation. Results: Dietary GOD supplementation increased average daily feed intake of sows and litter weight at weaning (p<0.05). Dietary CAT supplementation reduced the duration of parturition, stillbirth, and piglet mortality and increased growth performance of weaned piglets (p<0.05). Dietary GOD and CAT supplementation enhanced antioxidant enzyme activities and lessened oxidative stress product levels in plasma of sows and elevated antioxidant capacity of 14-day milk and plasma in weaned piglets (p<0.05). Dietary GOD supplementation increased fecal Lactobacillus counts and reduced Escherichia coli counts of sows (p<0.05). Compared with the basal diet, the GOD diet reduced fecal Escherichia coli counts of sows, but the addition of CAT did not reduce Escherichia coli counts in the GOD diet. Dietary GOD and CAT supplementation reduced the apoptosis rate of the liver, endometrium, and ovarian granulosa cells in sows (p<0.05). In the liver, uterus, and ovary of sows, the mRNA expression of caspase-3 and caspase-9 was downregulated by dietary GOD and CAT supplementation (p<0.05). Conclusion: Dietary GOD and CAT supplementation could improve the antioxidant capacity of sows and weaned piglets, and alleviate hepatic, ovarian and uterine apoptosis by weakening apoptosis-related gene expression. Glucose oxidase regulated fecal microflora of sows, but supplementation of CAT to GOD could weaken the inhibitory effect of GOD on fecal Escherichia coli.

Effects of exploration and molecular mechanism of CsV on eNOS and vascular endothelial functions

  • Zuo, Deyu;Jiang, Heng;Yi, Shixiong;Fu, Yang;Xie, Lei;Peng, Qifeng;Liu, Pei;Zhou, Jie;Li, Xunjia
    • Advances in nano research
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    • v.12 no.5
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    • pp.501-514
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    • 2022
  • This study aimed to investigate the effects and potential mechanisms of Chikusetsusaponin V (CsV) on endothelial nitric oxide synthase (eNOS) and vascular endothelial cell functions. Different concentrations of CsV were added to animal models, bovine aorta endothelial cells (BAECs) and human umbilical vein endothelial cells (HUVECs) cultured in vitro. qPCR, Western blotting (WB), and B ultrasound were performed to explore the effects of CsV on mouse endothelial cell functions, vascular stiffness and cellular eNOS mRNA, protein expression and NO release. Bioinformatics analysis, network pharmacology, molecular docking and protein mass spectrometry analysis were conducted to jointly predict the upstream transcription factors of eNOS. Furthermore, pulldown and ChIP and dual luciferase assays were employed for subsequent verification. At the presence or absence of CsV stimulation, either overexpression or knockdown of purine rich element binding protein A (PURA) was conducted, and PCR assay was employed to detect PURA and eNOS mRNA expressions, Western blot was used to detect PURA and eNOS protein expressions, cell NO release and serum NO levels. Tube formation experiment was conducted to detect the tube forming capability of HUVECs cells. The animal vasodilation function test detected the vasodilation functions. Ultrasonic detection was performed to determine the mouse aortic arch pulse wave velocity to identify aortic stiffness. CsV stimulus on bovine aortic cells revealed that CsV could upregulate eNOS protein levels in vascular endothelial cells in a concentration and time dependent manner. The expression levels of eNOS mRNA and phosphorylation sites Ser1177, Ser633 and Thr495 increased significantly after CsV stimulation. Meanwhile, CsV could also enhance the tube forming capability of HUVECs cells. Following the mice were gavaged using CsV, the eNOS protein level of mouse aortic endothelial cells was upregulated in a concentration- and time-dependent manner, and serum NO release and vasodilation ability were simultaneously elevated whereas arterial stiffness was alleviated. The pulldown, ChIP and dual luciferase assays demonstrated that PURA could bind to the eNOS promoter and facilitate the transcription of eNOS. Under the conditions of presence or absence of CsV stimulation, overexpression or knockdown of PURA indicated that the effect of CsV on vascular endothelial function and eNOS was weakened following PURA gene silence, whereas overexpression of PURA gene could enhance the effect of CsV upregulating eNOS expression. CsV could promote NO release from endothelial cells by upregulating the expression of PURA/eNOS pathway, improve endothelial cell functions, enhance vasodilation capability, and alleviate vessel stiffness. The present study plays a role in offering a theoretical basis for the development and application of CsV in vascular function improvement, and it also provides a more comprehensive understanding of the pharmacodynamics of CsV.

Effects of Protein Intake on Sarcopenia Prevention and Physical Function of the Elderly in a Rural Community of South Korea During the COVID-19 Pandemic (COVID-19시대 한국 농촌 지역 노인의 단백질 섭취가 근감소증 예방 및 신체기능에 미치는 영향)

  • Li, Xinxing;Kim, Hee-Jae;Kim, Dae-Young;Zhang, Yanjie;Seo, Ji-Won;Ahn, Seo-Hyun;Song, Wook
    • Journal of The Korean Society of Integrative Medicine
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    • v.10 no.2
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    • pp.37-47
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    • 2022
  • Purpose : We aimed to investigate the effects of nutritional foods on sarcopenia prevention and physical function among the elderly living in rural communities during the COVID-19 pandemic by providing customized nutrition. Methods : This study was conducted in the rural community of SCC. Participants (n=24, over age 65) were randomly assigned into a Protein group (n=12) and a Vitamin group (n=12). The protein group was given 23 g/d of protein (whey, soybean, BCAA) for 8 weeks and the Vitamin group 23 g/d of vitamin (B, C, D, E and mixed minerals such as calcium, magnesium, zinc) for 8 weeks. All participants had their body composition such as height, weight, skeletal muscle mass, body mass index, and body fat percentage, measured using bioelectrical impedance analysis (BIA) and physical function assessed using grip strength and the short physical performance battery (SPPB). Results : At the end of the intervention, there was a significant increase in skeletal muscle mass (p<.01) in the Protein group (p=.002, 4.92 %) compared to the baseline: it increased by 2.33 %. The Vitamin group had a significant increase in body fat percentage after the intervention (p=.001, 15.35 %) compared to the baseline: body fat percentage decreased by 4.49 %. There were no significant differences in left and right Grip strength/Weight, SPPB, 4-m gait speed, chair stand test, and sense of balance in both groups. Conclusion : The findings from this study suggest that 8 weeks of protein intake have a significant effect on skeletal muscle mass and body fat percentage. Protein intake helped promoting the health of the elderly in rural community during the COVID-19 pandemic. It will assist creating a foundation for providing customized nutrition for the elderly in rural community in the future.

Epidemiological investigation of porcine pseudorabies virus and its coinfection rate in Shandong Province in China from 2015 to 2018

  • Ma, Zicheng;Han, Zifeng;Liu, Zhaohu;Meng, Fanliang;Wang, Hongyu;Cao, Longlong;Li, Yan;Jiao, Qiulin;Liu, Sidang;Liu, Mengda
    • Journal of Veterinary Science
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    • v.21 no.3
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    • pp.36.1-36.9
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    • 2020
  • Background: Pseudorabies, also known as Aujeszky's disease, is caused by the pseudorabies virus (PRV) and has been recognized as a critical disease affecting the pig industry and a wide range of animals around the world, resulting in great economic losses each year. Shandong province, one of the most vital food animal-breeding regions in China, has a very dense pig population, within which pseudorabies infections were detected in recent years. The data, however, on PRV epidemiology and coinfection rates of PRV with other major swine diseases is sparse. Objectives: This study aimed to investigate the PRV epidemiology in Shandong and analyze the current control measures. Methods: In this study, a total number of 16,457 serum samples and 1,638 tissue samples, which were collected from 362 intensive pig farms (≥ 300 sows/farm) covered all cities in Shandong, were tested by performing enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR). Results: Overall, 52.7% and 91.5% of the serum samples were positive for PRV-gE and -gB, respectively, based on ELISA results. In addition, 15.7% of the tissue samples were PCR positive for PRV. The coinfection rates of PRV with porcine circovirus type 2 (PCV2), porcine reproductive and respiratory syndrome virus, and classical swine fever virus were measured; coinfection with PCV2 was 35.0%, higher than those of the other two viruses. Macroscopic and microscopic lesions were observed in various tissues during histopathological examination. Conclusions: The results demonstrate the PRV prevalence and its coinfection rates in Shandong province and indicate that pseudorabies is endemic in pig farms in this region. This study provides epidemiological data that can be useful in the prevention and control of pseudorabies in Shandong, China.

Numerical Modeling of Thermoshearing in Critically Stressed Rough Rock Fracture: DECOVALEX-2023 Task G (임계응력 하 거친 암석 균열의 Thermoshearing 수치모델링: 국제공동연구 DECOVALEX-2023 Task G)

  • Jung-Wook Park;Chan-Hee Park;Li Zhuang;Jeoung Seok Yoon;Changlun Sun;Changsoo Lee
    • Tunnel and Underground Space
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    • v.33 no.3
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    • pp.189-207
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    • 2023
  • In the present study, the thermoshearing experiment on a rough rock fracture were modeled using a three-dimensional grain-based distinct element model (GBDEM). The experiment was conducted by the Korea Institute of Construction Technology to investigate the progressive shear failure of fracture under the influence of thermal stress in a critical stress state. The numerical model employs an assembly of multiple polyhedral grains and their interfaces to represent the rock sample, and calculates the coupled thermo-mechanical behavior of the grains (blocks) and the interfaces (contacts) using 3DEC, a DEM code. The primary focus was on simulating the temperature evolution, generation of thermal stress, and shear and normal displacements of the fracture. Two fracture models, namely the mated fracture model and the unmated fracture model, were constructed based on the degree of surface matedness, and their respective behaviors were compared and analyzed. By leveraging the advantage of the DEM, the contact area between the fracture surfaces was continuously monitored during the simulation, enabling an examination of its influence on shear behavior. The numerical results demonstrated distinct differences depending on the degree of the surface matedness at the initial stage. In the mated fracture model, where the surfaces were in almost full contact, the characteristic stages of peak stress and residual stress commonly observed in shear behavior of natural rock joints were reasonably replicated, despite exhibiting discrepancies with the experimental results. The analysis of contact area variation over time confirmed that our numerical model effectively simulated the abrupt normal dilation and shear slip, stress softening phenomenon, and transition to the residual state that occur during the peak stress stage. The unmated fracture model, which closely resembled the experimental specimen, showed qualitative agreement with the experimental observations, including heat transfer characteristics, the progressive shear failure process induced by heating, and the increase in thermal stress. However, there were some mismatches between the numerical and experimental results regarding the onset of fracture slip and the magnitudes of fracture stress and displacement. This research was conducted as part of DECOVALEX-2023 Task G, and we expect the numerical model to be enhanced through continued collaboration with other research teams and validated in further studies.