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Small molecule natural compound agonist of SIRT3 as a therapeutic target for the treatment of intervertebral disc degeneration

  • Wang, Jianle;Nisar, Majid;Huang, Chongan;Pan, Xiangxiang;Lin, Dongdong;Zheng, Gang;Jin, Haiming;Chen, Deheng;Tian, Naifeng;Huang, Qianyu;Duan, Yue;Yan, Yingzhao;Wang, Ke;Wu, Congcong;Hu, Jianing;Zhang, Xiaolei;Wang, Xiangyang
    • Experimental and Molecular Medicine
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    • v.50 no.11
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    • pp.5.1-5.14
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    • 2018
  • Oxidative stress-induced mitochondrial dysfunction is implicated in the pathogenesis of intervertebral disc degeneration (IVDD). Sirtuin 3 (SIRT3), a sirtuin family protein located in mitochondria, is essential for mitochondrial homeostasis; however, the role of SIRT3 in the process of IVDD has remained elusive. Here, we explored the expression of SIRT3 in IVDD in vivo and in vitro; we also explored the role of SIRT3 in senescence, apoptosis, and mitochondrial homeostasis under oxidative stress. We subsequently activated SIRT3 using honokiol to evaluate its therapeutic potential for IVDD. We assessed SIRT3 expression in degenerative nucleus pulposus (NP) tissues and oxidative stress-induced nucleus pulposus cells (NPCs). SIRT3 was knocked down by lentivirus and activated by honokiol to determine its role in oxidative stress-induced NPCs. The mechanism by which honokiol affected SIRT3 regulation was investigated in vitro, and the therapeutic potential of honokiol was assessed in vitro and in vivo. We found that the expression of SIRT3 decreased with IVDD, and SIRT3 knockdown reduced the tolerance of NPCs to oxidative stress. Honokiol ($10{\mu}M$) improved the viability of NPCs under oxidative stress and promoted their properties of anti-oxidation, mitochondrial dynamics and mitophagy in a SIRT3-dependent manner. Furthermore, honokiol activated SIRT3 through the AMPK-PGC-$1{\alpha}$ signaling pathway. Moreover, honokiol treatment ameliorated IVDD in rats. Our study indicated that SIRT3 is involved in IVDD and showed the potential of the SIRT3 agonist honokiol for the treatment of IVDD.

Microstructure evolution and effect on deuterium retention in oxide dispersion strengthened tungsten during He+ irradiation

  • Ding, Xiao-Yu;Xu, Qiu;Zhu, Xiao-yong;Luo, Lai-Ma;Huang, Jian-Jun;Yu, Bin;Gao, Xiang;Li, Jian-Gang;Wu, Yu-Cheng
    • Nuclear Engineering and Technology
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    • v.52 no.12
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    • pp.2860-2866
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    • 2020
  • Oxide dispersion-strengthened materials W-1wt%Pr2O3 and W-1wt%La2O3 were synthesized by wet chemical method and spark plasma sintering. The field emission scanning electron microscopy (FE-SEM) analysis, XRD and Vickers microhardness measurements were conducted to characterize the samples. The irradiations were carried out with a 5 keV helium ion beam to fluences up to 5.0 × 1021 ions/m2 under 600 ℃ using the low-energy ion irradiation system. Transmission electron microscopy (TEM) study was performed to investigate the microstructural evolution in W-1wt%Pr2O3 and W-1wt%La2O3. At 1.0 × 1020 He+/m2, the average loops size of the W-1wt%Pr2O3 was 4.3 nm, much lower than W-1wt% La2O3 of 8.5 nm. However, helium bubbles were not observed throughout in both doped W materials. The effects of pre-irradiation with 1.0 × 1021 He+/m2 on trapping of injected deuterium in doped W was studied by thermal desorption spectrometry (TDS) technique using quadrupole mass spectrometer. Compared with the samples without He+ pre-irradiation, deuterium (D) retention of doped W materials increased after He+ irradiation, whose retention was unsaturated at the damage level of 1.0 × 1022D2+/m2. The present results implied that irradiation effect of He+ ions must be taken into account to evaluate the deuterium retention in fusion material applications.

Isolation and identification of goose skeletal muscle satellite cells and preliminary study on the function of C1q and tumor necrosis factor-related protein 3 gene

  • Wang, Han;He, Ke;Zeng, Xuehua;Zhou, Xiaolong;Yan, Feifei;Yang, Songbai;Zhao, Ayong
    • Animal Bioscience
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    • v.34 no.6
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    • pp.1078-1087
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    • 2021
  • Objective: Skeletal muscle satellite cells (SMSCs) are significant for the growth, regeneration, and maintenance of skeletal muscle after birth. However, currently, few studies have been performed on the isolation, culture and inducing differentiation of goose muscle satellite cells. Previous studies have shown that C1q and tumor necrosis factor-related protein 3 (CTRP3) participated in the process of muscle growth and development, but its role in the goose skeletal muscle development is not yet clear. This study aimed to isolate, culture, and identify the goose SMSCs in vitro. Additionally, to explore the function of CTRP3 in goose SMSCs. Methods: Goose SMSCs were isolated using 0.25% trypsin from leg muscle (LM) of 15 to 20 day fertilized goose eggs. Cell differentiation was induced by transferring the cells to differentiation medium with 2% horse serum and 1% penicillin streptomycin. Immunofluorescence staining of Desmin and Pax7 was used to identify goose SMSCs. Quantitative realtime polymerase chain reaction and western blot were applied to explore developmental expression profile of CTRP3 in LM and the regulation of CTRP3 on myosin heavy chains (MyHC), myogenin (MyoG) expression and Notch signaling pathway related genes expression. Results: The goose SMSCs were successfully isolated and cultured. The expression of Pax7 and Desmin were observed in the isolated cells. The expression of CTRP3 decreased significantly during leg muscle development. Overexpression of CTRP3 could enhance the expression of two myogenic differentiation marker genes, MyHC and MyoG. But knockdown of CTRP3 suppressed their expression. Furthermore, CTRP3 could repress the mRNA level of Notch signaling pathway-related genes, notch receptor 1, notch receptor 2 and hairy/enhancer-of-split related with YRPW motif 1, which previously showed a negative regulation in myoblast differentiation. Conclusion: These findings provide a useful cell model for the future research on goose muscle development and suggest that CTRP3 may play an essential role in skeletal muscle growth of goose.

Wind load and wind-induced effect of the large wind turbine tower-blade system considering blade yaw and interference

  • Ke, S.T.;Wang, X.H.;Ge, Y.J.
    • Wind and Structures
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    • v.28 no.2
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    • pp.71-87
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    • 2019
  • The yaw and interference effects of blades affect aerodynamic performance of large wind turbine system significantly, thus influencing wind-induced response and stability performance of the tower-blade system. In this study, the 5MW wind turbine which was developed by Nanjing University of Aeronautics and Astronautics (NUAA) was chosen as the research object. Large eddy simulation on flow field and aerodynamics of its wind turbine system with different yaw angles($0^{\circ}$, $5^{\circ}$, $10^{\circ}$, $20^{\circ}$, $30^{\circ}$ and $45^{\circ}$) under the most unfavorable blade position was carried out. Results were compared with codes and measurement results at home and abroad, which verified validity of large eddy simulation. On this basis, effects of yaw angle on average wind pressure, fluctuating wind pressure, lift coefficient, resistance coefficient,streaming and wake characteristics on different interference zone of tower of wind turbine were analyzed. Next, the blade-cabin-tower-foundation integrated coupling model of the large wind turbine was constructed based on finite element method. Dynamic characteristics, wind-induced response and stability performance of the wind turbine structural system under different yaw angle were analyzed systematically. Research results demonstrate that with the increase of yaw angle, the maximum negative pressure and extreme negative pressure of the significant interference zone of the tower present a V-shaped variation trend, whereas the layer resistance coefficient increases gradually. By contrast, the maximum negative pressure, extreme negative pressure and layer resistance coefficient of the non-interference zone remain basically same. Effects of streaming and wake weaken gradually. When the yaw angle increases to $45^{\circ}$, aerodynamic force of the tower is close with that when there's no blade yaw and interference. As the height of significant interference zone increases, layer resistance coefficient decreases firstly and then increases under different yaw angles. Maximum means and mean square error (MSE) of radial displacement under different yaw angles all occur at circumferential $0^{\circ}$ and $180^{\circ}$ of the tower. The maximum bending moment at tower bottom is at circumferential $20^{\circ}$. When the yaw angle is $0^{\circ}$, the maximum downwind displacement responses of different blades are higher than 2.7 m. With the increase of yaw angle, MSEs of radial displacement at tower top, downwind displacement of blades, internal force at blade roots all decrease gradually, while the critical wind speed decreases firstly and then increases and finally decreases. The comprehensive analysis shows that the worst aerodynamic performance and wind-induced response of the wind turbine system are achieved when the yaw angle is $0^{\circ}$, whereas the worst stability performance and ultimate bearing capacity are achieved when the yaw angle is $45^{\circ}$.

Change of PET Image According to CT Exposure Conditions (CT 촬영 조건에 따른 PET 영상의 변화)

  • Park, Jae-Yoon;Kim, Jung-hoon;Lee, Yong-Ki
    • Journal of the Korean Society of Radiology
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    • v.13 no.3
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    • pp.473-479
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    • 2019
  • PET-CT improves performance and reduces the time by combining PET and CT of spatial resolution, and uses CT scan for attenuation correction. This study analyzed PET image evaluation. The condition of the tube voltage and current of CT will be changed using. Uniformity phantom and resolution phantom were injected with 37 MBq $^{18}F$ (fluorine ; 511 keV, half life - 109.7 min), respectively. PET-CT (Biograph, siemens, US) was used to perform emission scan (30 min) and penetration scan. And then the collected image data were reconstructed in OSEM-3D. The same ROI was set on the image data with a analyzer (Vinci 2.54, Germany) and profile was used to analyze and compare spatial resolution and image quality through FWHM and SI. Analyzing profile with pre-defined ROI in each phantom, PET image was not influenced by the change of tube voltage or exposure dose. However, CT image was influenced by tube voltage, but not by exposure dose. When tube voltage was fixed and exposure dose changed, exposure dose changed too, increasing dose value. When exposure dose was fixed at 150 mA and tube voltage was varied, the result was 10.56, 24.6 and 35.61 mGy in each variables (in resolution phantom). In this study, attenuation image showed no significant difference when exposure dose was changed. However, when exposure dose increased, the amount of dose that patient absorbed increased too, which indicates that CT exposure dose should be decreased to minimum to lower the exposure dose that patient absorbs. Therefore future study needs to discuss the conditions that could minimize exposure dose that gets absorbed by patient during PET-CT scan.

Statistical Analysis of Count Rate Data for On-line Seawater Radioactivity Monitoring

  • Lee, Dong-Myung;Cong, Binh Do;Lee, Jun-Ho;Yeo, In-Young;Kim, Cheol-Su
    • Journal of Radiation Protection and Research
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    • v.44 no.2
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    • pp.64-71
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    • 2019
  • Background: It is very difficult to distinguish between a radioactive contamination source and background radiation from natural radionuclides in the marine environment by means of online monitoring system. The objective of this study was to investigate a statistical process for triggering abnormal level of count rate data measured from our on-line seawater radioactivity monitoring. Materials and Methods: Count rate data sets in time series were collected from 9 monitoring posts. All of the count rate data were measured every 15 minutes from the region of interest (ROI) for $^{137}Cs$ ($E_{\gamma}=661.6keV$) on the gamma-ray energy spectrum. The Shewhart ($3{\sigma}$), CUSUM, and Bayesian S-R control chart methods were evaluated and the comparative analysis of determination methods for count rate data was carried out in terms of the false positive incidence rate. All statistical algorithms were developed using R Programming by the authors. Results and Discussion: The $3{\sigma}$, CUSUM, and S-R analyses resulted in the average false positive incidence rate of $0.164{\pm}0.047%$, $0.064{\pm}0.0367%$, and $0.030{\pm}0.018%$, respectively. The S-R method has a lower value than that of the $3{\sigma}$ and CUSUM method, because the Bayesian S-R method use the information to evaluate a posterior distribution, even though the CUSUM control chart accumulate information from recent data points. As the result of comparison between net count rate and gross count rate measured in time series all the year at a monitoring post using the $3{\sigma}$ control charts, the two methods resulted in the false positive incidence rate of 0.142% and 0.219%, respectively. Conclusion: Bayesian S-R and CUSUM control charts are better suited for on-line seawater radioactivity monitoring with an count rate data in time series than $3{\sigma}$ control chart. However, it requires a continuous increasing trend to differentiate between a false positive and actual radioactive contamination. For the determination of count rate, the net count method is better than the gross count method because of relatively a small variation in the data points.

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.

Evaluation of the Biological Activity Affected by Extracting Solvents of Rosemary (Rosmarinus officinalis L.) (로즈마리(Rosmarinus officinalis L.) 극성별 용매 추출물의 생리활성 검증)

  • Li, Ke;Yang, Kyeong Hee;Guo, Lu;Cui, Zhengwei;Son, Beung Gu;Kang, Jum Soon;Lee, Yong Jae;Park, Young Hoon;Je, Beong Il;Choi, Young Whan
    • Journal of Life Science
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    • v.29 no.1
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    • pp.69-75
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    • 2019
  • Rosemary (Rosmarinus officinalis L.) is widely used as a food material. Although various physiological activities of rosemary have been reported, there have been no studies on the physiological activity of solvent extracts with different polarities. Rosemary extracts were obtained by extraction of dried powder using 0%, 25%, 50%, 70%, and 95% ethanol (EtOH) in distilled water, methanol, ethyl acetate, and hexane. As these ratios of EtOH are generally chosen by default and scarcely optimized, we investigated the impact of the composition of EtOH in distilled water on extract-related characteristics, such as DPPH free radical scavenging and ${\alpha}$-glucosidase inhibition, on the differentiation of 3T3-L1 adipocytes and inhibition of tyrosinase. Adipogenesis inhibition was highest at 70% EtOH. DPPH scavenging activity and inhibition of tyrosinase activity were reduced with 50% EtOH in water. However, inhibition of ${\alpha}$-glucosidase activity was higher in 50% EtOH in water. The best solvents in terms of DPPH scavenging activity, inhibition of tyrosinase and ${\alpha}$-glucosidase, and differentiation of adipocytes obtained with different concentrations of EtOH, although a lower similar activities were found with 50% ethanol. Considering the extraction solvents, a ratio of EtOH in water gives different content and constituents of compounds. These differences will give activities inhibition of adipogenesis, tyrosinase, ${\alpha}$-glucosidase activity, and DPPH scavenging activity.

Comparative study of thermal gelation properties and molecular forces of actomyosin extracted from normal and pale, soft and exudative-like chicken breast meat

  • Li, Ke;Liu, Jun-Ya;Fu, Lei;Zhao, Ying-Ying;Bai, Yan-Hong
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.5
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    • pp.721-733
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    • 2019
  • Objective: The objectives of this study were to investigate the thermal gelation properties and molecular forces of actomyosin extracted from two classes of chicken breast meat qualities (normal and pale, soft and exudative [PSE]-like) during heating process to further improve the understanding of the variations of functional properties between normal and PSE-like chicken breast meat. Methods: Actomyosin was extracted from normal and PSE-like chicken breast meat and the gel strength, water-holding capacity (WHC), protein loss, particle size and distribution, dynamic rheology and protein thermal stability were determined, then turbidity, active sulfhydryl group contents, hydrophobicity and molecular forces during thermal-induced gelling formation were comparatively studied. Results: Sodium dodecyl sulphate-polyacrylamide gel electrophoresis showed that protein profiles of actomyosin extracted from normal and PSE-like meat were not significantly different (p>0.05). Compared with normal actomyosin, PSE-like actomyosin had lower gel strength, WHC, particle size, less protein content involved in thermal gelation forming (p<0.05), and reduced onset temperature ($T_o$), thermal transition temperature ($T_d$), storage modulus (G') and loss modulus (G"). The turbidity, reactive sulfhydryl group of PSE-like actomyosin were higher when heated from $40^{\circ}C$ to $60^{\circ}C$. Further heating to $80^{\circ}C$ had lower transition from reactive sulfhydryl group into a disulfide bond and surface hydrophobicity. Molecular forces showed that hydrophobic interaction was the main force for heat-induced gel formation while both ionic and hydrogen bonds were different significantly between normal and PSE-like actomyosin (p<0.05). Conclusion: These changes in chemical groups and inter-molecular bonds affected protein-protein interaction and protein-water interaction and contributed to the inferior thermal gelation properties of PSE-like meat.

Profiles of Bacillus spp. Isolated from the Rhizosphere of Suaeda glauca and Their Potential to Promote Plant Growth and Suppress Fungal Phytopathogens

  • Lu, Ping;Jiang, Ke;Hao, Ya-Qiao;Chu, Wan-Ying;Xu, Yu-Dong;Yang, Jia-Yao;Chen, Jia-Le;Zeng, Guo-Hong;Gu, Zhou-Hang;Zhao, Hong-Xin
    • Journal of Microbiology and Biotechnology
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    • v.31 no.9
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    • pp.1231-1240
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    • 2021
  • Members of the genus Bacillus are known to play an important role in promoting plant growth and protecting plants against phytopathogenic microorganisms. In this study, 21 isolates of Bacillus spp. were obtained from the root micro-ecosystem of Suaeda glauca. Analysis of the 16S rRNA genes indicated that the isolates belong to the species Bacillus amyloliquefaciens, Bacillus velezensis, Bacillus subtilis, Bacillus pumilus, Bacillus aryabhattai and Brevibacterium frigoritolerans. One of the interesting findings of this study is that the four strains B1, B5, B16 and B21 are dominant in rhizosphere soil. Based on gyrA, gyrB, and rpoB gene analyses, B1, B5, and B21 were identified as B. amyloliquefaciens and B16 was identified as B. velezensis. Estimation of antifungal activity showed that the isolate B1 had a significant inhibitory effect on Fusarium verticillioides, B5 and B16 on Colletotrichum capsici (syd.) Butl, and B21 on Rhizoctonia cerealis van der Hoeven. The four strains grew well in medium with 1-10% NaCl, a pH value of 5-8, and promoted the growth of Arabidopsis thaliana. Our results indicate that these strains may be promising agents for the biocontrol and promotion of plant growth and further study of the relevant bacteria will provide a useful reference for the development of microbial resources.