Please wait a minute...
 Home  About the Journal Editorial Board Aims & Scope Peer Review Policy Subscription Contact us
 
Early Edition  //  Current Issue  //  Open Special Issues  //  Archives  //  Most Read  //  Most Downloaded  //  Most Cited
Aging and disease    2021, Vol. 12 Issue (1) : 50-60     DOI: 10.14336/AD.2020.0623
Orginal Article |
Transcranial Focused Ultrasound Stimulation Improves Neurorehabilitation after Middle Cerebral Artery Occlusion in Mice
Jixian Wang1, Guofeng Li3,4, Lidong Deng2, Muyassar Mamtilahun2, Lu Jiang2, Weibao Qiu3, Hairong Zheng3, Junfeng Sun2, Qing Xie1,*, Guo-Yuan Yang2,*
1Department of Rehabilitation, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
2Med-X Research Institute and School of Biomedical Engineering, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
3Shenzhen Institutes of Advanced Technology Chinese Academy of Sciences, Shenzhen 518055, China.
4School of Information Engineering, Guangdong Medical University, Dongguan 523808, China
Download: PDF(577 KB)   HTML
Export: BibTeX | EndNote | Reference Manager | ProCite | RefWorks    
Abstract  

Transcranial focused ultrasound stimulation (tFUS) regulates neural activity in different brain regions in humans and animals. However, the role of ultrasound stimulation in modulating neural activity and promoting neurorehabilitation in the ischemic brain is largely unknown. In the present study, we explored the effect of tFUS on neurological rehabilitation and the underlying mechanism. Adult male ICR mice (n=42) underwent transient middle cerebral artery occlusion. One week after brain ischemia, low frequency (0.5 MHz) tFUS was applied to stimulate the ischemic hemisphere of mice for 7 consecutive days (10 minutes daily). Brain infarct volume, neurobehavioral tests, microglia activation, IL-10 and IL-10R levels were further assessed for up to 14 days. We found that the brain infarct volume was significantly reduced in the tFUS treated mice compared to that in the non-treated mice (p<0.05). Similarly, neurological severity scores, elevated body swing test, and corner test improved in the tFUS treated mice (p<0.05). We also demonstrated that tFUS resulted in increased M2 microglia in the ischemic brain region. The expression of IL-10R and IL-10 levels were also substantially upregulated (p<0.05). We concluded that tFUS served as a unique technique to promote neurorehabilitation after brain ischemia by promoting microglia polarization and further regulating IL-10 signaling in the ischemic brain.

Keywords ischemia      microglia      rehabilitation      stimulation      ultrasound     
Corresponding Authors: Xie Qing,Yang Guo-Yuan   
About author:

these authors contributed equally to this work.

Just Accepted Date: 02 July 2020   Issue Date: 11 January 2021
Service
E-mail this article
E-mail Alert
RSS
Articles by authors
Wang Jixian
Li Guofeng
Deng Lidong
Mamtilahun Muyassar
Jiang Lu
Qiu Weibao
Zheng Hairong
Sun Junfeng
Xie Qing
Yang Guo-Yuan
Cite this article:   
Wang Jixian,Li Guofeng,Deng Lidong, et al. Transcranial Focused Ultrasound Stimulation Improves Neurorehabilitation after Middle Cerebral Artery Occlusion in Mice[J]. Aging and disease, 2021, 12(1): 50-60.
URL:  
http://www.aginganddisease.org/EN/10.14336/AD.2020.0623     OR
Figure 1.  Flow diagram of the experimental design Animals underwent middle cerebral artery occlusion (MCAO). tFUS was applied for 7 days from day 7 of MCAO. The behavioral tests were performed following MCAO for 14 days. Brain atrophy volume and potential mechanisms were examined.
Figure 2.  The setup used for low frequency transcranial ultrasound stimulation of the ischemic hemisphere of mice A) Photograph showed the experimental setup of ultrasound stimulation device. B) Photograph showed the tFUS process. The transducer was located on the ipsilateral hemisphere of the mouse brain. C) The photograph showed the ultrasound intensity distribution field relative to the spatial peak in the sanitation region. The Z axis represented the stimulus depth. The Y axis showed the stimulus width. Different colors represented different relative tFUS intensities. Red (high intensity); violet (low intensity). D) The photograph showed a general view of the tFUS sequences: 0.5 ms tone-burst-duration (TBD), 1000 Hz pulse repetition frequency (PRF), 300 ms sonication duration (SD), and 2.7 s inter-stimuli interval (ISI). Green, red and blue arrows indicate the portable tFUS system, the impedance matching device and the ultrasound transducer, respectively.
Figure 3.  Ultrasound stimulation of the brain promoted neurofunctional recovery in mice that underwent MCAO A) Photomicrographs showed representative sets of cresyl violet-stained brain sections from mice treated with or without tFUS after 60 minutes of MCAO. The line illustrates the atrophy volume of the ipsilateral hemisphere following 7 days of tFUS treatment. Bar graph showed the semi-quantitative data from panel a. n=6 per group, p<0.05, tFUS treated mice after MCAO vs. MCAO alone. Line graphs showed the neurologic severity score (B), EBST (C) and corner test (D) results for the sham, MCAO alone, and MCAO mice with tFUS. n=6 per group, p<0.05, tFUS treated MCAO mice vs. MCAO alone group.
Figure 4.  Ultrasound stimulation of the brain increased the number of M2 microglia after MCAO A) Co-immunostaining of the M1 microglia markers CD16/32 and IBA-1 (upper left), and the M2 microglia markers arginase and IBA-1 (upper right) in the mouse brain after one hour of MCAO in mice. After tFUS, there were fewer CD16/32+/IBA-1+ cells were detected while arginase+/IBA-1+ cells were increased in the ischemic perifocal region (below). Bar=20 μm. B) Bar graph showed the number of M1 and M2 microglia in the ischemic mouse brain treated with tFUS. Data are presented as mean ± SD. ***, p<0.001. C) CD16/32/CX3CR1+ (M1) and CD206/CX3CR1+ (M2) microglia, the number of CD16/32/CX3CR1+and CD206/CX3CR1+ microglia were increased. However, the number of CD206/CX3CR1+ cells was increased remarkably in the tFUS treated group. D) Bar graph showed the number of M1 and M2 microglia in the ischemic mouse brain treated with tFUS. Data are presented as mean ± SD. *, p<0.05.
Figure 5.  Ultrasound stimulation of the brain upregulated IL-10R and IL-10 A) Photomicrographs showed IL-10R (green) and DAPI (blue) staining in the perifocal region of the ipsilateral hemisphere in sham, MCAO alone, and MCAO mice with tFUS. White arrowheads indicate that IL-10R is mainly expressed in the cell cytosol in the perifocal region in MCAO mice. IL-10R staining was greatly increased after tFUS. Scale bar= 25 μm. B) Western blot analysis showed that IL-10 expression (101 KD) was greatly increased after tFUS. The bar graph showed the quantitative data for the Western blot analysis. n=6 per group. *, p<0.05, tFUS treated mice after MCAO vs. MCAO alone. PCR showed the relative expression of IL-10 (C) and IL-10R (D) in the brain after tFUS treatment. n=6 per group. Data are presented as mean ± SD.*, p<0.05.
[1] Richards CL, Malouin F, Nadeau S (2015). Stroke rehabilitation: clinical picture, assessment, and therapeutic challenge. Prog Brain Res, 218:253-280.
[2] Klomjai W, Katz R, Lackmy-Vallee A (2015). Basic principles of transcranial magnetic stimulation (TMS) and repetitive TMS (rTMS). Ann Phys Rehabil Med, 58:208-213.
[3] Matsumoto H, Ugawa Y (2017). Adverse events of tDCS and tACS: A review. Clin Neurophysiol Pract, 2:19-25.
[4] Bergmann TO, Karabanov A, Hartwigsen G, Thielscher A, Siebner HR (2016). Combining non-invasive transcranial brain stimulation with neuroimaging and electrophysiology: Current approaches and future perspectives. Neuroimage, 140:4-19.
[5] Dayan E, Censor N, Buch ER, Sandrini M, Cohen LG (2013). Noninvasive brain stimulation: from physiology to network dynamics and back. Nat Neurosci, 16:838-844.
[6] Karabanov A, Ziemann U, Hamada M, George MS, Quartarone A, Classen J, et al. (2015). Consensus Paper: Probing Homeostatic Plasticity of Human Cortex With Non-invasive Transcranial Brain Stimulation. Brain Stimul, 8:993-1006.
[7] Ziemann U, Siebner HR (2008). Modifying motor learning through gating and homeostatic metaplasticity. Brain Stimul, 1:60-66.
[8] Herz DM, Christensen MS, Bruggemann N, Hulme OJ, Ridderinkhof KR, Madsen KH, et al. (2014). Motivational tuning of fronto-subthalamic connectivity facilitates control of action impulses. J Neurosci, 34:3210-3217.
[9] Lefaucheur JP, Aleman A, Baeken C, Benninger DH, Brunelin J, Di Lazzaro V, et al. (2020). Evidence-based guidelines on the therapeutic use of repetitive transcranial magnetic stimulation (rTMS): An update (2014-2018). Clin Neurophysiol, 131:474-528.
[10] Loo CK, Mitchell PB (2005). A review of the efficacy of transcranial magnetic stimulation (TMS) treatment for depression, and current and future strategies to optimize efficacy. J Affect Disord, 88:255-267.
[11] Mancuso JJ, Kim J, Lee S, Tsuda S, Chow NB, Augustine GJ (2011). Optogenetic probing of functional brain circuitry. Exp Physiol, 96:26-33.
[12] He X, Lu Y, Lin X, Jiang L, Tang Y, Tang G, et al. (2017). Optical inhibition of striatal neurons promotes focal neurogenesis and neurobehavioral recovery in mice after middle cerebral artery occlusion. J Cereb Blood Flow Metab, 37:837-847.
[13] Jiang L, Li W, Mamtilahun M, Song Y, Ma Y, Qu M, et al. (2017). Optogenetic Inhibition of Striatal GABAergic Neuronal Activity Improves Outcomes After Ischemic Brain Injury. Stroke, 48:3375-3383.
[14] Fini M, Tyler WJ (2017). Transcranial focused ultrasound: a new tool for non-invasive neuromodulation. Int Rev Psychiatry, 29:168-177.
[15] Naor O, Krupa S, Shoham S (2016). Ultrasonic neuromodulation. J Neural Eng, 13:031003.
[16] Tufail Y, Matyushov A, Baldwin N, Tauchmann ML, Georges J, Yoshihiro A, et al. (2010). Transcranial pulsed ultrasound stimulates intact brain circuits. Neuron, 66:681-694.
[17] King RL, Brown JR, Newsome WT, Pauly KB (2013). Effective parameters for ultrasound-induced in vivo neurostimulation. Ultrasound Med Biol, 39:312-331.
[18] Younan Y, Deffieux T, Larrat B, Fink M, Tanter M, Aubry JF (2013). Influence of the pressure field distribution in transcranial ultrasonic neurostimulation. Med Phys, 40:082902.
[19] Yoo SS, Bystritsky A, Lee JH, Zhang Y, Fischer K, Min BK, et al. (2011). Focused ultrasound modulates region-specific brain activity. Neuroimage, 56:1267-1275.
[20] Lee W, Lee SD, Park MY, Foley L, Purcell-Estabrook E, Kim H, et al. (2016). Image-Guided Focused Ultrasound-Mediated Regional Brain Stimulation in Sheep. Ultrasound Med Biol, 42:459-470.
[21] Dallapiazza RF, Timbie KF, Holmberg S, Gatesman J, Lopes MB, Price RJ, et al. (2018). Noninvasive neuromodulation and thalamic mapping with low-intensity focused ultrasound. J Neurosurg, 128:875-884.
[22] Deffieux T, Younan Y, Wattiez N, Tanter M, Pouget P, Aubry JF (2013). Low-intensity focused ultrasound modulates monkey visuomotor behavior. Curr Biol, 23:2430-2433.
[23] Legon W, Bansal P, Tyshynsky R, Ai L, Mueller JK (2018). Transcranial focused ultrasound neuromodulation of the human primary motor cortex. Sci Rep, 8:10007.
[24] Lee W, Kim H, Jung Y, Song IU, Chung YA, Yoo SS (2015). Image-guided transcranial focused ultrasound stimulates human primary somatosensory cortex. Sci Rep, 5:8743.
[25] Lee W, Chung YA, Jung Y, Song IU, Yoo SS (2016). Simultaneous acoustic stimulation of human primary and secondary somatosensory cortices using transcranial focused ultrasound. BMC Neurosci, 17:68.
[26] Lee W, Kim HC, Jung Y, Chung YA, Song IU, Lee JH, et al. (2016). Transcranial focused ultrasound stimulation of human primary visual cortex. Sci Rep, 6:34026.
[27] Kubanek J (2018). Neuromodulation with transcranial focused ultrasound. Neurosurg Focus, 44:E14.
[28] Lele PP (1962). A simple method for production of trackless focal lesions with focused ultrasound: physical factors. J Physiol, 160:494-512.
[29] McDannold NJ, Jolesz FA, Hynynen KH (1999). Determination of the optimal delay between sonications during focused ultrasound surgery in rabbits by using MR imaging to monitor thermal buildup in vivo. Radiology, 211:419-426.
[30] Clement GT, White PJ, King RL, McDannold N, Hynynen K (2005). A magnetic resonance imaging-compatible, large-scale array for trans-skull ultrasound surgery and therapy. J Ultrasound Med, 24:1117-1125.
[31] Hynynen K, Jolesz FA (1998). Demonstration of potential noninvasive ultrasound brain therapy through an intact skull. Ultrasound Med Biol, 24:275-283.
[32] White PJ, Clement GT, Hynynen K (2006). Longitudinal and shear mode ultrasound propagation in human skull bone. Ultrasound Med Biol, 32:1085-1096.
[33] Lee W, Croce P, Margolin RW, Cammalleri A, Yoon K, Yoo SS (2018). Transcranial focused ultrasound stimulation of motor cortical areas in freely-moving awake rats. BMC Neurosci, 19:57.
[34] Li H, Sun J, Zhang D, Omire-Mayor D, Lewin PA, Tong S (2017). Low-intensity (400 mW/cm(2), 500 kHz) pulsed transcranial ultrasound preconditioning may mitigate focal cerebral ischemia in rats. Brain Stimul, 10:695-702.
[35] Wang J, Xie L, Yang C, Ren C, Zhou K, Wang B, et al. (2015). Activated regulatory T cell regulates neural stem cell proliferation in the subventricular zone of normal and ischemic mouse brain through interleukin 10. Front Cell Neurosci, 9:361.
[36] Wang J, Lin X, Mu Z, Shen F, Zhang L, Xie Q, et al. (2019). Rapamycin Increases Collateral Circulation in Rodent Brain after Focal Ischemia as detected by Multiple Modality Dynamic Imaging. Theranostics, 9:4923-4934.
[37] Qiu W, Zhou J, Chen Y, Su M, Li G, Zhao H, et al. (2017). A Portable Ultrasound System for Non-Invasive Ultrasonic Neuro-Stimulation. IEEE Trans Neural Syst Rehabil Eng, 25:2509-2515.
[38] Tang Y, Wang J, Lin X, Wang L, Shao B, Jin K, et al. (2014). Neural stem cell protects aged rat brain from ischemia-reperfusion injury through neurogenesis and angiogenesis. J Cereb Blood Flow Metab, 34:1138-1147.
[39] He Y, Gao Y, Zhang Q, Zhou G, Cao F, Yao S (2020). IL-4 Switches Microglia/macrophage M1/M2 Polarization and Alleviates Neurological Damage by Modulating the JAK1/STAT6 Pathway Following ICH. Neuroscience, 437:161-171.
[40] Song Y, Li Z, He T, Qu M, Jiang L, Li W, et al. (2019). M2 microglia-derived exosomes protect the mouse brain from ischemia-reperfusion injury via exosomal miR-124. Theranostics, 9:2910-2923.
[41] Askari VR, Shafiee-Nick R (2019). The protective effects of beta-caryophyllene on LPS-induced primary microglia M1/M2 imbalance: A mechanistic evaluation. Life Sci, 219:40-73.
[42] Lopes RL, Borges TJ, Zanin RF, Bonorino C (2016). IL-10 is required for polarization of macrophages to M2-like phenotype by mycobacterial DnaK (heat shock protein 70). Cytokine, 85:123-129.
[43] Mansur CG, Fregni F, Boggio PS, Riberto M, Gallucci-Neto J, Santos CM, et al. (2005). A sham stimulation-controlled trial of rTMS of the unaffected hemisphere in stroke patients. Neurology, 64:1802-1804.
[44] Takeuchi N, Chuma T, Matsuo Y, Watanabe I, Ikoma K (2005). Repetitive transcranial magnetic stimulation of contralesional primary motor cortex improves hand function after stroke. Stroke, 36:2681-2686.
[45] Cullen CL, Young KM (2016). How Does Transcranial Magnetic Stimulation Influence Glial Cells in the Central Nervous System? Front Neural Circuits, 10:26.
[46] Wagner T, Valero-Cabre A, Pascual-Leone A (2007). Noninvasive human brain stimulation. Annu Rev Biomed Eng, 9:527-565.
[47] Hua Y, Jia L, Xing Y, Hui P, Meng X, Yu D, et al. (2019). Distribution Pattern of Atherosclerotic Stenosis in Chinese Patients with Stroke: A Multicenter Registry Study. Aging Dis, 10:62-70.
[48] Chang Y, Yang J, Hong H, Ma H, Cui X, Chen L (2018). The Value of Contrast-Enhanced Ultrasonography Combined with Real-Time Strain Elastography in the Early Diagnosis of Prostate Cancer. Aging Dis, 9:480-488.
[49] Pikhovych A, Stolberg NP, Jessica Flitsch L, Walter HL, Graf R, Fink GR, et al. (2016). Transcranial Direct Current Stimulation Modulates Neurogenesis and Microglia Activation in the Mouse Brain. Stem Cells Int, 2016:2715196.
[50] Braun R, Klein R, Walter HL, Ohren M, Freudenmacher L, Getachew K, et al. (2016). Transcranial direct current stimulation accelerates recovery of function, induces neurogenesis and recruits oligodendrocyte precursors in a rat model of stroke. Exp Neurol, 279:127-136.
[51] Gellner AK, Reis J, Fritsch B (2016). Glia: A Neglected Player in Non-invasive Direct Current Brain Stimulation. Front Cell Neurosci, 10:188.
[52] Perez-Asensio FJ, Perpina U, Planas AM, Pozas E (2013). Interleukin-10 regulates progenitor differentiation and modulates neurogenesis in adult brain. J Cell Sci, 126:4208-4219.
[53] Kiyota T, Ingraham KL, Swan RJ, Jacobsen MT, Andrews SJ, Ikezu T (2012). AAV serotype 2/1-mediated gene delivery of anti-inflammatory interleukin-10 enhances neurogenesis and cognitive function in APP+PS1 mice. Gene Ther, 19:724-733.
[54] Yang J, Jiang Z, Fitzgerald DC, Ma C, Yu S, Li H, et al. (2009). Adult neural stem cells expressing IL-10 confer potent immunomodulation and remyelination in experimental autoimmune encephalitis. J Clin Invest, 119:3678-3691.
[1] Xingzhi Guo,Yanying Liu,David Morgan,Li-Ru Zhao. Reparative Effects of Stem Cell Factor and Granulocyte Colony-Stimulating Factor in Aged APP/PS1 Mice[J]. Aging and disease, 2020, 11(6): 1423-1443.
[2] Yuanjian Fang,Reng Ren,Hui Shi,Lei Huang,Cameron Lenahan,Qin Lu,Lihui Tang,Yi Huang,Jiping Tang,Jianmin Zhang,John H Zhang. Pituitary Adenylate Cyclase-Activating Polypeptide: A Promising Neuroprotective Peptide in Stroke[J]. Aging and disease, 2020, 11(6): 1496-1512.
[3] Xiaoheng Li, Yajin Liao, Yuan Dong, Shuoshuo Li, Fengchao Wang, Rong Wu, Zengqiang Yuan, Jinbo Cheng. Mib2 Deficiency Inhibits Microglial Activation and Alleviates Ischemia-Induced Brain Injury[J]. Aging and disease, 2020, 11(3): 523-535.
[4] Ming Dong, Ziyi Yang, Hongcheng Fang, Jiaqing Xiang, Cong Xu, Yanqing Zhou, Qianying Wu, Jie Liu. Aging Attenuates Cardiac Contractility and Affects Therapeutic Consequences for Myocardial Infarction[J]. Aging and disease, 2020, 11(2): 365-376.
[5] Le Wang, Yang Liu, Shuxin Yan, Tianshu Du, Xia Fu, Xiaoli Gong, Xinyu Zhou, Ting Zhang, Xiaomin Wang. Disease Progression-Dependent Expression of CD200R1 and CX3CR1 in Mouse Models of Parkinson’s Disease[J]. Aging and disease, 2020, 11(2): 254-268.
[6] Wanqing Sun, Yishi Wang, Yang Zheng, Nanhu Quan. The Emerging Role of Sestrin2 in Cell Metabolism, and Cardiovascular and Age-Related Diseases[J]. Aging and disease, 2020, 11(1): 154-163.
[7] Shawn Zheng Kai Tan, Man-Lung Fung, Junhao Koh, Ying-Shing Chan, Lee Wei Lim. The Paradoxical Effect of Deep Brain Stimulation on Memory[J]. Aging and disease, 2020, 11(1): 179-190.
[8] Jinfan Tian, Sharif Popal Mohammad, Yingke Zhao, Yanfei Liu, Keji Chen, Yue Liu . Interplay between Exosomes and Autophagy in Cardiovascular Diseases: Novel Promising Target for Diagnostic and Therapeutic Application[J]. Aging and disease, 2019, 10(6): 1302-1310.
[9] Tatiana Rafaela Lemos Lima, Vívian Pinto Almeida, Arthur Sá Ferreira, Fernando Silva Guimarães, Agnaldo José Lopes. Handgrip Strength and Pulmonary Disease in the Elderly: What is the Link?[J]. Aging and disease, 2019, 10(5): 1109-1129.
[10] Kyoungjoo Cho. Emerging Roles of Complement Protein C1q in Neurodegeneration[J]. Aging and disease, 2019, 10(3): 652-663.
[11] Jianji Xu,Yunjin Zang,Dongjie Liu,Tongwang Yang,Jieling Wang,Yanjun Wang,Xiaoni Liu,Dexi Chen. DRAM is Involved in Hypoxia/Ischemia-Induced Autophagic Apoptosis in Hepatocytes[J]. Aging and disease, 2019, 10(1): 82-93.
[12] Dong Liu,Liqun Xu,Xiaoyan Zhang,Changhong Shi,Shubin Qiao,Zhiqiang Ma,Jiansong Yuan. Snapshot: Implications for mTOR in Aging-related Ischemia/Reperfusion Injury[J]. Aging and disease, 2019, 10(1): 116-133.
[13] Yang Hua,Lingyun Jia,Yingqi Xing,Pinjing Hui,Xuan Meng,Delin Yu,Xiaofang Pan,Yalan Fang,Binbin Song,Chunxia Wu,Chunmei Zhang,Xiufang Sui,Youhe Jin,Jingfen Zhang,Jianwei Li,Ling Wang,Yuming Mu,Jingxin Zhong,Yuhong Zhu,Heng Zhang,Xiaoyu Cai. Distribution Pattern of Atherosclerotic Stenosis in Chinese Patients with Stroke: A Multicenter Registry Study[J]. Aging and disease, 2019, 10(1): 62-70.
[14] Wanying Duan, Yuehua Pu, Haiyan Liu, Jing Jing, Yuesong Pan, Xinying Zou, Yilong Wang, Xingquan Zhao, Chunxue Wang, Yongjun Wang, Ka Sing Lawrence Wong, Ling Wei, Liping Liu, . Association between Leukoaraiosis and Symptomatic Intracranial Large Artery Stenoses and Occlusions: the Chinese Intracranial Atherosclerosis (CICAS) Study[J]. Aging and disease, 2018, 9(6): 1074-1083.
[15] Zhang Jun, Liu Kaiyin, Elmadhoun Omar, Ji Xunming, Duan Yunxia, Shi Jingfei, He Xiaoduo, Liu Xiangrong, Wu Di, Che Ruiwen, Geng Xiaokun, Ding Yuchuan. Synergistically Induced Hypothermia and Enhanced Neuroprotection by Pharmacological and Physical Approaches in Stroke[J]. Aging and disease, 2018, 9(4): 578-589.
Viewed
Full text


Abstract

Cited

  Shared   
Copyright © 2014 Aging and Disease, All Rights Reserved.
Address: Aging and Disease Editorial Office 3400 Camp Bowie Boulevard Fort Worth, TX76106 USA
Fax: (817) 735-0408 E-mail: editorial@aginganddisease.org
Powered by Beijing Magtech Co. Ltd