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A rodent model of acute myocardial infarction(MI) was first developed in the rat[7].More recently a murine equivalent has been described[8],providing the means to exploit the increasing availability of many useful transgenic and knockout mouse strains. Complete occlusion of the left anterior descending(LAD)coronary artery induces an acute MI. The resulting ischaemia in the left ventricular wall has been visualized with Evans blue/TTC perfusion assays[9-12],which allows quantification of the infarct area. Further work by Guo et al.[13] has demonstrated ischaemic preconditioning after short-term LAD occlusion in mouse, thus validating the physiologic relevance of this infarct size. Aminoacyl-tRNA synthetases catalyze the first step of protein synthesis that consists of the aminoacylation of tRNAs. But they have a broad repertoire of functions beyond protein synthesis, including transcriptional and translational regulation as well as cell signaling [14]. Recently, it has been demonstrated that two of the tRNA synthetases, human tyrosyl-tRNA synthetase (TyrRS) and human tryptophanyl-tRNA synthetase (TrpRS), have novel cytokine functions[15]. This demonstration established a link between protein synthesis and signal transduction. At the same time,mammalian TyrRS and TrpRS have also been shown to regulate angiogenesis [16-21]. Under apoptotic conditions in culture, full lengthTyrRS is secreted, and two distinct cytokines can then be generated by an extracellular protease such as leukocyte elastase[15].The NH2-terminal catalytic fragment, mini-TyrRS, binds strongly to the CXC-chemokine receptor CXCR1 and, like IL-8, functions as a chemoattractant for polymorphonuclear leukocytes (PMNs), to promote angiogenesis[15], whereas the full-length enzyme lacks cytokine activity.The catalytic core domain of TrpRS is a close homologue of the catalytic domain of TyrRS [22-24]. In normal cells, human TrpRS exists as two forms. The major form is the full-length protein, and the other is a truncated TrpRS (mini-TrpRS) in which most of the extra NH2-terminal domain is deleted because of alternative splicing of the pre-mRNA[25-26], with Met-48 being deduced as the NH 2 -terminal residue of mini- TrpRS [25]. PMN elastase digestion of recombinant full-length TrpRS produced two fragments designated T 1 -TrpRS and T 2 -TrpRS, respectively.These fragments were similar in size to mini-TrpRS. In addition, mini-TrpRS and T2-TrpRS blocked vascular endothelial growth factor-stimulated angiogenesis in both chick cell adhesion molecule and mouse matrigel assays in vivo [18-19]. The full-length enzyme lacks cytokine activities. The construction and relationship of TyrRS, TrpRS, and their variants is shown in Fig. 1. Fig 1. Schematic representation of human TyrRS, TrpRS and their variant constructs used in this study. Shaded regions of full-length TyrRS and TrpRS represent COOH- and NH 2 -terminal appended domains, respectively. Numbers on the left and right correspond to the NH 2 - and COOH-terminal residues relative to the human full-length enzymes, respectively. Despite these intriguing in vitro actions, no studies have examined the mini-TyrRS and mini- TrpRS in physiological or pathological settings in vivo, especially in myocardial ischemia condition. Therefore, the purpose of this study was to determine whether exogenous mini-TyrRS augments angiogenesis while mini-TrpRS inhibited, and to preliminary comprehend their angiogenesis mechanism. All of this may be helpful for healing ischemia diseases, such as CAHD. 2. Methods 38
2.1 Materials Sprague-Dauley(SD) male rats,(250-300g, 2-3 month old) were provided from experimental animal center of Sichuan university.(Sichuan, China). Human recombinant mini-TyrRS and mini- TrpRS were from aTyr Pharma (La Jolla, CA).Ⅷ factor related antigen and antibody was from Boster(Wuhan, china). Immunohistochemisty staining kit was purchased from Zhongshan Goldenbridge (Beijing, China). Revert Aid First Strand cDNA Synthesis Kit was purchased from MBI Company (Lithuania). 2.2.Ethics All animal procedures were conducted with prior institutional ethical approval under the requirements of the Chinese Prevention of Cruelty to Animals Act and the Code of Practice for the Care and Use of Animals for Scientific Purposes. Prior clearance was obtained from the Animal Experimentation Ethics Committees of West china Medical Centre and Institutes of Animal Science. The animals of this study were inspected by members of the West china Medical Centre Animal Ethics Committee. 2.3.Left anterior descending (LAD) artery ligation Animals were anaesthetized with 100g/L chloral hydrate (0.2ml per 100g body weight injected intraperitoneally).The analgesic buprenorphine was given preoperatively(10-20 µg/kg body weight).Occlusion of the left anterior descending coronary artery(LAD) was performed as previously reported[11],under sterilec conditions, with minor alterations. Briefly,the anaesthetized animal was incubated endotracheally in a supine position, and ventilated with a Harvard Mouse Mini- Vent(Harvard apparatus, Marchhugstetten,Germany),which supplied 0.2-0.25ml room air 120 times per minute. The animal was moved onto its right side, and a left thoracotomy in the third intercostal space was provided access to the beating heart. After removing the pericardium,the LAD was visualized with a stereomicroscope (Leica MZ6,Heerbrugg,Switzerland),and occluded with 8/0 prolene suture. The suture position of the LAD coronary artery was 0.3mm distal to the atrioventricular junction. Occlusion was confirmed by observation of left ventricular pallor immediately post ligation and an electrocardiogram was used to observe changes such as widening of QRS and ST-T segment elevation. The chest was closed, the lungs re-inflate and the animal moved to a prone position until spontaneous breathing occurred. Animal were monitored closely for signs of infection at the surgical site; none were observed in any animals. 2.4.Experimental groups A total of 80 rats(200-250g) were divided into four different groups(n=20 per each group, and n=5 for each time point).⑴sham group: animals underwent a thoracotomy with removal of the pericardium, but no coronary artery ligation (CAL). No suture was placed in the sham animals’heart, in order to avoid unintended vessel damage or occlusion; ⑵ CAL group, but no mini-TyrRS/mini- TrpRS-siRNA injection;⑶: CAL+mini-TyrRS(20μl, twice daily, 600μg. Kg -1 .day -1 ) ⑷: CAL+mini- TrpRS(20μl, twice daily, 600μg. Kg -1 .day -1 ). Mini-TyrRS or mini-TrpRS were administrated by coronary artery polyvinyl catheter injection to rats. 2.5.Histologic expression All rats were killed after ligation 3rd, 7th, 14th, and 28th day ,the heart was removed and fixed in fresh 4% paraformaldehyde, pH 7.4.The tissue was processed and embedded in paraffin using routine histological procedures. Five micrometre transverse step sections were collected every 200µm through the entire ventricle(approximately 10-12 sections per animal),and stained with Haematoxylin and Eosin(HE).the cells were observed with an inverted phase-contrast microscope (Olympus, Japan) and photographed. The total tube area were analyzed in 3 different fields at ×400 magnification 2.6.Measurement of irreversible ischemic injury After ligation 3rd, 7th, 14th, and 28th day, 1% of Evans blue solution (5 mL),was infused into the abdominal vena cava to delineate the ischemic area at risk of the left ventricle. The heart was excised and cross-sectioned from the apex to the atrioventricular groove into four specimens of 0.8mm in thickness with the use of a stereoscope. The two middle heart slices were incubated in 2,3,5- 39
Page 2 and 3: The 3 rd VACPS Research Workshop Pr
Page 4 and 5: Organizing Committee Victorian Asso
Page 6 and 7: Preface On the occasion of the publ
Page 8 and 9: Paper as a low-cost base material f
Page 10 and 11: Acknowledgements We gratefully ackn
Page 13 and 14: Career planning and Job Interview i
Page 15 and 16: The role of mitochondria in atrial
Page 18 and 19: Track-Before-Detect Procedures for
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Page 42 and 43: Demonstration and Performance Analy
Page 45: Effect of mini-tyrosl-tRNA syntheta
Page 49 and 50: Fig. 2. The representative photomic
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Page 53 and 54: 3.4.The mRNA expression of mini-Tyr
Page 55 and 56: 9.Du XJ, Gao X, Ramsey D. Surgical
Page 57 and 58: Distribution of three forms alpha-m
Page 59 and 60: Materials and Methods Ethics All an
Page 61 and 62: Fig.1 (A) Mass Spectrometry results
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Page 65 and 66: Fig.6 (A) Mass Spectrometry results
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Page 69 and 70: Investigation of the mechanism of t
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Page 73: Paper as a low-cost base material f
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Page 84 and 85: A Multi-modal Gesture Recognition S
Page 86: Searching for Fair Joint Gains in A
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Grain refinement of pure magnesium
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Fig. 3 The optical and SEM microstr
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Conclusions (1) The large grains in
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[22-25]. Sorting of normal and Babe
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Figure 1. Г×Re[fCM] vs. frequency
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Figure 7. SEM image of damaged elec
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References: [1] Xiong Liu, Mark C.
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new tissue[10]. Most of these human
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2.8 Viability and morphology study
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Fig 3 and 4 SEM images of electrosp
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Fig 10 Tensile Stress curve of elec
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Uniform Coating of WO x on TiO 2 Na
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Union as a Social Regulator of Mark
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enterprises…There obviously is no
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Organizational Capabilities • Lea
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Table 1. Correlations, Reliabilitie
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[18]. Jap, S. D. (1999). Pie-expans
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Early Childhood Education Matters:
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decisions on selection of child car
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Noble, K. (2007). Parent choice of
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An Exploration of Country of Origin
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Supercontinuum generation for fiber
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Gas chromatographic retention indic
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Fiber Nonlinearity Precompensation
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