Name: diagnosis of hirschsprung's disease by immunostaining rectal suction biopsies for calretinin, s100 protein and protein gene product 9.5
Uploaded: 2019-04-26T12:30:00
Description: Watch this Scientific Journal Video about Diagnosis of Hirschsprung's Disease by Immunostaining Rectal Suction Biopsies for Calretinin, S100 Protein and Protein Gene Product 9.5 at JoVE.com

作者感谢唐伟兵在提供拍摄实验室方面的巨大帮助。本文得到了公共福利研究的支持, 并从中国国家卫生和计划生育组织获得了特别资金 (赠款号 20140: 2007)。

该协议得到了华中科技大学联合医院研究伦理委员会的批准。
1. 直肠穿刺活检
<ol><li>在获得监护人的知情同意后, 由训练有素的儿科外科医生和助理使用 Rbi2 抽吸直肠活检系统进行直肠穿刺活检。<ol>
<li>对有以下症状的患者实施 RSB: 婴儿无法通过胎粪, 或儿童长期腹胀, 有或没有便秘;长期便秘的儿童, 需要石蜡油完成排便;患有肠造口术的儿童肠梗阻或肠穿孔, 原因不明。 ?...

<ol>
	<li>Tam, P. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hirschsprung's+disease:+A+bridge+for+science+and+surgery.">Hirschsprung's disease: A bridge for science and surgery.</a> Journal of Pediatric Surgery. 51 (1), 18-22 (2016).</li><li>Heanue, T. A., Pachnis, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Enteric+nervous+system+development+and+Hirschsprung's+disease:+advances+in+genetic+and+stem+cell+studies.">Enteric nervous system development and Hirschsprung's disease: advances in genetic and stem cell studies.</a> Nature Reviews Neuroscience. 8 (6), 466-479 (2007).</li><li>Setiadi, J. A., Dwihantoro, A., Iskandar, K., Heriyanto, D. S., Gunadi, <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+utility+of+the+hematoxylin+and+eosin+staining+in+patients+with+suspected+Hirschsprung+disease.">The utility of the hematoxylin and eosin staining in patients with suspected Hirschsprung disease.</a> BMC Surgery. 17 (1), 71 (2017).</li><li>Agrawal, R. K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acetylcholinesterase+histochemistry+(AChE)+-+A+helpful+technique+in+the+diagnosis+and+in+aiding+the+operative+procedures+of+Hirschsprung+disease.">Acetylcholinesterase histochemistry (AChE) - A helpful technique in the diagnosis and in aiding the operative procedures of Hirschsprung disease.</a> Diagnostic Pathology. 10 (1), 208 (2015).</li><li>Kacar, A., Arikok, A. T., Azili, M. N., Ekberli Agirbas, G., Tiryaki, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Calretinin+immunohistochemistry+in+Hirschsprung's+disease:+An+adjunct+to+formalin-based+diagnosis.">Calretinin immunohistochemistry in Hirschsprung's disease: An adjunct to formalin-based diagnosis.</a> The Turkish Journal of Gastroenterology. 23 (3), 226-233 (2012).</li><li>Bachmann, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immunohistochemical+panel+for+the+diagnosis+of+Hirschsprung&#39;s+disease+using+antibodies+to+MAP2,+calretinin,+GLUT1+and+S100.">Immunohistochemical panel for the diagnosis of Hirschsprung&#39;s disease using antibodies to MAP2, calretinin, GLUT1 and S100.</a> Histopathology. 66 (6), 824-835 (2015).</li><li>Jiang, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=S100+and+protein+gene+product+9.5+immunostaining+of+rectal+suction+biopsies+in+the+diagnosis+of+Hirschsprung'+disease.">S100 and protein gene product 9.5 immunostaining of rectal suction biopsies in the diagnosis of Hirschsprung' disease.</a> American Journal of Translational Research. 8 (7), 3159 (2016).</li><li>Takawira, C., D'Agostini, S., Shenouda, S., Persad, R., Sergi, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laboratory+procedures+update+on+Hirschsprung+disease.">Laboratory procedures update on Hirschsprung disease.</a> Journal of Pediatric Gastroenterology &amp; Nutrition. 60 (5), 598 (2015).</li><li>Meier-Ruge, W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acetylcholinesterase+activity+in+suction+biopsies+of+the+rectum+in+the+diagnosis+of+Hirschsprung's+disease.">Acetylcholinesterase activity in suction biopsies of the rectum in the diagnosis of Hirschsprung's disease.</a> Journal of Pediatric Surgery. 7 (1), 11-17 (1972).</li><li>Barshack, I., Fridman, E., Goldberg, I., Chowers, Y., Kopolovic, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+loss+of+calretinin+expression+indicates+aganglionosis+in+Hirschsprung's+disease.">The loss of calretinin expression indicates aganglionosis in Hirschsprung's disease.</a> Journal of Clinical Pathology. 57 (7), 712-716 (2004).</li><li>Kapur, R. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Can+We+Stop+Looking?+Immunohistochemistry+and+the+Diagnosis+of+Hirschsprung+Disease.">Can We Stop Looking? Immunohistochemistry and the Diagnosis of Hirschsprung Disease.</a> American Journal of Clinical Pathology. 126 (1), 9-12 (2006).</li><li>Guinardsamuel, V., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Calretinin+immunohistochemistry:+a+simple+and+efficient+tool+to+diagnose+Hirschsprung+disease.">Calretinin immunohistochemistry: a simple and efficient tool to diagnose Hirschsprung disease.</a> Modern Pathology. 22 (10), 1379-1384 (2009).</li><li>Robey, S. S., Kuhajda, F. P., Yardley, J. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immunoperoxidase+stains+of+ganglion+cells+and+abnormal+mucosal+nerve+proliferations+in+Hirschsprung's+disease.">Immunoperoxidase stains of ganglion cells and abnormal mucosal nerve proliferations in Hirschsprung's disease.</a> Human Pathology. 19 (4), 432-437 (1988).</li><li>Monforte-Mu&#241;oz, H., Gonzalez-Gomez, I., Rowland, J. M., Landing, B. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Increased+submucosal+nerve+trunk+caliber+in+aganglionosis:+a+&amp;#34;positive&amp;#34;+and+objective+finding+in+suction+biopsies+and+segmental+resections+in+Hirschsprung's+disease.">Increased submucosal nerve trunk caliber in aganglionosis: a &amp;#34;positive&amp;#34; and objective finding in suction biopsies and segmental resections in Hirschsprung's disease.</a> Archives of Pathology &amp; Laboratory Medicine. 122 (8), 721-725 (1998).</li><li>Bachmann, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immunohistochemical+panel+for+the+diagnosis+of+Hirschsprung&#39;s+disease+using+antibodies+to+MAP2,+calretinin,+GLUT1+and+S100.">Immunohistochemical panel for the diagnosis of Hirschsprung&#39;s disease using antibodies to MAP2, calretinin, GLUT1 and S100.</a> Histopathology. 66 (6), 824-835 (2015).</li><li>Sams, V. R., Bobrow, L. G., Happerfield, L., Keeling, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+PGP9.5+in+the+diagnosis+of+Hirschsprung's+disease.">Evaluation of PGP9.5 in the diagnosis of Hirschsprung's disease.</a> Journal of Pathology. 168 (1), 55 (1992).</li><li>Huang, Y., Anupama, B., Zheng, S., Xiao, X., Chen, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+expression+of+enteric+nerve+markers+and+nerve+innervation+in+total+colonic+aganglionosis.">The expression of enteric nerve markers and nerve innervation in total colonic aganglionosis.</a> International Journal of Surgical Pathology. 19 (3), 303 (2011).</li></ol>

在这里, 我们描述了一个程序, 使用三种不同的免疫组织化学抗体染色 RBS 部分诊断亨廷顿舞蹈症。我们的诊断方案的敏感性为 99.49% (95% CI, 0.88-0.99), 特异性为 100% (95% CI, 0.88-1.00)。
该协议最关键的步骤是 RSB 和抗原抗体反应。活检的大小决定了染色的准确性。小活检不会提供足够的组织来做出准确的诊断, 而大的活检会导致更高的出血风险。根据经验, 直径为2毫米的活检是最佳?...

先天性巨结肠病 (HD) 是一种常见的先天性肠道疾病, 其特点是肠远端1的不同部分缺乏神经节细胞.胚胎神经细胞的入侵完成后, 就形成了人类肠道神经系统。如果有一个过程的干扰, 并且入侵未能完成, 新生儿的远端肠成为琼脂2。这种潜在的致命疾病被称为先天性巨结肠病。增殖、运动和肠道生长是成功殖民的三个主要组成部分。
传统的半粘膜下活检的血起毒素和 eosin (H & E) 染色不能像手术中获得的全厚度组织的 h & E 染色那样令人满意。此外, 直肠吸力组织的乙酰胆碱酯酶 (ache) 染色由于其灵敏度不足 (91%) 和冷冻切片 3, 4 的复杂处理, 在理论上具有挑战性。神经节细胞和神经纤维的其他几个免疫组织化学标志物, 可以染色的福尔马林固定和石蜡嵌入标本正在逐渐成为主流的亨廷顿舞蹈症诊断。卡莱丁素是一种维生素 d 依赖性钙结合蛋白, 不表达在肌肠内和粘膜下神经丛的 hd 影响段 5.S100 蛋白在神经冠产生的细胞中表达, 如神经纤维和胶质细胞, 这些细胞通常在 hd 影响的段的粘膜下组织中表现出神经肥大 6.蛋白质基因产品 9.5 (PGP9.5) 可靠地染色神经纤维和神经节细胞;PGP9.5 染色是对卡氏素染色的补充, 特别是在孤立的性腺功能减退症的情况下。双染色 S100 和 PGP9.5 可以降低假阴性率, 提高灵敏度。作为一个前提, 目前的研究旨在确保充分的特异性和高灵敏度的这种新的诊断方法。我们的新协议使用了所有三个标记的鉴别共济术肠和肥厚神经纤维。我们的实验室对318个儿童进行了前瞻性研究, 此前没有详细的协议7发表。本文讨论了详细的协议和注意事项。任何新生儿从出生起就患有严重的排便问题, 或患有慢性便秘的儿童, 不包括其他常见疾病, 都是直肠抽吸活检 (RSB) 的潜在候选人。我们的新方案不仅适用于动态实验室染色, 还适用于全厚度活检或手术标本的染色, 以做出最终诊断。

<table>
	<tbody>
		<tr>
			<td>calretinin antibody</td>
			<td>MXB Biotechnologies</td>
			<td>MAB-0716 170416405c</td>
			<td>antibody: primary antibody</td>
		</tr>
		<tr>
			<td>S-100 antibody</td>
			<td>MXB Biotechnologies</td>
			<td>Kit-0007</td>
			<td>antibody: primary antibody</td>
		</tr>
		<tr>
			<td>PGP9.5 antibody</td>
			<td>Shanghai long island antibody Co. Ltd</td>
			<td>R-0457-03</td>
			<td>antibody: primary antibody</td>
		</tr>
		<tr>
			<td>enhancer reagent</td>
			<td>MXB Biotechnologies</td>
			<td>KIT-9902-A 170416405a</td>
			<td>antibody: secondary antibody A</td>
		</tr>
		<tr>
			<td>Goat anti-Rabbit/Mouse IgG Secondary Antibody</td>
			<td>MXB Biotechnologies</td>
			<td>KIT-9902-B</td>
			<td>antibody: secondary antibody B</td>
		</tr>
		<tr>
			<td>DAB staining kit (containing reagent A B and C)</td>
			<td>MXB Biotechnologies</td>
			<td>DAB-0031</td>
			<td>staining kit</td>
		</tr>
		<tr>
			<td>Heat incubator</td>
			<td>Shanghai yiheng instrument Co. Ltd</td>
			<td>DHP-9082</td>
			<td>instrument</td>
		</tr>
		<tr>
			<td>Rbi2 suction rectal biopsy system</td>
			<td>Aus Systems Pty Ltd, South Australia, Australia</td>
			<td>CP1200 HP1000 SS1000</td>
			<td>instrument</td>
		</tr>
		<tr>
			<td>microtome</td>
			<td>Leica</td>
			<td>leica RM2016</td>
			<td>instrument</td>
		</tr>
		<tr>
			<td>citric acid sodium citrate buffer(100X)</td>
			<td>MXB Biotechnologies</td>
			<td>MVS-0101</td>
			<td>antigen retrieval buffer</td>
		</tr>
		<tr>
			<td>pathological tissue dehydrator</td>
			<td>wuhan junjie electronic Co. Ltd</td>
			<td>JT-12F</td>
			<td>instrument</td>
		</tr>
	</tbody>
</table>

diagnosis of hirschsprung's disease by immunostaining rectal suction biopsies for calretinin, s100 protein and protein gene product 9.5

先天性巨结肠病 (HD) 是一种先天性肠道疾病, 临床表现为婴儿无法通过胎粪或儿童长期便秘。直肠抽吸活检 (RSB) 测定神经节细胞的缺失和神经肥大是目前诊断亨廷顿舞蹈症最准确的方法。传统的血红素-eosin 染色缺乏敏感性和特异性。乙酰胆碱酯酶染色由于其复杂的过程, 不能被广泛使用。我们在 rsb 上进行的用于卡利丁、S100 蛋白和蛋白质基因产物 9.5 (PGP9.5) 的免疫染色新协议, 具有 96.49 (95% 置信区间、0.88-0.99) 和 100% (95% 置信度) 的高灵敏度和特异性。间隔, 分别为 0.97-1.00)。受 hd 影响的片段通常表现为缺乏卡莱丁、S100 蛋白和 PGP9.5 的表达, 而这些都是黏膜下组织中神经肥大的标志物。此协议描述了这种新诊断方法的详细操作过程。

共有318名患者参加了我们的研究。所有患者均接受 RSB 治疗, 并对组织进行了卡瑞丁、S100 和 PGP9.5 的染色。基于我们的新协议的诊断是 HD 97 例, 非 hd 213 例, 疑似 HD 8 例。在132例手术患者中, 99 例经手术后对全厚度标本进行免疫染色, 诊断为亨廷顿舞蹈症。S100 和 PGP9.5 显示, 在99例患者中, 92% 和93% 的患者分别有神经干肥大。186名患者中, 其余83例接受了全厚度活检, 并显示了神...

Watch this Scientific Journal Video about Diagnosis of Hirschsprung's Disease by Immunostaining Rectal Suction Biopsies for Calretinin, S100 Protein and Protein Gene Product 9.5 at JoVE.com

Diagnosis of Hirschsprung's Disease by Immunostaining Rectal Suction Biopsies for Calretinin, S100 Protein and Protein Gene Product 9.5

该方案描述了对卡莱丁、S100 蛋白和蛋白质基因产物9.5 进行免疫染色直肠抽吸活检的过程。这种新型辅助诊断方法对先天性巨结肠病具有较好的敏感性和特异性。

卡瑞津、S100 蛋白和蛋白质基因产物9.5 的免疫修饰直肠吸吮活检诊断

Hirschsprung&#39;s disease (HD) is a congenital intestinal disease that is clinically manifested as an inability to pass meconium in infants or as ...

这种方法可以帮助回答赫希斯普龙病的诊断问题。如果遵循协议，该技术的方法和功能会表现出高灵敏度和同时处理的特异性率。演示程序将是米景方，一个技术人员从我的实验室。首先将直肠吸活检固定在4%半甲醛中组织体积的五倍，在室温下6小时。将固定组织脱水 11 小时，并将加工过的直肠吸活检组织嵌入石蜡块中。修剪微原子上的块，直到组织的完整截面被暴露。然后使用微原子获得0.4微米的嵌入组织部分。将45至50摄氏度的水，让部分铺成平面图。将扁平部分转移到玻璃滑梯上，在 60 摄氏度的烤箱中烘烤三到五个小时。将脱甲脱素的幻灯片放入两个 15 分钟的二甲苯变化中，然后连续进行 5 分钟乙醇浸入。然后用反渗透纯净水冲洗滑梯五分钟。对于抗原检索，将幻灯片放入可检索溶液的科普林染色罐中，然后将罐子放入沸水的预热浴中。20 分钟后，让罐子冷却到室温约 10 分钟，然后用 PBS 轻轻冲洗幻灯片。然后使用疏水标记包围每个组织部分，并放入湿盒中。要阻止任何过氧化物，请在每个组织上加入两到三滴 3% 的过氧化氢溶液，并将幻灯片在 37 摄氏度下放置 20 分钟。在孵育结束时，用三个五分钟的PBS洗涤冲洗幻灯片，然后用5%牛血清白蛋白在37摄氏度下阻断20分钟。接下来，丢弃过量的牛血清白蛋白，并在4摄氏度下用50微升的原抗体标记细胞。第二天早上，用PBS三分钟洗涤液冲洗幻灯片，并在每张幻灯片中加入50微升聚合物增强剂。在37摄氏度下20分钟后，用PBS清洗幻灯片三次，并在每个组织部分添加50微升的二次抗体B。在 37 摄氏度下 30 分钟后，用 PBS 中三个 5 分钟的洗涤液冲洗部分，并在每张幻灯片上添加一滴新鲜准备的 DAB 工作解决方案。当通过光显微镜检测到适当水平的棕色染色时，在 PBS 中冲洗幻灯片 5 分钟，然后用一滴赤氧林对核进行反染色一分钟。去除水滴下多余的污渍约 30 至 40 秒，然后用新鲜 PBS 洗涤 25 到 30 秒。在1%盐酸乙醇中区分组织10秒，然后洗1分钟PBS。以水化的相反顺序浸入五分钟使幻灯片脱水，使幻灯片暴露在两个五分钟二甲苯的变化中。然后使用超清洁安装安装盖滑，让幻灯片在光显微镜上可视化之前干燥。在这项对318例直肠部分活检患者的代表性研究中，99名患者最初被诊断为直肠部分活检。在正常组织活检中观察到许多表达卡雷丁素的结节细胞。然而，在赫希斯普龙病段的组织的任何区域中未检测到结节细胞。S-100 和 PGP9.5 免疫染色显示，在患病组织的亚木科萨中，神经躯干会肥大，在正常内肠中观察到的一些小神经树枝只有颗粒染色。在尝试此程序时，重要的是要记住注意细节，如直肠吸活检的大小和部分的厚度。在碎片整理后，这项技术将为儿科外科领域的研究铺平道路，以支持赫希斯普龙病的诊断。

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Spinal Cord Electrophysiology II: Extracellular Suction Electrode Fabrication

脊髓电二：外吸电极的制作

Development of neural circuitries and locomotion can be studied using neonatal rodent spinal cord central pattern generator (CPG) behavior. We demonstrate ...

Detection of Microregional Hypoxia in Mouse Cerebral Cortex by Two-photon Imaging of Endogenous NADH Fluorescence

检测内源NADH的荧光双光子成像的小鼠大脑皮层Microregional缺氧

The brain's ability to function at high levels of metabolic demand depends on continuous oxygen supply through blood flow and tissue oxygen diffusion. ...

Sequential Photo-bleaching to Delineate Single Schwann Cells at the Neuromuscular Junction

连续光漂白划定单雪旺细胞在神经肌肉接头处

Sequential photo-bleaching provides a non-invasive way to label individual SCs at the NMJ. The NMJ is the largest synapse of the mammalian nervous system ...

Detection of Protein Palmitoylation in Cultured Hippocampal Neurons by Immunoprecipitation and Acyl-Biotin Exchange (ABE)

Detection of Protein Palmitoylation in Cultured Hippocampal Neurons by Immunoprecipitation and Acyl-Biotin Exchange ABE

检测蛋白质棕榈酰化培养海马神经元通过免疫沉淀和酰基-生物素交易所（ABE）

Palmitoylation is a post-translational lipid  modification involving the attachment of a 16-carbon saturated fatty acid,  palmitate, to cysteine residues ...

Detection of In Situ Protein-protein Complexes at the Drosophila Larval Neuromuscular Junction Using Proximity Ligation Assay

Detection of In Situ Protein-protein Complexes at the Drosophila Larval Neuromuscular Junction Using Proximity Ligation Assay

检测<em&gt;原位</em&gt;蛋白质 - 蛋白质复合物的<em&gt;果蝇</em&gt;幼虫神经肌肉接头使用接近结扎分析

Discs large (Dlg) is a conserved member of the membrane-associated guanylate kinase family, and serves as a major scaffolding protein at the larval ...

The Neuromuscular Junction: Measuring Synapse Size, Fragmentation and Changes in Synaptic Protein Density Using Confocal Fluorescence Microscopy

神经肌肉接头处：测量突触大小，碎片和改变突触蛋白密度利用共聚焦荧光显微镜

The neuromuscular junction (NMJ) is the large, cholinergic relay synapse through which mammalian motor neurons control voluntary muscle contraction. ...

Immunostaining of Biocytin-filled and Processed Sections for Neurochemical Markers

对于神经化学标记用biocytin填充和加工部分的免疫染色

Electrophysiological recordings of cells using the patch clamp technique have allowed for the identification of different neuronal types based on firing ...

Bioluminescence Imaging of Neuroinflammation in Transgenic Mice After Peripheral Inoculation of Alpha-Synuclein Fibrils

在转基因小鼠神经炎症的生物发光成像的α-共核蛋白纤丝的外围接种后

To study the prion-like behavior of misfolded alpha-synuclein, mouse models are needed that allow fast and simple transmission of alpha-synuclein ...

Live Images of GLUT4 Protein Trafficking in Mouse Primary Hypothalamic Neurons Using Deconvolution Microscopy

用反褶积显微镜观察小鼠原发性下丘脑神经元 GLUT4 蛋白的动态图像

Type 2 diabetes mellitus (T2DM) is a global health crisis which is characterized by insulin signaling impairment and chronic inflammation in peripheral ...