本研究得到了国家自然科学基金（81973569、82274207、82104533）、成都中医药大学星林学者研究推进项目（XKTD2022013）和宁夏市重点研发计划（2023BEG02012）的支持。

红景天 采集自中国四川省甘孜藏族自治州甘孜县卓达雪山（北纬31.44570°，东经99.96086°，海拔4892米）。这些植物由成都中医药大学民族医学院的张毅教授鉴定为真品。
1. 红景天的收集
<ol><li>拍摄 红景天的栖息地地图。</li>
	<li>拍摄红景天的整个植物、叶子、花萼和根茎。</li>
	<li>用铁锹清除红 景天 1 m 范围内的杂草和碎石，以确保随后的顺利开采。</li>
	<li>用锄头挖土，直到看到整个根际并收集主根。 注:用于药用部位的 红景天 的根和根茎应在秋季花茎枯萎时采集。</li>
</ol>2.特征识别
<ol><li>用肉眼观察 红景天 的外观特征:圆柱形和短主根和根茎，棕色表面，带有粉红色图案的膜黄色表皮以及橙红色或勃艮第切片。</li>
	<li>通过气味识别它:靠近鼻子时会散发出芬芳的气味。</li>
	<li>按味辨:取一小块根放入口中，先啜饮后咀嚼，略带苦味，后甜。</li>
</ol>3.药粉中淀粉颗粒的显微鉴定
<ol><li>用刷子去除 红景天 表面的土壤，将其放入40°C的烤箱中，每24小时翻动一次草药。 注意:药材的易碎性被认为是干燥水分的标准。</li>
	<li>用粉剂机将干燥的药材粉化，并用药用的3号筛子过滤药粉（见 材料表）。</li>
	<li>取一个干净的载玻片（见 材料表），用解剖针挖出粉末（见 材料表），并将其均匀地放置在载玻片的三分之一处，在2毫米以内。</li>
	<li>使用玻璃滴管（见 材料表）向粉末中加入一滴去离子水。使用镊子（见 材料表）握住盖玻片的一端（见 材料表），快速接触液面并覆盖粉末。 注意: 使用解剖针轻轻混合水和药粉，以确保样品混合均匀。载玻片、粉末和盖玻片之间不应有气泡。</li>
	<li>打开显微镜（参见 材料表）并将步骤3.4中的载玻片放在平台上以固定它。调整光源和粗焦螺旋以查看粉末。调整细小的旁焦点螺旋，直到清楚地看到组织。切换到40倍物镜并观察淀粉颗粒。 注意:对呈现为单个或多个颗粒的谷物进行淀粉，脐带点呈人字形或裂缝状。</li>
</ol>4.药粉中导管、软木细胞、纤维、木实质细胞、色素团的显微鉴定
<ol><li>取一张干净的载玻片（见 材料表），用解剖针挖出粉末（见 材料表），并将其放置在载玻片的三分之一处。</li>
	<li>使用玻璃滴管（见 材料表）向粉末中加入一滴水合氯醛（见 材料表）。用镊子取载玻片（见 材料表），在酒精灯中加热三次，每次加热1秒。 注意: 加热过程中应避免气泡。液体保持不流动，表明渗透已完成。</li>
	<li>使用玻璃滴管加入一滴甘油（见 材料表）。用镊子握住盖玻片的一端，快速接触液位。</li>
	<li>打开显微镜并将载玻片放在平台上以固定它。调整光源和粗焦螺旋以查看粉末。调整细小的旁焦点螺旋，直到清楚地看到组织。通过切换到 40 倍物镜观察导管、软木细胞、纤维、木实质细胞和颜料块。 注:软木细胞多边形或长多边形，螺旋状血管具有明显的螺旋结构，木质部薄壁组织含有草酸钙砂晶，红色或棕红色色素块。</li>
</ol>5. 红景天薄层色谱（TLC）样品的制备及其对照品
<ol><li>将称重纸放在天平上（见 材料表）并称取 3 克 红景天粉末。</li>
	<li>将粉末放入 100 mL 锥形瓶中（参见 材料表），并用大腹移液管加入 25 mL 甲醇（参见 材料表）。将锥形瓶放入超声波仪器中。将功率设置为 250 W，频率设置为 40 kHz，时间设置为 30 分钟（参见 材料表），然后打开仪器。 注:超声仪器的目的是确保 红景天 粉末完全溶解，而不影响后续薄层色谱实验的结果。</li>
	<li>取出锥形瓶，用流水冲洗外瓶至室温 （RT）。</li>
	<li>用 1 mL 注射器吸出步骤 5.3 中制备的 800 μL 液体。用0.22μm微孔滤膜过滤（参见 材料表），并在色谱样品瓶中收集400μL 红景天 中游样品溶液。</li>
	<li>称取并分别加入 2 mg 水景苷、酪醇和没食子酸（参见 材料表）到 3 个单独的 100 mL 锥形瓶中。用大肚移液管向每个锥形瓶中加入 25 mL 甲醇。</li>
	<li>将锥形瓶放入超声波仪器中，将功率设置为250 W，频率设置为40 kHz，时间设置为30分钟，然后重复步骤5.3（见 材料表）。</li>
	<li>用 1 mL 注射器吸出步骤 5.6 中制备的 800 μL 液体。用0.22μm微孔滤膜过滤（见 材料表），并在相应的色谱样品瓶中收集400μL中游水景苷，酪醇和没食子酸标准溶液。</li>
</ol>6. TLC鉴定
<ol><li>移液器三氯甲烷 （5 mL）、乙酸乙酯 （4 mL）、甲醇 （2 mL） 和甲酸 （0.5 mL）（参见 材料表）。加入双罐层析筒的一侧（见 材料表），摇匀并混合。盖住上气缸盖。</li>
	<li>将没食子酸，水景苷，酪醇标准溶液和 红景天 溶液放在样品架上的位置A1-A4。</li>
	<li>将 5 cm x 10 cm 硅胶片（参见 材料表）放在采样台上。启动自动取样机（见 物料表），打开气控阀。</li>
	<li>打开visionCATS软件（参见材料表）。单击"新建">"新建文件夹（名为红景天样本测试）">"确定"。单击"新方法"（名称 Rhodiola crenulata 样品测试）> OK > ATS 4。</li>
	<li>单击 "完成步骤定义"（Finish Step Definition）。单击 "跟踪分配 "以编辑描述（没食子酸、水景苷、酪醇和 红景天 样品）。</li>
	<li>单击 HPTLC 步骤。设置薄层参数 （5 cm x 10 cm）。选择 应用类型 （波段），设置参数（表1），单击 "确定 "按钮。</li>
	<li>开放式 灌装/冲洗质量。选中 仅填充编程音量。将样品瓶底部液位 （mm） 设置为 0.5，然后单击 OK 按钮。</li>
	<li>单击 "执行方法"。</li>
	<li>单击 "跟踪分配 "按钮，选中 "居中"，然后设置参数（表 2）。</li>
	<li>单击 "继续 "按钮进行自动采样。</li>
	<li>关闭自动取样机和空气阀。从自动取样机上取下硅胶片。</li>
	<li>在步骤6.1中将硅胶片放入双罐层析筒的另一侧，盖上上筒盖，预饱和20分钟。</li>
	<li>用镊子轻轻夹住薄层板的上端，迅速将硅胶片放入显影剂中，盖上上气缸盖。 注意: 当展开的前边缘距离薄层板上端 0.5-1.0 厘米时，取出硅胶片。</li>
	<li>有机溶剂蒸发后，在室温下将显色溶液喷洒到硅胶片表面，以获得显色效果。 注:显色溶液是含有 2% FeCl3 和 1% K3[Fe（CN）6] 的水溶液。</li>
</ol>

红景天月状物的显微表征和基于 TLC 的鉴定

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作者没有什么可透露的。

世界上有90多种红景天植物，其中60%以上的物种分布在我国，常见的有红景天、红景天、红景天和红景天17。《中国药典》（2020年）第一部分记载的红景天是一种生长在高海拔地区的传统藏药。红景天的市场需求每年都在增加，因此确保正确使用来源是确保安全使用的关键。特别是，从现场拣选到简单快速的实验室常规鉴定的标准化不容忽视。?...

草药在中国有着悠久的历史和丰富的应用经验，是神农草药经典1中的第一个系统记录。青蒿素应用于疟疾的发现，推动了草药的发展进入一个新的阶段1。利用现代科学技术揭示草药的确切机理，提高了草药的利用率和需求，为其开辟了新的国际市场2,3,4。然而，这导致了一系列负面影响。非专业人士对草药的特性认识模糊，这使得草药的使用面临巨大的安全风险5。
红景天是红景天属植物之一，主要分布于西藏、云南西北部和四川西部（图1）6,7。红景天含有黑景天、酪醇、没食子酸等化合物，具有"活气促血、清脉、镇静哮喘"等功能，用于治疗缺氧相关疾病8,9,10,11。野外调查表明，在海拔4,000-5,600 m的高山距骨带、沟壑斜坡和岩石缝隙中可以找到红景天。它的生长环境寒冷，阳光充足，辐射强烈，属于高寒草甸生态系统。红景天根据生长地形可分布在层状和点状种群中，通过异花授粉进行基因流动。
红景天属的花粉流产、非法挖掘和生态环境的退化使红景天成为濒危物种6,12。鉴于红景天的高药用价值，预计假冒产品将流入市场。本文介绍了红景天的栖息地和一些方便的实验室鉴定方法。首先，我们观察了红景天的生长环境及其药用特性。其次，通过显微镜观察药粉的微观结构;最后一步是关键点。根据红景天在某种物质中的不同吸附或溶解特性，对红景天的代表性成分进行分离和鉴定。基于DNA的药用植物鉴定或代谢组学分析方法复杂且昂贵13.这些基本、方便、经济的方法可以快速识别红景天。

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			<td>0.22 &#956;m millipore filter</td>
			<td>Millipore</td>
			<td>SLGP033RB</td>
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			<td>Automatic sampling machine</td>
			<td>CAMAG</td>
			<td>ATS 4</td>
			<td></td>
		</tr>
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			<td>Chloral hydrate</td>
			<td>Fuzhou Brunei Technology Co., Ltd</td>
			<td>ST1002</td>
			<td></td>
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			<td>Chromatographic&#160;sample&#160;bottles</td>
			<td>Zhejiang ALWSCI Technology Co., Ltd</td>
			<td>C0000008</td>
			<td></td>
		</tr>
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			<td>Conical flask</td>
			<td>Sichuan Shubo Co., Ltd</td>
			<td>1121</td>
			<td></td>
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			<td>Cover glass</td>
			<td>Citotest Labware Manufacturing Co., Ltd</td>
			<td>10211818c</td>
			<td></td>
		</tr>
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			<td>Dissecting needle</td>
			<td>Shanghai Bingyu Fluid Technology Co., Ltd</td>
			<td>BY-5026</td>
			<td></td>
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			<td>Electronic balance</td>
			<td>SHIMADZU</td>
			<td>ATX124</td>
			<td></td>
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			<td>Ethyl acetate</td>
			<td>Chengdu Kelong Chemical Co., Ltd</td>
			<td>2022120901</td>
			<td></td>
		</tr>
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			<td>Formic acid</td>
			<td>Chengdu Kelong Chemical Co., Ltd</td>
			<td>2021110801</td>
			<td></td>
		</tr>
		<tr>
			<td>Gallic acid</td>
			<td>Chengdu Herbpurify Co., Ltd</td>
			<td>M-017</td>
			<td></td>
		</tr>
		<tr>
			<td>Glycerol</td>
			<td>Sinopharm Chemical Reagent Co., Ltd</td>
			<td>10010618</td>
			<td></td>
		</tr>
		<tr>
			<td>High speed&#160; crusher</td>
			<td>Beijing Zhongxingweiye Instrument Co., Ltd</td>
			<td>FW-100</td>
			<td></td>
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			<td>Methanol</td>
			<td>Chengdu Kelong Chemical Co., Ltd</td>
			<td>20230108</td>
			<td></td>
		</tr>
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			<td>Microscope</td>
			<td>Chongqing Oprec Nistrument Co.,&#160; Ltd</td>
			<td>B203</td>
			<td></td>
		</tr>
		<tr>
			<td>Microscope slide</td>
			<td>Citotest Labware Manufacturing Co., Ltd</td>
			<td>7105P-G</td>
			<td></td>
		</tr>
		<tr>
			<td>Oven</td>
			<td>Shanghai Yuejin Medical Equipment Co., Ltd</td>
			<td>DHG-8145</td>
			<td></td>
		</tr>
		<tr>
			<td>Pharmacopoeia sieve</td>
			<td>Hangzhou Xingrun sieve factory</td>
			<td>572423281330</td>
			<td></td>
		</tr>
		<tr>
			<td>Pipette</td>
			<td>Changde BKMAM Biotechnology Co., Ltd</td>
			<td>120302008</td>
			<td></td>
		</tr>
		<tr>
			<td>Salidroside</td>
			<td>Chengdu Herbpurify Co., Ltd</td>
			<td>H-040</td>
			<td></td>
		</tr>
		<tr>
			<td>Saturate tank&#160;</td>
			<td>Yancheng Liegu Technology Co., Ltd</td>
			<td>10*20 P-1</td>
			<td></td>
		</tr>
		<tr>
			<td>Silica gel plate</td>
			<td>Yantai Jiangyou Silica Gel Development&#160; Co., Ltd</td>
			<td>HSG20211227</td>
			<td></td>
		</tr>
		<tr>
			<td>Trichloromethane</td>
			<td>Chengdu Kelong Chemical Co., Ltd</td>
			<td>20221013-1</td>
			<td></td>
		</tr>
		<tr>
			<td>Tweezer&#160;</td>
			<td>Changde BKMAM Biotechnology Co., Ltd</td>
			<td>130302027</td>
			<td></td>
		</tr>
		<tr>
			<td>Tyrosol</td>
			<td>Chengdu Herbpurify Co., Ltd</td>
			<td>L-042</td>
			<td></td>
		</tr>
		<tr>
			<td>Ultrasound equipment</td>
			<td>Ningbo Xinyi Ultrasonic Equipment Co., Ltd</td>
			<td>SB-8200DTS</td>
			<td></td>
		</tr>
		<tr>
			<td>Volumetric pipet</td>
			<td>Changde BKMAM Biotechnology Co., Ltd</td>
			<td>120301006</td>
			<td></td>
		</tr>
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</table>

field collection and laboratory routine identification of rhodiola crenulata

药材鉴定是药品安全的前提和保障。大多数科研人员必然会青睐简单、快速、有效、廉价的草药鉴定过程。 红景天 是一种生长在高海拔地区的传统藏药，主要分布于中国西藏、云南和四川地区。 红景天 具有抗炎、抗缺氧、抗氧化等多种生物活性，具有很大的发展潜力。随着市场需求的增加和资源含量的迅速减少，大量混淆的 红景天 产品一直困扰着人们。因此，该协议引入了在现场鉴定 红景天 的标准过程，并结合常规实验室测试。生境、微观特征和薄层色谱法的结合，无疑将快速、高效、经济地鉴定 红景 天，为藏医药的不断发展和药材的质量控制做出贡献。

该实验方案描述了在田间识别和收集 红景天 。 红景天 倾向于生活在高山距骨带、沟壑斜坡和高海拔的岩石缝隙中。 红景天 的生境、全株、花和叶如 图2所示。 红景天 具有红褐色的根茎（图3A）。药粉的代表性图像如 图3B所示。根据上述实验方案，显微结果可列举如下:1）棕黄色或无色软木细胞，外?...

Watch this Scientific Journal Video about Field Collection and Laboratory Routine Identification of Rhodiola crenulata at JoVE.com

作者聚焦:开发 红景天 鉴定方法并研究其资源分布和药理作用

在这里，我们从栖息地、植物形态、药用特性、微观特征和薄层色谱法中描述了红 景天 的鉴定。

红景天的现场采集和实验室常规鉴定

The identification of medicinal materials is the premise and guarantee of drug safety. The majority of scientific researchers are bound to favor the ...

我们的研究范围集中在进行民族医学的资源调查。具体而言，本研究旨在开发一种快速方便的红景天鉴定方法。我们提供的方案提供了一种具有成本效益、直接和快速的方法来识别各种药材。我们的实验室将把未来的研究工作集中在两个主要领域的调查上，即与红景天相关的研究分布和该物种的药理学影响。

field-collection-laboratory-routine-identification-rhodiola

Research

JoVE Journal

Medicine

Field Collection and Laboratory Routine Identification of Rhodiola crenulata

545 次观看.  Chengdu University of Traditional Chinese Medicine. 我们的研究范围集中在进行民族医学的资源调查。具体而言，本研究旨在开发一种快速方便的红景天鉴定方法。我们提供的方案提供了一种具有成本效益、直接和快速的方法来识别各种药材。我们的实验室将把未来的研究工作集中在两个主要领域的调查上，即与红景天相关的研究分布和该物种的药理学影响。

视频: 作者聚焦:开发 红景天 鉴定方法并研究其资源分布和药理作用

Collection Protocol for Human Pancreas

This dissection and sampling procedure was developed for the Network for Pancreatic Organ Donors with Diabetes (nPOD) program to standardize preparation of pancreas recovered from cadaveric organ donors. The pancreas is divided into 3 main regions (head, body, tail) followed by serial transverse sections throughout the medial to lateral axis. Alternating sections are used for fixed paraffin and fresh frozen blocks and remnant samples are minced for snap frozen sample preparations, either with or without RNAse inhibitors, for DNA, RNA, or protein isolation. The overall goal of the pancreas dissection procedure is to sample the entire pancreas while maintaining anatomical orientation.
Endocrine cell heterogeneity in terms of islet composition, size, and numbers is reported for human islets compared to rodent islets 1. The majority of human islets from the pancreas head, body and tail regions are composed of insulin-containing &beta;-cells followed by lower proportions of glucagon-containing &alpha;-cells and somatostatin-containing &delta;-cells. Pancreatic polypeptide-containing PP cells and ghrelin-containing epsilon cells are also present but in small numbers. In contrast, the uncinate region contains islets that are primarily composed of pancreatic polypeptide-containing PP cells 2. These regional islet variations arise from developmental differences. The pancreas develops from the ventral and dorsal pancreatic buds in the foregut and after rotation of the stomach and duodenum, the ventral lobe moves and fuses with the dorsal 3. The ventral lobe forms the posterior portion of the head including the uncinate process while the dorsal lobe gives rise to the rest of the organ. Regional pancreatic variation is also reported with the tail region having higher islet density compared to other regions and the dorsal lobe-derived components undergoing selective atrophy in type 1 diabetes4,5.
Additional organs and tissues are often recovered from the organ donors and include pancreatic lymph nodes, spleen and non-pancreatic lymph nodes. These samples are recovered with similar formats as for the pancreas with the addition of isolation of cryopreserved cells. When the proximal duodenum is included with the pancreas, duodenal mucosa may be collected for paraffin and frozen blocks and minced snap frozen preparations.

This video demonstrates a dissection procedure for processing human pancreas into multiple storage formats. Anatomical orientation is maintained throughout the pancreatic regions to allow definition of regional islet composition and density.

人体胰腺的收集协议

This dissection and sampling procedure was developed for the Network for Pancreatic Organ Donors with Diabetes (nPOD) program to standardize preparation ...

科研

医学

The Use of Primary Human Fibroblasts for Monitoring Mitochondrial Phenotypes in the Field of Parkinson's Disease

Parkinson's disease (PD) is the second most common movement disorder and affects 1% of people over the age of 60 1. Because ageing is the most important risk factor, cases of PD will increase during the next decades 2. Next to pathological protein folding and impaired protein degradation pathways, alterations of mitochondrial function and morphology were pointed out as further hallmark of neurodegeneration in PD 3-11.
After years of research in murine and human cancer cells as in vitro models to dissect molecular pathways of Parkinsonism, the use of human fibroblasts from patients and appropriate controls as ex vivo models has become a valuable research tool, if potential caveats are considered. Other than immortalized, rather artificial cell models, primary fibroblasts from patients carrying disease-associated mutations apparently reflect important pathological features of the human disease.
Here we delineate the procedure of taking skin biopsies, culturing human fibroblasts and using detailed protocols for essential microscopic techniques to define mitochondrial phenotypes. These were used to investigate different features associated with PD that are relevant to mitochondrial function and dynamics. Ex vivo, mitochondria can be analyzed in terms of their function, morphology, colocalization with lysosomes (the organelles degrading dysfunctional mitochondria) and degradation via the lysosomal pathway. These phenotypes are highly relevant for the identification of early signs of PD and may precede clinical motor symptoms in human disease-gene carriers. Hence, the assays presented here can be utilized as valuable tools to identify pathological features of neurodegeneration and help to define new therapeutic strategies in PD.

Fibroblasts from patients carrying mutations in Parkinson's disease-causing genes represent an easily accessible ex vivo model to study disease-associated phenotypes. Live cell imaging gives the opportunity to study morphological and functional parameters in living cells. Here we describe the preparation of human fibroblasts and subsequent monitoring of mitochondrial phenotypes.

利用人类纤维原细胞线粒体表型的监测领域的帕金森氏病

Parkinson's disease (PD) is the second most  common movement disorder and affects 1% of people over the age of 60 1. Because ageing is  the most important ...

Collection and Extraction of Saliva DNA for Next Generation Sequencing

The preferred source of DNA in human genetics research is blood, or cell lines derived from blood, as these sources yield large quantities of high quality DNA. However, DNA extraction from saliva can yield high quality DNA with little to no degradation/fragmentation that is suitable for a variety of DNA assays without the expense of a phlebotomist and can even be acquired through the mail. However, at present, no saliva DNA collection/extraction protocols for next generation sequencing have been presented in the literature. This protocol optimizes parameters of saliva collection/storage and DNA extraction to be of sufficient quality and quantity for DNA assays with the highest standards, including microarray genotyping and next generation sequencing.

DNA extraction from saliva can provide a readily available source of high molecular weight DNA, with little to no degradation/fragmentation. This protocol provides optimized parameters for saliva collection/storage and DNA extraction to be of sufficient quality and quantity for downstream DNA assays with high quality requirements.

收集和唾液DNA提取用于新一代测序

The preferred source of DNA in human genetics research is blood, or cell lines derived from blood, as these sources yield large quantities of high quality ...

Collection, Isolation, and Flow Cytometric Analysis of Human Endocervical Samples

Despite the public health importance of mucosal pathogens (including HIV), relatively little is known about mucosal immunity, particularly at the female genital tract (FGT). Because heterosexual transmission now represents the dominant mechanism of HIV transmission, and given the continual spread of sexually transmitted infections (STIs), it is critical to understand the interplay between host and pathogen at the genital mucosa. The substantial gaps in knowledge around FGT immunity are partially due to the difficulty in successfully collecting and processing mucosal samples. In order to facilitate studies with sufficient sample size, collection techniques must be minimally invasive and efficient. To this end, a protocol for the collection of cervical cytobrush samples and subsequent isolation of cervical mononuclear cells (CMC) has been optimized. Using ex vivo flow cytometry-based immunophenotyping, it is possible to accurately and reliably quantify CMC lymphocyte/monocyte population frequencies and phenotypes. This technique can be coupled with the collection of cervical-vaginal lavage (CVL), which contains soluble immune mediators including cytokines, chemokines and anti-proteases, all of which can be used to determine the anti- or pro-inflammatory environment in the vagina.

The use of cytobrush sampling to collect lymphocytes and monocytes from the endocervix is a minimally invasive technique that provides samples for analysis of female genital tract immunity. In this protocol, we describe the collection of cytobrush samples and immune cell isolation for flow cytometry assays.

收集，分离，和人类宫颈样品的流式细胞仪分析

Despite the public health importance of mucosal pathogens (including HIV), relatively little is known about mucosal immunity, particularly at the female ...

Localization, Identification, and Excision of Murine Adipose Depots

Obesity has increased dramatically in the last few decades and affects over one third of the adult US population. The economic effect of obesity in 2005 reached a staggering sum of $190.2 billion in direct medical costs alone. Obesity is a major risk factor for a wide host of diseases. Historically, little was known regarding adipose and its major and essential functions in the body. Brown and white adipose are the two main types of adipose but current literature has identified a new type of fat called brite or beige adipose. Research has shown that adipose depots have specific metabolic profiles and certain depots allow for a propensity for obesity and other related disorders. The goal of this protocol is to provide researchers the capacity to identify and excise adipose depots that will allow for the analysis of different factorial effects on adipose; as well as the beneficial or detrimental role adipose plays in disease and overall health. Isolation and excision of adipose depots allows investigators to look at gross morphological changes as well as histological changes. The adipose isolated can also be used for molecular studies to evaluate transcriptional and translational change or for in vitro experimentation to discover targets of interest and mechanisms of action. This technique is superior to other published techniques due to the design allowing for isolation of multiple depots with simplicity and minimal contamination.

Due to the drastic and negative connection between obesity and other comorbidities, research on the role adipose plays in disease and overall health is warranted. We present a protocol for the isolation and excision of adipose depots allowing for the study of adipose using in situ and in vitro methods.

本地化识别，切除脂肪小鼠油库，并

Obesity has increased dramatically in the last few decades and affects over one third of the adult US population. The economic effect of obesity in 2005 ...

Methodology for Sputum Induction and Laboratory Processing

The technique of sputum induction and processing is a recognized non-invasive method allowing the collection and analysis of cells from the airways, which is interesting in various respiratory diseases like asthma, chronic obstructive pulmonary disease (COPD), chronic cough, or idiopathic pulmonary fibrosis. This technique is well tolerated, safe and non-invasive, but is currently limited to research services and specialized centers in clinical practice because it is technically demanding, time-consuming, and requires trained staff. The success rate of sputum induction and analysis is about 80%.
Here, we describe the induction and laboratory processing of sputum samples. Sputum is induced by inhalation of hypertonic or isotonic saline with salbutamol. For the processing, we use the whole sputum technique. Dithiothreitol (DTT) is used to allow mucolysis of sputum samples. The primary aim of sputum processing is to obtain a differential cell count to study the cell types present in the airway lumen. Additional analyses may also be performed on sputum supernatant and sputum cells, which may allow further investigation into inflammatory processes and immune mechanisms. Examples include studying mediators in sputum supernatant and performing a large spectrum of analysis on sputum cells such as flow cytometry, genomics, or proteomics.
Finally, representative results of sputum analysis in healthy controls, asthmatics, and COPD patients are presented.

Here we describe the technique of sputum induction. This protocol also explains the sputum processing to perform a differential cell count and to collect sputum supernatant and cells for further analysis.

痰液诱导和实验室处理方法

The technique of sputum induction and processing is a recognized non-invasive method allowing the collection and analysis of cells from the airways, which ...

Routine Screening Method for Microparticles in Platelet Transfusions

Platelet inventory management based on screening microparticle content in platelet concentrates is a new quality improvement initiative for hospital blood banks. Cells fragment off microparticles (MP) when they are stressed. Blood and blood components may contain cellular fragments from a variety of cells, most notably from activated platelets. When performing their roles as innate immune cells and major players in coagulation and hemostasis, platelets change shape and generate microparticles. With dynamic light scattering (DLS)-based microparticle detection, it is possible to differentiate activated (high microparticle) from non-activated (low microparticle) platelets in transfusions, and optimize the use of this scarce blood product. Previous research suggests that providing non-activated platelets for prophylactic use in hematology-oncology patients could reduce their risk of becoming refractory and improve patient care. The goal of this screening method is to routinely differentiate activated from non-activated platelets. The method described here outlines the steps to be performed for routine platelet inventory management in a hospital blood bank: obtaining a sample from a platelet transfusion, loading the sample into the capillary for DLS measurement, performing the DLS test to identify microparticles, and using the reported microparticle content to identify activated platelets.

Platelet inventory management based on screening microparticle content in platelet concentrates is a new quality improvement initiative in hospital blood banks. The goal is to differentiate activated from non-activated platelets to optimize the use of platelets. Providing non-activated platelets to hematology-oncology patients might reduce their high risk to become refractory.

血小板输血中微粒的常规筛选方法

Platelet inventory management based on screening microparticle content in platelet concentrates is a new quality improvement initiative for hospital blood ...

Remote Laboratory Management: Respiratory Virus Diagnostics

An uptick in recent pandemics (Ebola, Zika, MERS, influenza, etc.) underlines the need for a more 'nimble,' coordinated response that addresses a multitude of issues ranging from transportation, access, facilities, equipment, and communication to provider training. To address this need, we have developed an innovative, scalable, logistics-enhanced, mobile, laboratory facility for emergencies and epidemics in resource-constrained global settings. Utilizing a background in clinical operations as an academic medical center, we designed a rapidly-deployable, modular BSL-2 and BSL-3 facility with user-friendly software for tracking and management of drugs and supplies in remote regions during epidemics and outbreaks. Here, we present our intermodal, mobile, expandable shipping-container laboratory units. The design of the laboratory facilitates off-grid usage by minimizing power consumption and allowing alternate water sources. The unit's information communication technology (ICT) platform provides (i) user-friendly tablet-based documentation, (ii) enhanced tracking of patients and supplies, and (iii) integrated communication onsite with built-in telehealth capabilities. To ensure quality in remote environments, we have developed a checklist for a basic laboratory workflow and a protocol for respiratory viral diagnosis using reverse-transcription polymerase chain reaction (RT-PCR). As described, this innovative and comprehensive approach allows for the provision of laboratory capability in resource-limited global environments.

A rapidly-deployable, off-grid laboratory has been designed and built for remote, resource-constrained global settings. The features and critical aspects of the logistically-enhanced, expandable, multifunctional laboratory modules are explored. A checklist for a basic laboratory workflow and a protocol for a respiratory viral diagnostic test are developed and presented.

远程实验室管理: 呼吸道病毒诊断

An uptick in recent pandemics (Ebola, Zika, MERS, influenza, etc.) underlines the need for a more 'nimble,' coordinated response that addresses a ...

Learning Modern Laryngeal Surgery in a Dissection Laboratory

Surgery for laryngeal malignancies requires millimetric accuracy from the different endoscopic and open techniques available. Practice of this surgery is almost completely reserved to a few referral centers that deal with a large proportion of this pathology. Practice on human specimens is not always possible for ethical, economic, or availability reasons. The aim of this study is to provide a reproducible method for the organization of a laryngeal laboratory on ex vivo animal models where it is possible to approach, learn, and refine laryngeal techniques. Porcine and ovine larynges are ideal, affordable, models to simulate laryngeal surgery given their similarity to the human larynx in their anatomical layout and tissue composition. Herein, the surgical steps of transoral laser surgery, open partial horizontal laryngectomy, and total laryngectomy are reported. The merging of endoscopic and exoscopic views guarantees an inside-out perspective, which is vital for the comprehension of the complex laryngeal anatomy. The method was successfully adopted during three sessions of a dissection course &#34;Lary-Gym&#34;. Further perspectives on robotic surgical training are described.

The purpose of this paper is to illustrate how to organize a reproducible laboratory for laryngeal surgery on affordable and closely similar animal laryngeal models in order to improve anatomical and surgical knowledge and skills.

在解剖实验室学习现代喉外科

Surgery for laryngeal malignancies requires millimetric accuracy from the different endoscopic and open techniques available. Practice of this surgery is ...

Virtual Reality Tools for Assessing Unilateral Spatial Neglect: A Novel Opportunity for Data Collection

Unilateral spatial neglect (USN) is a syndrome characterized by inattention to or inaction in one side of space and affects between 23-46% of acute stroke survivors. The diagnosis and characterization of these symptoms in individual patients can be challenging and often requires skilled clinical staff. Virtual reality (VR) presents an opportunity to develop novel assessment tools for patients with USN.
We aimed to design and build a VR tool to detect and characterize subtle USN symptoms, and to test the tool on subjects treated with inhibitory repetitive transcranial magnetic stimulation (TMS) of cortical regions associated with USN.
We created three experimental conditions by applying TMS to two distinct regions of cortex associated with visuospatial processing- the superior temporal gyrus (STG) and the supramarginal gyrus (SMG) - and applied sham TMS as a control. We then placed subjects in a virtual reality environment in which they were asked to identify the flowers with lateral asymmetries of flowers distributed across bushes in both hemispaces, with dynamic difficulty adjustment based on each subject's performance. 
We found significant differences in average head yaw between subjects stimulated at the STG and those stimulated at the SMG and marginally significant effects in the average visual axis.
VR technology is becoming more accessible, affordable, and robust, presenting an exciting opportunity to create useful and novel game-like tools. In conjunction with TMS, these tools could be used to study specific, isolated, artificial neurological deficits in healthy subjects, informing the creation of VR-based diagnostic tools for patients with deficits due to acquired brain injury. This study is the first to our knowledge in which artificially generated USN symptoms have been evaluated with a VR task.

The goal was to design, build, and pilot a novel virtual reality task to detect and characterize unilateral spatial neglect, a syndrome affecting 23-46% of acute stroke survivors, expanding the role of virtual reality in the study and management of neurologic disease.

用于评估单边空间忽视的虚拟现实工具:数据收集的新机遇

Unilateral spatial neglect (USN) is a syndrome characterized by inattention to or inaction in one side of space and affects between 23-46% of acute stroke ...