Name: basophil activation test for allergy diagnosis
Uploaded: 2021-05-31T14:00:00
Description: Watch this Scientific Journal Video about Basophil Activation Test for Allergy Diagnosis at JoVE.com

我们感谢克劳迪娅·科拉扎（Claudia Corazza）宝贵的英语语言支持。这项工作得到了MIENCO卫生研究所“Carlos III”（ISCIII）的支持（由ERDF共同资助的赠款：“Una manera de hacer Europa”;授权号PI20/01715;PI18/00095;PI17/01410;PI17/01318;PI17/01237 和 RETIC ARADYAL RD16/0006/0001;安达卢西亚地区卫生部（批准号PI-0127-2020，PIO-0176-2018;PE-0172-2018;PE-0039-2018;PC-0098-2017;PI-0075-2017;PI-0241-2016）。ID是一名临床研究者（B-0001-2017），AA持有高级博士后合同（RH-0099-2020），两者都由安达卢西亚地区卫生部支持（由ESF共同资助：“Andalucía se mueve con Europa”）。

协议执行是根据赫尔辛基原则宣言进行的，并得到当地道德委员会（西班牙马拉加省调查委员会）的批准。所有受试者都口头告知了这项研究，并签署了相应的知情同意书。
注意：本协议详细介绍了作者每天使用的BAT程序。然而，这不是一种标准化的方法，并且与其他作者发表的程序存在差异。主要的方案修改与刺激缓冲液中IL-3的使用，刺激物的孵育时间，停止嗜碱?...

BAT是一种补充体 外 诊断试验，用于评估IgE介导的过敏反应，已被证明可用于诊断由药物，食物或吸入剂等不同触发因素引起的反应，以及某些形式的慢性荨麻疹。一般来说，如果 i） 变应原/药物在 ST 中产生假阳性结果;ii） 过敏原/药物不能用于 ST 或 sIgE 定量;iii） 临床病史与 ST 或 sIgE 测定之间存在不一致;iv）症状表明ST段可能诱发全身反应;v） 在 CCT 之前，用于致病原原/药物确认<sup class...

IgE 介导的过敏诊断基于临床病史、皮肤试验 （ST）、血清特异性 IgE （sIgE） 的定量，以及对照激发试验 （CCT）（如果需要和指征）1，2，3，4，5，6。然而，临床病史可能不可靠，因为可能缺乏准确的信息，ST 和 CCT 不是无风险的程序，在经历严重危及生命的反应的受试者中是禁忌的 1，2，3，4，5，6.这些问题，加上通过经过验证和商业的荧光酶免疫测定测定sIgE仅适用于少数过敏原和药物的事实，突出了其他体外功能测定（如嗜碱性粒细胞活化试验（BAT））的重要作用。
嗜碱性粒细胞是参与 IgE 介导的过敏反应的关键效应细胞，在过敏原/药物暴露后，在细胞表面高亲和力受体 （FcεRI） 上结合的相邻 sIgE 交联时被激活。嗜碱性粒细胞活化触发细胞脱颗粒和细胞胞浆内分泌颗粒中所含的预形成和新合成的炎症介质的释放7，8，9。BAT是一种体外方法，它试图在刺激（过敏原或药物）存在的情况下模拟嗜碱性粒细胞活化，并通过流式细胞术7，10确定嗜碱性粒细胞活化标志物表达的变化。有不同的策略来鉴定嗜碱性粒细胞（IgE+、CCR3+、CRTH2+、CD203c+）和使用荧光染料标记抗体的组合来测量细胞活化（主要是 CD63 和 CD203c 的上调7，10）。CD63是临床验证的最佳活化标志物11，12，13，14，是一种膜蛋白，锚定在含有组胺的分泌颗粒上，在细胞活化和颗粒与膜融合后，在嗜碱性粒细胞表面表达15，16，17，18，19，20，21.CD203c是一种表面标志物，在嗜碱性粒细胞上形成型表达，并在FcεRI刺激后上调，在BAT15，22，23，24，25中也显示出可靠的结果。此外，它似乎与CD6326共同表达。
在过去的几十年中，BAT已被证明可用于诊断由药物，食物或吸入剂等不同触发因素引起的IgE介导的过敏反应，以及某些形式的慢性荨麻疹，如下所述。然而，该技术在诊断算法中的位置变化很大，并且尚未很好地确定。
药物超敏反应 BAT已被证明可作为特定药物和患者的补充测试，特别是对于那些由于ST的诊断价值尚未很好地确定而出现严重反应的患者，因为它们对有限数量的药物进行了验证和标准化27，28，29，30。此外，sIgE 的定量仅适用于有限数量的药物，灵敏度低于 ST27，28，29，30，31，32。因此，药物超敏反应的诊断通常依赖于药物激发试验，这可能在经历严重危及生命的反应的受试者中禁忌33。
据报道，在选定的患者中使用BAT有希望的结果，这些患者报告对β内酰胺类（BLs）的即时超敏反应20，34，35，36，37，38，39，神经肌肉阻滞剂（NMBAs）19，22，40，41，42，43，44，45，氟喹诺酮类（FQ）46，47，48，49，吡唑啉酮类50，51，52，放射造影剂（RCM）53，54，55，56和铂化合物57，58，59.据报道，BAT的敏感性和特异性分别为51.7-66.9%和89.2-97.8%;阳性和阴性预测值分别在 93.4% 和 66.3% 之间27，31。此外，BAT已被提议作为铂化合物脱敏过程中突破性反应的预测生物标志物，因为在药物脱敏期间不良反应风险高的患者中，CD203c表达与CD63相比增加57。
值得注意的是，BAT仅在反应涉及嗜碱性粒细胞脱颗粒时才对药物超敏反应有用;因此，它在由酶促抑制环加氧酶 142引起的反应中没有用。
食物过敏 BAT已成为食物过敏的潜在诊断工具，因为测定血清sIgE对整个过敏原提取物或单一过敏原通常是模棱两可的，需要口服食物激发试验来确认诊断，这与药物超敏反应类似，是一种昂贵且并非无风险的过程60。多项研究显示了牛奶 61，62、鸡蛋 61，63、小麦64、65、66、67、68、花生 63、69、70、71、72、榛子73、74、75，76，77，贝类78，桃子79，80，81，苹果21，芹菜和胡萝卜82，83。
与血清中的STs和sIgE相比，BAT在食物过敏诊断中的主要附加值是它显示出更高的特异性和相似的敏感性。因此，BAT是区分临床过敏患者与具有高特异性（75-100%）和敏感性（77-98%）的致敏但耐受的受试者的有用工具63，69，84。敏感性和特异性值取决于变应原和其他因素，如表型（例如口腔过敏综合征与全身过敏反应）、年龄和地理相关的致敏模式63，85。
使用单一过敏原成分的BAT可以潜在地提高某些食物过敏原的诊断准确性61，80。有使用种子储存蛋白的研究（例如，来自花生的Ara h 1，Ara h 2，Ara h 3和Ara h 6）86;脂质转移蛋白（例如，来自桃子的Pru p 3和来自花生的Ara h 9）80，86;和 Bet v 1 同系物（例如，来自花生的 Ara h 8）87。其他潜在的效用与在花粉食物过敏综合征21，87，88，对红肉过敏89或食物依赖性运动诱导的过敏反应66的情况下识别罪魁祸首过敏原有关。
有趣的是，BAT可以提供有关过敏反应的严重程度和阈值的信息，因为反应更严重的患者表现出更大比例的活化嗜碱性粒细胞，如花生和牛奶过敏患者的研究中观察到的那样84，90，91;对微量过敏原有反应的患者表现出更大的嗜碱性粒细胞敏感性84，90，92。这些数据表明，BAT可能有助于识别需要更密切随访和更强化教育的高风险过敏患者93。此外，据报道，BAT可以预测食物挑战反应70，91，92，94和反应性阈值90，95，以帮助确定何时可以安全地（重新）引入食物84。然而，这些发现在一些研究中存在争议63，96，需要更多的研究。
另一方面，随着时间的推移，BAT已被用于监测食物过敏的缓解情况，无论是自然的还是在免疫调节治疗下的，到目前为止，仅通过口服食物激发进行评估，具有相关的风险和成本84，97，98，99，100，101，102，103，104，105，106，107，108。此外，它还被用于监测奥马珠单抗在食物过敏中的作用，因为在奥马珠单抗治疗期间嗜碱性粒细胞活化降低，但在停止治疗后增加109。
吸入剂过敏 BAT很少对吸入剂过敏有益，因为可以通过sIgE定量和ST常规确定诊断。然而，在局部过敏性鼻炎病例（sIgE 水平检测不到和鼻激发试验阳性的 ST 阴性）病例中，BAT 允许诊断 50% 的病例110.进一步报道了嗜碱性粒细胞敏感性与鼻/支气管激发试验反应之间的相关性，以及哮喘严重程度与奥马珠单抗 111，112 治疗效果之间的相关性。
BAT也被用于监测屋尘螨和花粉的变应原免疫治疗，因为在免疫治疗期间嗜碱性粒细胞敏感性降低，可能是由于阻断IgG抗体113，114，115，116，117的干扰。
膜翅目毒液过敏 膜翅目毒液过敏的诊断通常基于 ST 和血清 sIgE。BAT显示出高敏感性（85-100%）和特异性（83-100%），据报道，对于结果模棱两可的病例或有提示性毒液过敏临床病史但sIgE检测不到且ST 118，119阴性的患者，BAT是有用的。然而，BAT似乎不能预测这些反应的严重程度120，121。
高达 60% 的患者对黄蜂和蜂毒均表现出 sIgE，鉴定显性变应原对于充分的免疫治疗至关重要。据报道，在这些情况下，BAT可用于鉴定主要过敏原119，122，123，124。尽管对蜜蜂和黄蜂毒液的主要过敏原的sIgE可能会降低BAT在两种毒液双重阳性患者中的效用，但它主要在sIgE测定结果为阴性的受试者中提供有用的信息123。
一些研究表明，在毒液免疫治疗的积累阶段，BAT可能可作为副作用的预测生物标志物，因为据报道这种治疗选择会降低嗜碱性粒细胞的敏感性。然而，反应性并没有降低，这个BAT实用程序现在有争议的是13，120，125，126，127，128，129，130。
荨麻疹和血管性水肿 一部分慢性荨麻疹患者具有自身免疫病理生理学，这是由于针对自身过敏原的 IgE 自身抗体和靶向肥大细胞表面存在的 FcεRI 或 IgE-FcεRI 复合物的 IgG 自身抗体 131，132。在临床实践中，这种类型的慢性荨麻疹的诊断依赖于自体血清ST阳性，该血清ST具有意外感染的风险。BAT已被提议作为诊断和监测疑似慢性荨麻疹患者的体外检查。据报道，在慢性荨麻疹患者的血清刺激后，嗜碱性粒细胞表面的CD63和CD203c表达均增加，显示检测到活性自身抗体133，134，135，136，137。最近，据报道，与BAT138阴性的患者相比，BAT阳性患者通常经历最活跃的疾病状态，通过荨麻疹活动评分评估，并且需要更高剂量的抗组胺药联合三线治疗（环孢素A或奥马珠单抗）。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>5 mL Round Bottom Polystyrene Test Tube, without Cap, Nonsterile</td>
			<td>Corning</td>
			<td>352008</td>
			<td></td>
		</tr>
		<tr>
			<td>APC anti-human CD193 (CCR3) Antibody</td>
			<td>BioLegend</td>
			<td>310708</td>
			<td></td>
		</tr>
		<tr>
			<td>BD FACSCalibur Flow Cytometer</td>
			<td>BD Biosciences</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Calcium chloride</td>
			<td>Sigma-Aldrich</td>
			<td>C1016</td>
			<td></td>
		</tr>
		<tr>
			<td>FITC anti-human CD63 Antibody</td>
			<td>BioLegend</td>
			<td>353006</td>
			<td></td>
		</tr>
		<tr>
			<td>HEPES (1 M)</td>
			<td>Thermo-Fisher</td>
			<td>15630106</td>
			<td></td>
		</tr>
		<tr>
			<td>Lysing Solution 10x concentrated</td>
			<td>BD Biosciences</td>
			<td>349202</td>
			<td></td>
		</tr>
		<tr>
			<td>Magnesium chloride</td>
			<td>Sigma-Aldrich</td>
			<td>M8266</td>
			<td></td>
		</tr>
		<tr>
			<td>N-Formyl-Met-Leu-Phe</td>
			<td>Sigma-Aldrich</td>
			<td>F3506</td>
			<td></td>
		</tr>
		<tr>
			<td>PE anti-human CD203c (E-NPP3) Antibody</td>
			<td>BioLegend</td>
			<td>324606</td>
			<td></td>
		</tr>
		<tr>
			<td>Potassium chloride</td>
			<td>Sigma-Aldrich</td>
			<td>P9541</td>
			<td></td>
		</tr>
		<tr>
			<td>Purified Mouse Anti-Human IgE</td>
			<td>BD Biosciences</td>
			<td>555857</td>
			<td></td>
		</tr>
		<tr>
			<td>Recombinant Human IL-3</td>
			<td>R&#38;D Systems</td>
			<td>203-IL</td>
			<td></td>
		</tr>
		<tr>
			<td>Sheath Fluid</td>
			<td>BD Biosciences</td>
			<td>342003</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium chloride</td>
			<td>Sigma-Aldrich</td>
			<td>S3014</td>
			<td></td>
		</tr>
		<tr>
			<td>TUBE 9 mL LH Lithium Heparin</td>
			<td>Greiner Bio-One</td>
			<td>455084</td>
			<td></td>
		</tr>
		<tr>
			<td>Tween 20</td>
			<td>Sigma-Aldrich</td>
			<td>P1379</td>
			<td></td>
		</tr>
	</tbody>
</table>

basophil activation test for allergy diagnosis

嗜碱性粒细胞活化试验 （BAT） 是一种补充性体外诊断试验，除了临床病史、皮肤试验 （ST） 和特异性 IgE （sIgE） 测定 外 ，还可用于评估 IgE 介导的对食物、昆虫毒液、药物以及某些形式的慢性荨麻疹的过敏反应。然而，该技术在诊断算法中的作用是高度可变的，并且尚未很好地确定。
BAT基于通过流式细胞术测量活化标志物（如CD63，CD203c）来确定嗜碱性粒细胞对变应原/药物交联IgE活化的反应。该测试可以成为避免受控挑战测试以确认过敏诊断的有用和补充工具，尤其是在经历严重危及生命的反应的受试者中。一般来说，如果 i） 变应原/药物在 ST 中产生假阳性结果;ii） 没有用于 ST 或 sIgE 测定的过敏原/药物来源;iii） 患者病史与 ST 或 sIgE 测定之间存在不一致;iv）症状提示ST可能导致全身反应;v） 在考虑进行 CCT 以确认罪魁祸首过敏原/药物之前。该测试的主要局限性与非最佳灵敏度有关，特别是在药物过敏中，需要在样品提取后不超过24小时进行测试，以及实验室之间在程序，浓度和细胞标志物方面缺乏标准化。

使用变应原或药物进行的 BAT 可以检查 IgE 依赖性超敏反应。嗜碱性粒细胞反应性应在至少两个最佳浓度下测量，以获得最佳结果34 ，并通过细胞表面CD63的上调来观察活化。此外，在过敏原的情况下，为了确认嗜碱性粒细胞反应性，应通过测量多个降低的过敏原浓度的反应性来分析嗜碱性粒细胞敏感性114。该测量允许确定诱导50%嗜碱性粒细胞（EC50）反应的过敏?...

Watch this Scientific Journal Video about Basophil Activation Test for Allergy Diagnosis at JoVE.com

Basophil Activation Test for Allergy Diagnosis

嗜碱性粒细胞活化试验是一种补充性体 外 诊断试验，用于评估 IgE 介导的过敏反应，该试验基于通过流式细胞术测量活化标志物在存在特定刺激的情况下检测嗜碱性粒细胞活化。

嗜碱性粒细胞活化试验用于过敏诊断

The basophil activation test (BAT) is a complementary in vitro diagnostic test that can be used in addition to clinical history, skin test (ST), and ...

嗜碱性粒细胞活化试验是一种补充的体外诊断试验，用于评估 IgE 介导的过敏反应，这些反应已被证明对药物、食物或吸入剂引起的反应以及某些形式的慢性荨麻疹有用。该测试基于通过流式细胞术测量活化标志物来确定嗜碱性粒细胞对变应原或药物交联 IgE 活化的反应。嗜碱性粒细胞活化试验可以是一种有用且互补的工具，可以避免受控激发试验以确认过敏诊断，尤其是在经历严重危及生命的反应的受试者中。此外，与其他仅适用于少数药物和过敏原的体外方法相比，该测试的优点是能够分析对任何药物或过敏原的反应。该测试的主要局限性与非最佳敏感性有关，特别是在药物过敏中。需要在样本提取后不超过24小时进行测试，以及实验室和诊断算法之间缺乏标准化。首先在九毫升肝素化管中收集外周血，并将样品放在转子中以将其保持在室温下，直到实验方案需要为止。标记两个五毫升细胞仪管，每个管用于阴性和阳性对照。还要为每个管子标记一个用于测试的不同过敏原或药物浓度。然后将管子放在架子上，使管子完美贴合而不会滑动。对于第一个阳性对照，在0.05%PBS-T中制备N-甲酰甲硫酰基-亮酰基-苯丙氨酸或fMLP的四微摩尔溶液，以确认嗜碱性粒细胞的质量。对于第二个阳性对照，使用 PBS-T 制备每毫升 0.05 毫克的抗 IgE 溶液。通过每 20 微升刺激缓冲液中加入 CCR3-APC、CD203c-PE 和 CD63-FITC 抗体各 1 微升来制备染色混合物。然后向每个试管中加入 23 微升这种染色混合物。向阴性对照管中加入100微升PBS-T。向其中一个阳性对照管中加入 100 微升 fMLP，并在其余试管中向另一个管中加入 100 微升抗 IgE E。加入100微升不同浓度的过敏原或药物，并在37摄氏度下孵育10分钟。孵育后，向每个试管中轻轻加入100微升血液以避免溶血。然后轻轻涡旋试管，并在恒温浴中以中等搅拌在37摄氏度下孵育25分钟。将试管保持在四摄氏度至少五分钟以停止脱颗粒。向每个试管中加入两毫升裂解缓冲液以裂解红细胞。涡旋每个试管，然后在室温下孵育试管五分钟。将试管以 300 克在 4 摄氏度下离心五分钟。然后将架子倒入水槽中以倒出上清液，而细胞仍保留在管的底部。向每个试管中加入三毫升PBS-T以洗涤细胞并涡旋试管。使用相同的参数集进行第二轮离心，并通过将架子翻转到水槽中来倒出上清液。然后将样品保持在四摄氏度避光，直到流式细胞术。将流式细胞仪连接到计算机软件，等待细胞仪准备就绪。按照文本手稿中的说明加载模板和仪器设置。开始样品采集过程。要选择活化的嗜碱性粒细胞，首先，从侧散射-前向散点图中门禁淋巴细胞。然后将淋巴细胞群中的嗜碱性粒细胞作为CCR3 + CD203c +细胞进行门禁，并且每管至少获得500个嗜碱性粒细胞。使用阴性对照管将CD63阈值设置为约2.5%并分析样品。通过使用过敏原和药物进行嗜碱性粒细胞活化试验（BAT）来研究IgE依赖性超敏反应。通过测量多个降低的过敏原浓度的反应性来分析嗜碱性粒细胞敏感性，这有助于确定诱导50%嗜碱性粒细胞或EC50反应的过敏浓度。首先，从侧散射-前向散点图中对淋巴细胞进行门控。然后将嗜碱性粒细胞从淋巴细胞群中选门为CCR3 + CD203c +细胞。然后使用CCR3-CD63图分析使用CD63作为两种药物和不同浓度过敏原的激活标志物的嗜碱性粒细胞活化。该测试是用新鲜的全血进行的，必须在抽血后不超过24小时进行。测试中使用的刺激剂不应包括任何赋形剂，并且需要考虑药物的化学特性。其他附加的体外方法用于诊断IgE介导的过敏反应，或测定sIgE，或组胺释放试验。

basophil-activation-test-for-allergy-diagnosis

Research

JoVE Journal

Medicine

FISH for Pre-implantation Genetic Diagnosis

Pre-implantation genetic diagnosis (PGD) is an established alternative to pre-natal diagnosis, and involves selecting pre-implantation embryos from a cohort generated by assisted reproduction technology (ART). This selection may be required because of familial monogenic disease (e.g. cystic fibrosis), or because one partner carries a chromosome rearrangement (e.g. a two-way reciprocal translocation). PGD is available for couples who have had previous affected children, and/or in the case of chromosome rearrangements, recurrent miscarriages, or infertility. Oocytes aspirated following ovarian stimulation are fertilized by in vitro immersion in semen (IVF) or by intracytoplasmic injection of an individual spermatozoon (ICSI). Pre-implantation cleavage-stage embryos are biopsied, usually by the removal of a single cell on day 3 post-fertilization, and the biopsied cell is tested to establish the genetic status of the embryo. Fluorescence in situ hybridization (FISH) on the fixed nuclei of biopsied cells with target-specific DNA probes is the technique of choice to detect chromosome imbalance associated with chromosome rearrangements, and to select female embryos in families with X-linked disease for which there is no mutation-specific test. FISH has also been used to screen embryos for spontaneous chromosome aneuploidy (also known as PGS or PGD-AS) in order to try and improve the efficiency of assisted reproduction; however, the predictive value of this test using the spreading and FISH technique described here is likely to be unacceptably low in most people's hands and it is not recommended for routine clinical use. We describe the selection of suitable probes for single-cell FISH, spreading techniques for blastomere nuclei, and in situ hybridization and signal scoring, applied to PGD in a clinical setting.

This article describes the selection of suitable probes for single-cell FISH, spreading techniques for blastomere nuclei, and in situ hybridization and signal scoring, applied to pre-implantation genetic diagnosis (PGD) in a clinical setting.

鱼的胚胎植入前基因诊断

Pre-implantation genetic diagnosis (PGD) is an established alternative to pre-natal diagnosis, and involves selecting pre-implantation embryos from a ...

科研

医学

The Trier Social Stress Test Protocol for Inducing Psychological Stress

This article demonstrates a psychological stress protocol for use in a laboratory setting. Protocols that allow researchers to study the biological pathways of the stress response in health and disease are fundamental to the progress of research in stress and anxiety.1 Although numerous protocols exist for inducing stress response in the laboratory, many neglect to provide a naturalistic context or to incorporate aspects of social and psychological stress. Of psychological stress protocols, meta-analysis suggests that the Trier Social Stress Test (TSST) is the most useful and appropriate standardized protocol for studies of stress hormone reactivity.2 In the original description of the TSST, researchers sought to design and evaluate a procedure capable of inducing a reliable stress response in the majority of healthy volunteers.3 These researchers found elevations in heart rate, blood pressure and several endocrine stress markers in response to the TSST (a psychological stressor) compared to a saline injection (a physical stressor).3 Although the TSST has been modified to meet the needs of various research groups, it generally consists of a waiting period upon arrival, anticipatory speech preparation, speech performance, and verbal arithmetic performance periods, followed by one or more recovery periods. The TSST requires participants to prepare and deliver a speech, and verbally respond to a challenging arithmetic problem in the presence of a socially evaluative audience.3 Social evaluation and uncontrollability have been identified as key components of stress induction by the TSST.4 In use for over a decade, the goal of the TSST is to systematically induce a stress response in order to measure differences in reactivity, anxiety and activation of the hypothalamic-pituitary-adrenal (HPA) or sympathetic-adrenal-medullary (SAM) axis during the task.1 Researchers generally assess changes in self-reported anxiety, physiological measures (e.g. heart rate), and/or neuroendocrine indices (e.g. the stress hormone cortisol) in response to the TSST. Many investigators have adopted salivary sampling for stress markers such as cortisol and alpha-amylase (a marker of autonomic nervous system activation) as an alternative to blood sampling to reduce the confounding stress of blood-collection techniques. In addition to changes experienced by an individual completing the TSST, researchers can compare changes between different treatment groups (e.g. clinical versus healthy control samples) or the effectiveness of stress-reducing interventions.1

This article describes a protocol for inducing psychological stress in participants, which enables researchers to measure psychological, physiological and neuroendocrine responses to stress within single participants or between groups.

特里尔社会压力诱发心理应激的测试协议

This article demonstrates a psychological stress protocol for use in a laboratory setting.  Protocols that allow researchers to study the biological ...

Transabdominal Ultrasound for Pregnancy Diagnosis in Reeves' Muntjac Deer

Reeves' muntjac deer (Muntiacus reevesi) are a small cervid species native to southeast Asia, and are currently being investigated as a potential model of prion disease transmission and pathogenesis. Vertical transmission is an area of interest among researchers studying infectious diseases, including prion disease, and these investigations require efficient methods for evaluating the effects of maternal infection on reproductive performance. Ultrasonographic examination is a well-established tool for diagnosing pregnancy and assessing fetal health in many animal species1-7, including several species of farmed cervids8-19, however this technique has not been described in Reeves' muntjac deer. Here we describe the application of transabdominal ultrasound to detect pregnancy in muntjac does and to evaluate fetal growth and development throughout the gestational period. Using this procedure, pregnant animals were identified as early as 35 days following doe-buck pairing and this was an effective means to safely monitor the pregnancy at regular intervals. Future goals of this work will include establishing normal fetal measurement references for estimation of gestational age, determining sensitivity and specificity of the technique for diagnosing pregnancy at various stages of gestation, and identifying variations in fetal growth and development under different experimental conditions.

Transabdominal Ultrasound for Pregnancy Diagnosis in Reeves&#039; Muntjac Deer

Transabdominal ultrasound is described as an effective, noninvasive means for assessing reproductive status in Reeves' muntjac deer. These methods can be used to achieve early pregnancy diagnosis and to evaluate fetal viability. Future applications of this technique include estimation of gestational age and effects of maternal disease on fetal development.

里维斯的蒙特雅克鹿怀孕诊断的跨性别超声波

Reeves' muntjac deer (Muntiacus reevesi) are a small cervid species native to southeast Asia, and are currently being investigated as a potential model of ...

Diagnosis of Musculus Gastrocnemius Tightness - Key Factors for the Clinical Examination

Common foot and ankle pathologies have been linked to isolated Musculus gastrocnemius tightness (MGT). Various examination techniques have been described to assess MGT. Still, a standardized examination procedure is missing. Literature argues for weightbearing examination but the degree of knee flexion needed to eliminate the restraining effect of the M. gastrocnemius on ankle dorsiflexion (ADF) is unknown. This manuscript investigates the effect of knee flexion on ankle dorsiflexion and provides a detailed description of a standardized examination protocol. Examination on 20 healthy individuals revealed, that 20&#176; of knee flexion is sufficient to fully eliminate the influence of the M. gastrocnemius on ADF. This builds the prerequisite for a standardized examination for MGT. Non-weightbearing and weightbearing examination of ADF has to be conducted with the knee fully extended and at least 20&#176; flexed. Two investigators should conduct non-weightbearing testing with the subject in supine position. In order to obtain reliable results, the axis of the fibula should be marked. One examiner can conduct weightbearing examination with the subject in lunge stance. Isolated MGT is present if ADF is impaired with the knee fully extended and knee flexion results in a significant ADF increase. The herein presented standardized examination is the prerequisite for future studies aiming at establishing norm values.

Diagnosis of Musculus Gastrocnemius Tightness - Key Factors for the Clinical Examination

Isolated Musculus gastrocnemius tightness is a common cause for foot and ankle pathologies. Currently no standardized examination procedure exists. This manuscript demonstrates that 20 degree of knee flexion eliminates the restraining effect of the M. gastrocnemius on ankle dorsiflexion and presents a video description of a standardized examination protocol.

诊断<em&gt;家鼠腓肠肌</em&gt;密封性 - 关键因素临床考试

Common foot and ankle pathologies have been linked to isolated Musculus gastrocnemius tightness (MGT). Various examination techniques have been described ...

Hydra, a Computer-Based Platform for Aiding Clinicians in Cardiovascular Analysis and Diagnosis

Cardiovascular diseases (CVDs) are the leading cause of death throughout the world. The total risk of developing CVD is determined by the combined effect of different cardiovascular risk factors (e.g., diabetes, raised blood pressure, unhealthy diet, tobacco use, stress, etc.) that commonly coexist and act multiplicatively. Most CVDs can be prevented by an early identification of the highest risk factors and an appropriate treatment. The stratification of cardiovascular risk factors involves a wide range of parameters and tests that specialists use in their clinical practice. In addition to cardiovascular (CV) risk stratification, ambulatory blood pressure monitoring (ABPM) also provides relevant information for diagnostic and treatment purposes. This work presents a list of protocols based on the Hydra platform, a web-based system for clinical decision support which incorporates a set of functionalities and services that are required for complete cardiovascular analysis, risk assessment, early diagnosis, treatment and monitoring of patients over time. The program includes tools for inputting and managing comprehensive patient data, organized into different checkups to track the evolution over time. It also has a risk stratification tool to compute a CV risk factor based upon several risk stratification tables of reference. Additionally, the program includes a tool that incorporates ABPM analysis and allows the extraction of valuable information by monitoring blood pressure over a specific period of time. Finally, the reporting service summarizes the most relevant information in a set of reports that aid clinicians in their clinical decision-making process.

This article presents a protocol based on Hydra &#8212; a web-based system for clinical decision support that integrates a full and detailed set of functionalities and services required by physicians for complete cardiovascular analysis, risk assessment, early diagnosis, treatment, and monitoring over time.

一种计算机辅助心血管分析诊断平台的研究

Cardiovascular diseases (CVDs) are the leading cause of death throughout the world. The total risk of developing CVD is determined by the combined effect ...

International Expert Consensus and Recommendations for Neonatal Pneumothorax Ultrasound Diagnosis and Ultrasound-guided Thoracentesis Procedure

Pneumothorax (PTX) represents accumulation of the air in the pleural space. A large or tension pneumothorax can collapse the lung and cause hemodynamic compromise, a life-threatening disorder. Traditionally, neonatal pneumothorax diagnosis has been based on clinical images, auscultation, transillumination, and chest X-ray findings. This approach may potentially lead to a delay in both diagnosis and treatment. The use of lung US in diagnosis of PTX together with US-guided thoracentesis results in earlier and more precise management. The recommendations presented in this publication are aimed at improving the application of lung US in guiding neonatal PTX diagnosis and management.

Pneumothorax is a common emergency and critical disease in newborn infants that needs rapid, clear diagnosis and timely treatment. Diagnosis and treatment based on chest X-rays are associated with delayed management and radiation damage. Lung ultrasound (US) provides useful guidance for rapid, accurate diagnosis and the precise thoracentesis of pneumothorax.

国际专家共识与新生儿肺功能超声诊断与超声引导性胸腔手术建议

Pneumothorax (PTX) represents accumulation of the air in the pleural space. A large or tension pneumothorax can collapse the lung and cause hemodynamic ...

Diagnosis and Surgical Treatment of Human Brucellar Spondylodiscitis

Brucellar spondylodiscitis (BS) is the most prevalent and significant osteoarticular presentation of human Brucellosis, which is commonly manifested in pastoral communities. It is difficult to differentially diagnose and usually leads to irreversible neurologic deficits and spinal deformities. The initial diagnosis of BS is based on clinical findings and radiographic assessments, and the confirmed diagnosis should be established by the isolation of Brucella species from the blood and/or the standard tube agglutination test. Differential diagnosis of multifocal BS from either degenerative disc diseases or tuberculosis is especially highlighted. The surgical approach, either endoscopic or open, is demonstrated in detail, accompanied by radiographic evidence of structural compression or severe instability. Further, the crucial surgical steps, including single-stage transforaminal decompression, debridement, interbody fusion, and internal fixation, are explained. Moreover, perioperative care and postoperative rehabilitation are also addressed. Taken together, this clinical algorithm presents a practical guide that has yielded substantially satisfactory outcomes in the past decades, which can also be introduced for large-scale application to manage human BS, especially in endemic regions.

We describe a clinical algorithm, based on decades of front-line experience of diagnosis and surgical treatment of human Brucellar spondylodiscitis in the largest medical center of the China's Xinjiang Pastoral Area.

人类布鲁塞拉性皮诺皮诺皮炎的诊断和手术治疗

Brucellar spondylodiscitis (BS) is the most prevalent and significant osteoarticular presentation of human Brucellosis, which is commonly manifested in ...

Testing Acetylcholine Followed by Adenosine for Invasive Diagnosis of Coronary Vasomotor Disorders

More than 50% of patients with signs and symptoms of myocardial ischemia undergoing coronary angiography have unobstructed coronary arteries. Coronary vasomotor disorders (impaired vasodilatation and/or enhanced vasoconstriction/spasm) represent important functional causes for such a clinical presentation. Although impaired vasodilatation may be assessed with non-invasive techniques such as positron emission tomography or cardiac magnetic resonance imaging, there is currently no reliable non-invasive technique for the diagnosis of coronary spasm available. Thus, invasive diagnostic procedures (IDP) have been developed for the diagnosis of coronary vasomotor disorders including spasm testing as well as assessment of coronary vasodilatation. The identification of the underlying type of disorder (so called endotype) allows the initiation of targeted pharmacological treatments. Despite the fact that such an approach is recommended by the current European Society of Cardiology guidelines for the management of chronic coronary syndromes based on the CorMicA study, comparability of results as well as multicenter trials are currently hampered by major differences in institutional protocols for coronary functional testing. This article describes a comprehensive IDP protocol including intracoronary acetylcholine provocation testing for diagnosis of epicardial/microvascular spasm, followed by Doppler wire-based assessment of coronary flow reserve (CFR) and hyperemic microvascular resistance (HMR) in search of coronary vasodilatory impairment.

Coronary vasomotion disorders represent frequent functional causes of angina in patients with unobstructed coronaries. The underlying mechanism of angina (endotype) in these patients can be determined by a comprehensive invasive diagnostic procedure based on acetylcholine provocation testing followed by Doppler-derived assessment of the coronary flow reserve and microvascular resistance.

检测乙酰胆碱，然后使用腺苷进行冠状动脉血管舒缩疾病的侵入性诊断

More than 50% of patients with signs and symptoms of myocardial ischemia undergoing coronary angiography have unobstructed coronary arteries. Coronary ...

Measurement of Myocardial Lactate Production for Diagnosis of Coronary Microvascular Spasm

In about a quarter of patients with angina and non-obstructive coronary arteries, no epicardial spasm is noted on coronary arteriography during an angina attack. Since the pressure-rate product is almost identical at rest and the onset of attack in those patients, the decrease in coronary blood flow rather than increased myocardial oxygen consumption is likely to explain myocardial ischemia, indicating a substantial involvement with coronary microvascular spasm (MVS). Myocardial lactate production, which could be defined as a negative myocardial lactate extraction ratio (ratio of the coronary arterial-venous difference in lactate concentration to arterial concentration), is considered indicative of objective evidence to support the emerging myocardial ischemia. Thus, monitoring of the myocardial lactate production and the emergence of chest pain and ischemic electrocardiographic changes during acetylcholine (ACh) provocation testing is of significant value for detecting the entity of MVS. Practically, 1 min after incremental doses of ACh (20, 50, and 100 &#956;g) are administered into the left coronary artery (LCA), paired samples of 1 mL of blood are collected from the LCA ostium and coronary sinus for measurement of lactate concentration by a calibrated automatic lactate analyzer. Then, the development of MVS could be confirmed by negative myocardial lactate extraction ratio despite the absence of angiographically demonstrable epicardial coronary spasm or before its occurrence throughout ACh provocation testing. In conclusion, assessment of myocardial lactate production is essential and valuable for the diagnosis of MVS.

Myocardial lactate production (coronary arterial-venous difference in serum lactate level) during coronary spasm provocation testing is considered as a highly sensitive marker that reflects acetylcholine-induced myocardial ischemia due to microvascular spasm. This article presents the procedures to assess myocardial lactate production for the diagnosis of coronary microvascular spasm.

测量心肌乳酸生成用于诊断冠状动脉微血管痉挛

In about a quarter of patients with angina and non-obstructive coronary arteries, no epicardial spasm is noted on coronary arteriography during an angina ...

High-speed Video Microscopy Analysis for First-line Diagnosis of Primary Ciliary Dyskinesia

Primary ciliary dyskinesia (PCD) is a congenital disorder predominantly inherited in an autosomal recessive trait. The disorder causes disturbance in the motion of cilia, leading to severe impairment of mucociliary clearance (MCC). If undiagnosed or diagnosed too late, the condition leads to the development of bronchiectasis and serious damage to the lungs in later life. Most of the methods for diagnosing PCD are time-consuming and demand extensive economic resources to establish them. High-speed video microscopy analysis (HSVMA) is the only diagnostic tool to visualize and analyze living respiratory cells with beating cilia in vitro. It is fast, cost-effective, and, in experienced hands, very reliable as a diagnostic tool for PCD. In addition, classical diagnostic measures such as transmission electron microscopy (TEM) are not applicable for some mutations as morphological changes are absent. 
This paper describes the process of collecting respiratory epithelial cells, the further preparation of the specimen, and the process of HSVMA. We also describe how brushed cells can be successfully kept unharmed and beating by keeping them in a nourishing medium for storage and transport to the investigation site in cases where a clinic does not possess the equipment to perform HSVMA. Also shown are videos with pathologic beating patterns from patients with a mutation in the dynein arm heavy chain 11 gene (DNAH11), which cannot be diagnosed with TEM; the result of an inconclusive HSVMA due to infection of the upper airways, as well as an unsuccessful brushing with superimposition of red blood cells. With this article, we would like to encourage every unit dealing with pulmonology patients and rare lung diseases to perform HSVMA as part of their daily routine diagnostics for PCD or send the specimens over to a center specializing in performing HSVMA.

High-speed video microscopy analysis is a relatively easy-to-perform, fast, cost-effective, and, in experienced hands, a considerably reliable tool for first-line diagnostics of primary ciliary dyskinesia, which should be available in every center involved in diagnostics and the treatment of severe lung diseases.

高速视频显微镜分析在原发性睫状体运动障碍一线诊断中的应用

Primary ciliary dyskinesia (PCD) is a congenital disorder predominantly inherited in an autosomal recessive trait. The disorder causes disturbance in the ...