本研究由龙岩市科技计划项目资助（批准号2020LYF17029）。

本研究中描述的方案已由福建医科大学龙岩第一附属医院伦理委员会根据《赫尔辛基宣言》确立的伦理准则进行审查和批准。人类参与者的安全和福祉是我们最关心的问题，所有程序都旨在将潜在的风险和不适降至最低。所有收集的数据将被保密处理，并仅用于本研究的目的。受试者可以随时自由退出研究，他们参与或退出的决定不会影响他们与研究人员或机构的关系。
1. 一般信息
<ol><li>选择诊断为胃 LGIN 的患者。本回顾性分析纳入2020年11月至2022年6月在龙岩第一医院、邢台第三医院、甘肃省肿瘤医院经组织病理学检查诊断为胃LGIN并接受ESD治疗的120例患者。</li>
</ol>2. 纳入和排除标准
<ol><li>纳入标准<ol><li>包括通过组织病理学检查诊断为胃 LGIN 并在诊断后 3 个月内接受 ESD 治疗的患者。</li>
		<li>确保所有活检和手术标本均根据世界卫生组织（WHO）消化系统肿瘤病理诊断标准7进行诊断，并在手术前后由两名病理学家审查所有病理切片。</li>
		<li>完整的临床数据。</li>
	</ol></li>
	<li>排除标准<ol><li>排除最近接受过抗生素、质子泵抑制剂 （PPI）、铋制剂和抑酸剂治疗 幽门螺 杆菌 （HP） 感染的患者。</li>
		<li>排除通过组织病理学检查诊断为胃高级别上皮内瘤变 （HGIN）、胃癌或其他转移性肿瘤的患者。</li>
		<li>排除接受手术的患者。</li>
		<li>排除接受放疗或化疗的患者。</li>
		<li>排除临床数据不完整的患者。</li>
	</ol></li>
</ol>3. 研究方法
<ol><li>内窥镜检查和手术<ol><li>让所有通过组织病理学检查诊断为胃 LGIN 的患者接受常规白光内窥镜检查 （WLE）。<ol><li>在进行内窥镜检查之前，请确保患者进行一些准备工作，包括禁食 6 小时以上。 注意:检查通常涉及喉咙局部麻醉，以尽量减少不适。</li>
			<li>将内窥镜插入患者的口腔，并逐渐通过食道将其推进到胃中。在WLE下观察消化道内部，包括粘膜中血管的颜色、形状、质地和分布，寻找可能的病变。</li>
			<li>根据观察结果采集组织样本或进行其他治疗操作。检查完成后，逐渐撤出内窥镜并结束检查。</li>
		</ol></li>
		<li>使用ME-NBI技术观察胃黏膜表面的微观结构，以进一步确定病变的程度。 注意:ME-NBI是一种形成模式，在WLE模式下观察近似粘膜后，通过内窥镜上的按钮进行切换。它可以观察传统内窥镜检查无法观察到的微血管 （MV） 和微观结构 （MS）。<ol><li>在 WLE 中发现可疑病灶后，从远处到近处，从正常到病灶中心放大。</li>
			<li>通过ME-NBI检查，判断是否存在分界线（DL）、MV和MS各向异性、病灶表面的一些特殊征象，更好地观察胃肠道黏膜表面的微观结构，进而判断病灶的性质，可能是病灶。</li>
		</ol></li>
		<li>接下来，用一次性粘膜切刀在患者病变边界外 3-5 毫米处做一个电凝标记，标记点之间的距离约为 2 毫米。</li>
		<li>通过特殊的内窥镜注射针将制备的注射液（250mL生理盐水+3mg肾上腺素+2mL靛蓝）注射到粘膜下层。 注意:生理盐水主要用于保持粘膜下层湿润，并将粘膜层与肌肉层分开。肾上腺素收缩血管，减少出血。靛蓝胭脂红是一种染料，可帮助医生更好地可视化粘膜下层血管的分布。分配的成分包括 250 mL 生理盐水 + 3 mg 肾上腺素 + 2 mL 靛蓝胭脂。</li>
		<li>病灶完全抬高后，用一次性粘膜切刀在距病灶标记点约 3 mm 处做一个环形切口，然后进行黏膜下夹层，直到病灶完全切除。</li>
		<li>解剖后，仔细观察伤口并使用电凝钳进行电凝。如有必要，使用金属止血夹夹住伤口。</li>
	</ol></li>
	<li>病理检查<ol><li>完全切除后，使用抽吸或圈套取出组织样本。<ol><li>固定并保存取出的组织样本，并在口腔和肛门两侧标记以进行病理检查和诊断。将样品在20-25°C下用10%福尔马林固定24-48小时。 注:病理描述包括病变的一般形态、体积和大小、边缘、组织学分类、浸润深度和胃粘膜病变的信息。</li>
		</ol></li>
	</ol></li>
	<li>HP检测方法<ol><li>使用快速脲酶试验或 13C 呼气试验。<ol><li>在快速脲酶试验中，在内窥镜检查下用活检钳取患者胃壁样本，并将其放入含有脲酶的试剂中。</li>
			<li>检查胃壁中是否存在 幽门螺杆 菌。脲酶分解尿素产生氨，使试剂呈碱性并变红。观察试剂颜色的变化并确定是否存在 幽门螺杆 菌感染。 注意:13C 呼气测试是一种非侵入性、无痛且无副作用的测试，通过口服含有 13C 标记尿素的尿素并使用同位素比质谱仪检测患者呼出的呼吸中 13C 标记的二氧化碳的含量。如果胃中存在 幽门 螺杆菌，细菌会分解13C标记的尿素产生13C标记的二氧化碳，通过测量呼出气体中13C标记的二氧化碳的量，可以确定是否存在 幽门螺 杆菌感染。需要注意的是，在进行快速脲酶试验或 13C 呼气试验之前，患者必须停止质子泵抑制剂 （PPI） 至少 2 周，并停止抗菌药物、铋和某些具有抗菌特性的草药至少 4 周。这些药物会影响检测结果的准确性。在13C呼气试验期间，患者还必须禁食或避免进食至少2小时，以避免食物对检测结果的影响。</li>
		</ol></li>
	</ol></li>
	<li>数据收集和分组<ol><li>汇总患者的一般临床资料，包括性别、年龄、 幽门螺杆菌 （HP）感染、病灶大小、病灶位置、形态、胃黏膜充血、结节状态、表面溃疡和糜烂、ME观察。</li>
		<li>根据所进行的评估，根据ESD后病理诊断是否升级，将研究中的所有患者分为升级组和非升级组。 注:升级组术后病理诊断为胃HGIN、早期胃癌（EGC）或晚期胃癌，而未升级组术后病理诊断为胃LGIN或炎症。</li>
	</ol></li>
</ol>4. 统计方法
<ol><li>使用统计软件来组织和分析所有数据。以频率（百分比）统计方式分析分类数据，并使用卡方检验或费舍尔精确检验进行比较。 注:本研究使用了 SPSS 软件。</li>
	<li>采用逻辑回归分析来识别诊断为胃 LGIN 的患者与 ESD 后病理升级相关的危险因素。小于 0.05 的 P 值表示存在统计显著差异。</li>
</ol>

胃低级别上皮内瘤变风险预测模型

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作者声明他们没有利益冲突。

胃LGIN是一种常见的胃肠道疾病，随着患者人数的不断增加，该病的预防和治疗变得越来越重要。建立风险预测模型对于指导临床治疗和预防疾病发生至关重要8.胃LGIN的病理升级是指LGIN性质在一定时期内恶化，这可能导致LGIN转化为HGIN甚至更严重的情况。在临床实践中，病理升级的预测已成为医务人员关注的焦点。目前，已经研究了一些基于临床表现、影像学表现和生物标志物的预测模型，但这些模型存在指标复杂、长期消耗的缺点。因此，有必要开发一个简单有效的预测模型。根据目前国内共识建议，应积极采用ESD治疗具有病理升级高危因素的胃LGIN患者。然而，越来越多的研究表明，胃 LGIN 患者的活检病理结果与术后病理结果之间存在显着差异9。 
研究显示，在观察的120例胃LGIN患者中，有49例在术后进行了病理升级，病理升级率为40.83%。与其他相关研究相比，这一比率相对较高。这些发现的可能原因包括低估了病理活检的结果。内窥镜医师资格的差异可能导致患者病情和活检部位评估的差异，导致表面或不充分的取样和部位偏差。结果的准确性也可能受到病理学家的认知能力和知识的影响。此外，活检的准确性可能会受到不同内窥镜技术的影响。例如，应用高清胃镜观察病变的微血管和微观结构形态，确定靶向活检最显眼的部位，可以大大提高活检的精确度。
本研究表明，胃LGIN患者年龄越大，术后病理升级的风险越高，这与国内其他研究结果相似10。因此，对于60岁以上的胃LGIN患者，应进行密切随访，如果患者有更多危险因素，建议及时手术。关于病灶大小，本研究表明，病灶大小≥2 cm是病理升级的独立危险因素，与其他研究结果基本一致11。胃LGIN患者手术治疗后可能影响病理升级发生的病灶大小临界值仍存在争议，需要临床研究寻找最合适的临界值。应使用大样本量方法进行进一步研究。结果显示，黏膜表面充血、溃疡、糜烂是胃LGIN患者术后病理升级的独立危险因素，与相关研究相似12。病变表面溃疡和糜烂更容易引起手术后病理升级，这可能是因为胃黏膜反复损伤会促进异型增生和肠上皮化生，从而增加胃癌的风险。 
然而，这项研究遇到了一定的局限性。首先，作为一项回顾性研究，无法完全控制偏倚和其他混杂因素。今后应开展前瞻性研究，以验证结论，增强客观性和推理能力。其次，由于样本数量有限，无法将样本分离到验证集中，以便对模型进行外部验证。因此，必须从多个中心收集额外的患者数据，以增加样本数量并扩大观察到的变量范围。这将提高模型的预测效果和结果的可信度。此外，进行外部验证将增强模型的可靠性。 
综上所述，本研究通过分析接受手术的胃LGIN患者病理升级的独立危险因素而生成的风险预测模型具有显著的预测价值，可为临床环境中的LGIN治疗提供有益的指导。

胃癌是最常见的恶性肿瘤之一，尤其是在东亚地区，发病率和死亡率都很高。它是中国最常见的癌症之一，新诊断和死亡约占全球总数的一半 1.它是中国人群发病和死亡的主要原因2.胃癌在肿瘤中死亡率最高，居世界第三位，其预后高度依赖于病变的分期1。晚期患者的 5 年生存率低于 30%，而早期患者的 5 年生存率通常在 90% 以上。因此，胃癌的早期诊断和治疗对于预防和控制疾病至关重要3. 
Correa级联反应被广泛认为是胃癌发展的主要模式之一，表明胃癌的致癌过程逐渐从萎缩性胃炎发展为肠上皮化生、上皮内瘤变，最后发展为腺癌4。随着现代技术的应用和胃镜检查的日益普及，越来越多的胃黏膜癌前病变被检测到，包括放大内窥镜 （ME）、染色内窥镜和窄带成像 （NBI）5。 
低级别上皮内瘤变（LGIN）是胃癌的癌前病变之一，与胃癌密切相关。然而，与活检结果相比，一些低级别上皮内瘤变患者在内镜黏膜下夹层 （ESD） 后表现出病理升级6。因此，在临床实践中，对于活检证实的LGIN患者的随访或治疗选择存在一些争议。本文探讨了胃LGIN患者ESD治疗后病理升级的危险因素，建立并评价了LGIN发生的风险预测模型，为胃LGIN患者的临床管理提供了更科学、更有价值的参考意见。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Disposable mucosal incision knife</td>
			<td>Olympus (Japan)</td>
			<td>KD-650Q</td>
			<td></td>
		</tr>
		<tr>
			<td>Endoscopic image processing device</td>
			<td>Olympus (Japan)</td>
			<td>CV-290</td>
			<td></td>
		</tr>
		<tr>
			<td>Hemostasis Clips</td>
			<td>MICRO-TECH(Nanjing)</td>
			<td>ROCC-D-26-195</td>
			<td></td>
		</tr>
		<tr>
			<td>High-frequency hemostatic forceps</td>
			<td>Olympus (Japan)</td>
			<td>FD-410LR</td>
			<td></td>
		</tr>
		<tr>
			<td>Indicarminum</td>
			<td>MICRO-TECH(Nanjing)</td>
			<td>MTN-DYZ-15</td>
			<td></td>
		</tr>
		<tr>
			<td>Injection Needles</td>
			<td>MICRO-TECH(Nanjing)&#160;</td>
			<td>IN02-25423230</td>
			<td></td>
		</tr>
		<tr>
			<td>Magnifying gastroscope</td>
			<td>Olympus (Japan)</td>
			<td>GIF-H290Z</td>
			<td></td>
		</tr>
		<tr>
			<td>Orthodontic rubber band</td>
			<td>3M Unitek Corporation</td>
			<td></td>
			<td>6.4 mm 3.5 oz</td>
		</tr>
		<tr>
			<td>Therapeutic gastroscopy</td>
			<td>Olympus (Japan)</td>
			<td>GIF-2TQ260M</td>
			<td></td>
		</tr>
		<tr>
			<td>Transparent cap</td>
			<td>Olympus (Japan)</td>
			<td>D-201-11804</td>
			<td></td>
		</tr>
	</tbody>
</table>

establishment and evaluation of a risk prediction model for pathological escalation of gastric low-grade intraepithelial neoplasia

本研究旨在探讨胃低级别上皮内皮瘤变（LGIN）内镜手术后病理升级的危险因素，并建立和评估LGIN的风险预测模型。回顾性分析了 2020 年 11 月至 2022 年 6 月期间通过活检和内窥镜粘膜下剥离术 （ESD） 诊断为胃 LGIN 的 120 例患者。收集所有患者的性别、 年龄、 HP感染、病灶大小、病灶位置、形态、胃黏膜充血、结节状态、表面溃疡和糜烂、ME观察，根据活检和ESD术后病理诊断结果分为升级组和非升级组。采用logistic回归分析筛选ESD手术治疗后病理升级的独立危险因素，建立风险预测模型。120例胃LGIN患者中，49例出现术后病理升级;病理升级率为40.83%。其中，42例患者升级为高级别上皮内瘤变（HGIN），1例升级为晚期胃癌，6例升级为早期胃癌（EGC）。单因素分析显示，两组患者年龄、病灶大小、胃黏膜充血、表面溃疡、糜烂等差异有统计学意义（p < 0.05）。多因素Logistic回归分析显示，年龄≥60岁、焦距≥2 cm、胃黏膜淤血、表面溃疡和糜烂是胃LGIN患者术后病理恶化的独立危险因素。P = 1/[1 + e（26.515-0.161 x β1-0.357 x β2+0.039 x β3-0.269 x β4）] 的最终联合概率预测模型。年龄、病灶大小≥2 cm、胃黏膜充血、病灶表面溃疡和糜烂是胃LGIN患者术后病理升级的危险因素。本研究建立的基于危险因素的风险预测模型具有预测价值，可为胃LGIN患者的临床治疗提供科学参考。

ESD后胃LGIN患者病理升级的发生率 本研究共纳入120例胃LGIN患者，其中49例（40.83%）在ESD后出现病理升级。其中，42例升级为HGIN，1例升级为晚期胃癌，6例升级为EGC。共有71例未经历病理升级，其中2例降级为炎症，69例仍为LGIN，病理降级率为1.67%。
胃LGIN患者ESD后病理升级的单因素分析 如 表1所示，胃LGIN患者术后病理升级组与非升级组在性别、HP感染、病灶位置、表面结节、ME层分界线（DL）、病灶形态、微观结构（MS）和微血管形态方面差异无统计学意义（p > 0.05）。然而，在年龄、病灶大小、胃黏膜充血以及表面溃疡和糜烂方面存在统计学差异（p < 0.05）。
胃LGIN患者ESD后病理升级的多因素logistic回归分析 单因素分析筛选出p<0.05的因素，包括年龄、病灶大小、胃黏膜充血、表面溃疡和糜烂等因素，作为自变量，胃LGIN患者ESD后病理升级作为logistic回归分析的因变量（表2）。 表3 显示，年龄≥60岁、病灶直径≥2 cm、胃黏膜充血、表面溃疡和糜烂是胃LGIN患者ESD后病理升级的独立危险因素。基于逻辑回归的预测公式，最终的联合概率预测模型为 P = 1 / [1 + e（26.515-0.161xβ1-0.357xβ2+0.039xβ3-0.269xβ4）]:P（y = 1/x） = 1/（1+e-（β0+β1x1+β2x2+β3x3+β4x4）），其中 y 表示二分因变量，x 表示自变量向量，β 表示回归系数，e 表示自然对数的底数。此外，技术术语缩写将在首次使用时定义。该公式表明，给定自变量 x，事件发生 y = 1 的概率为 P（y = 1/x）。当 β0 + β1 x 1 + β2 x 2 + β3 x 3 + β4 x 4 的总和增加时，P（y = 1/x） 的值也增加，表明事件发生 y = 1 的概率更高;相反，事件发生 y = 1 的概率较低。其中，β1为年龄回归系数，β2为病灶大小回归系数，β3为胃黏膜淤血回归系数，β4为表面溃疡糜烂回归系数。
案例介绍 代表性病例为一名50岁女性患者，体格检查时出现黏膜粗糙、胃角糜烂。内镜活检证实低级别上皮内瘤变 （LGIN）。使用放大内窥镜检查、窄带成像 （NBI） 和色素内窥镜检查的进一步研究表明，患者有发生高级别上皮内瘤变 （HGIN） 的风险。建议患者接受内镜黏膜下剥离（ESD）治疗，最终病理诊断为HGIN（ 如图1所示）。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/65868/65868fig01.jpg"> 图1:案例介绍。 （A、B） 术前，WLE 显示胃黏膜粗糙伴糜烂。（C，D）在ME + NBI观察到的病变中观察到不规则的MV和MS。（E）靛蓝染色可以看到病变的轮廓。（F）在内窥镜下标记病灶。（G） 粘膜下水注射后剥离。（八）伤口止血治疗。（一）将固定标本送去进行病理检查。<a href="https://www.jove.com/files/ftp_upload/65868/65868fig01large.jpg" target="_blank">请点击这里查看此图的较大版本.</a>
表1:ESD后病理升级的单因素分析（%）。<a href="https://www.jove.com/files/ftp_upload/65868/Table1.xlsx" target="_blank">请按此下载此表格。</a>
表2:ESD后病理升级的多变量logistic回归分析变量分配表。<a href="https://www.jove.com/files/ftp_upload/65868/Table%202.xlsx" target="_blank">请按此下载此表格。</a>
表3:ESD后病理升级的多因素logistic回归分析。<a href="https://www.jove.com/files/ftp_upload/65868/Table%203.xlsx" target="_blank">请按此下载此表格。</a>

Watch this Scientific Journal Video about Establishment and Evaluation of a Risk Prediction Model for Pathological Escalation of Gastric Low-Grade Intraepithelial Neoplasia at JoVE.com

作者聚焦:推进胃肠道肿瘤的早期检测和治疗

在这里，我们对既往内镜活检诊断为胃低级别上皮内瘤变的患者进行了系统评估，并通过内镜粘膜下剥离术 （ESD） 完全切除病灶获得病理诊断，分析可能增加病理升级风险的因素。

胃低级别上皮内瘤变病理升级风险预测模型的建立与评价

The study aims to explore the risk factors for pathological escalation after endoscopic surgery for gastric low-grade intraepithelial neoplasia (LGIN) and ...

我们的研究范围是早期胃肠道肿瘤的检测、诊断和治疗。我想回答的问题是如何预测胃核病理性升级为上皮核的风险，以便对患者进行诊断和首次治疗。消化内窥镜检查的最新发展涉及微创技术、改进成像方式、翻修活检和机器人辅助，这当然可以改善诊断和可怕的结果。未来，我们将重点研究如何提高早期胃癌的诊断率，包括通过人工智能和先进的内窥镜技术对早期胃癌的诊断设想。

establishment-evaluation-risk-prediction-model-for-pathological

Research

JoVE Journal

Cancer Research

Establishment and Evaluation of a Risk Prediction Model for Pathological Escalation of Gastric Low-Grade Intraepithelial Neoplasia

923 次观看.  Longyan First Affiliated Hospital of Fujian Medical University. 我们的研究范围是早期胃肠道肿瘤的检测、诊断和治疗。我想回答的问题是如何预测胃核病理性升级为上皮核的风险，以便对患者进行诊断和首次治疗。消化内窥镜检查的最新发展涉及微创技术、改进成像方式、翻修活检和机器人辅助，这当然可以改善诊断和可怕的结果。未来，我们将重点研究如何提高早期胃癌的诊断率，包括通过人工智能和先进的内窥镜技术对早期?...

视频: 作者聚焦:推进胃肠道肿瘤的早期检测和治疗

Characterization of Cell Membrane Extensions and Studying Their Roles in Cancer Cell Adhesion Dynamics

The cell membrane&#39;s extension repertoire modulates various malignant behaviors of cancer cells, including their adhesive and migratory potentials. The ability to accurately classify and quantify cell extensions and measure the effect on a cell&#39;s adhesive capacity is critical to determining how cell-signaling events impact cancer cell behavior and aggressiveness. Here, we describe the in vitro design and use of a cell extension quantification method in conjunction with an adhesion capacity assay in an established in vitro model for adrenocortical carcinoma (ACC). Specifically, we test the effects of DKK3, a putative tumor suppressor and a pro-differentiation factor, on the membrane extension phenotype of the ACC cell line, SW-13. We propose these assays to provide relatively simple, reliable, and easily interpretable metrics to measures these characteristics under various experimental conditions.

This study demonstrates the utility and ease of quantitative cell membrane extension measurement and its correlation to adhesive capacity of cells. As a representative example, we show here that Dickkopf-related protein 3 (DKK3) promotes increased lobopodia formation and cell adhesiveness in adrenocortical carcinoma cells in vitro.

细胞膜延伸特性及其在癌细胞黏附动力学中的作用研究

The cell membrane&#39;s extension repertoire modulates various malignant behaviors of cancer cells, including their adhesive and migratory potentials. The ...

科研

癌症研究

Establishment of a Primary Culture of Patient-derived Soft Tissue Sarcoma

Soft tissue sarcomas (STS) represent a spectrum of heterogeneous malignancies with a difficult diagnosis, classification, and management. To date, more than 50 histological subtypes of these rare solid tumors have been identified. Thus, due to their extraordinary diversity and low incidence, our understanding of the biology of these tumors is still limited. Patient-derived cultures represent the ideal platform to study STS pathophysiology and pharmacology. We thus developed a human preclinical model of STS starting from tumor specimens harvested from patients undergoing surgical resection. Patient-derived STS cell cultures were obtained from the surgical specimens by collagenase digestion and isolated by filtration. Cells were counted, seeded, and left for 14 days in standard monolayer cultures and then processed by downstream analysis. Before performing molecular or pharmaceutical analyses, the establishment of STS primary cultures was confirmed through the evaluation of cytomorphologic features and, when available, immunohistochemical markers. This method represents a useful tool 1) to study the natural history of these poorly explored malignancies and 2) to test the effects of different drugs in an effort to learn more about their mechanisms of action.

The following protocol focuses on the establishment of a primary culture of patient-derived soft tissue sarcoma (STS). This model could help us to better understand the molecular background and pharmacological profile of these rare malignancies and could represent a starting point for further research aimed at improving STS management.

病人衍生软组织肉瘤原代培养的建立

Soft tissue sarcomas (STS) represent a spectrum of heterogeneous malignancies with a difficult diagnosis, classification, and management. To date, more ...

Utilization of Ultrasound Guided Tissue-directed Cellular Implantation for the Establishment of Biologically Relevant Metastatic Tumor Xenografts

Preclinical testing of anticancer therapies relies on relevant xenograft models that mimic the innate tendencies of cancer. Advantages of standard subcutaneous flank models include procedural ease and the ability to monitor tumor progression and response without invasive imaging. Such models are often inconsistent in translational clinical trials and have limited biologically relevant characteristics with low proclivity to produce metastasis, as there is a lack of a native microenvironment. In comparison, orthotopic xenograft models at native tumor sites have been shown to mimic the tumor microenvironment and replicate important disease characteristics such as distant metastatic spread. These models often require tedious surgical procedures with prolonged anesthetic time and recovery periods. To address this, cancer researchers have recently utilized ultrasound-guided injection techniques to establish cancer xenograft models for preclinical experiments, which allows for rapid and reliable establishment of tissue-directed murine models. Ultrasound visualization also provides a noninvasive method for longitudinal assessment of tumor engraftment and growth. Here, we describe the method for ultrasound-guided injection of cancer cells, utilizing the adrenal gland for NB and renal sub capsule for ES. This minimally invasive approach overcomes tedious open surgery implantation of cancer cells in tissue-specific locations for growth and metastasis, and abates morbid recovery periods. We describe the utilization of both established cell lines and patient derived cell lines for orthotopic injection. Pre-made commercial kits are available for tumor dissociation and luciferase tagging of cells. Injection of cell suspension using image-guidance provides a minimally invasive and reproducible platform for the creation of preclinical models. This method is utilized to create reliable preclinical models for other cancers such as bladder, liver and pancreas exemplifying its untapped potential for numerous cancer models.

Here, we present a protocol to utilize ultrasound-guided injection of neuroblastoma (NB) and Ewing's sarcoma (ES) cells (established cell lines and patient-derived tumor cells) at biologically relevant sites to create reliable preclinical models for cancer research.

超声引导组织定向细胞植入术在生物学相关转移性肿瘤移植中的应用

Preclinical testing of anticancer therapies relies on relevant xenograft models that mimic the innate tendencies of cancer. Advantages of standard ...

Morphology-Based Distinction Between Healthy and Pathological Cells Utilizing Fourier Transforms and Self-Organizing Maps

The appearance and the movements of immune cells are driven by their environment. As a reaction to a pathogen invasion, the immune cells are recruited to the site of inflammation and are activated to prevent a further spreading of the invasion. This is also reflected by changes in the behavior and the morphological appearance of the immune cells. In cancerous tissue, similar morphokinetic changes have been observed in the behavior of microglial cells: intra-tumoral microglia have less complex 3-dimensional shapes, having less-branched cellular processes, and move more rapidly than those in healthy tissue. The examination of such morphokinetic properties requires complex 3D microscopy techniques, which can be extremely challenging when executed longitudinally. Therefore, the recording of a static 3D shape of a cell is much simpler, because this does not require intravital measurements and can be performed on excised tissue as well. However, it is essential to possess analysis tools that allow the fast and precise description of the 3D shapes and allows the diagnostic classification of healthy and pathogenic tissue samples based solely on static, shape-related information. Here, we present a toolkit that analyzes the discrete Fourier components of the outline of a set of 2D projections of the 3D cell surfaces via Self-Organizing Maps. The application of artificial intelligence methods allows our framework to learn about various cell shapes as it is applied to more and more tissue samples, whilst the workflow remains simple.

Here, we provide a workflow that allows the identification of healthy and pathological cells based on their 3-dimensional shape. We describe the process of using 2D projection outlines based on the 3D surfaces to train a Self-Organizing Map that will provide objective clustering of the investigated cell populations.

利用傅里叶变换和自组织映射的形态学区分健康和病理细胞

The appearance and the movements of immune cells are driven by their environment. As a reaction to a pathogen invasion, the immune cells are recruited to ...

Evaluation of Colorectal Cancer Risk and Prevalence by Stool DNA Integrity Detection

Nowadays, stool DNA can be isolated and analyzed by several methods. The long fragments of DNA in stool can be detected by a qPCR assay, which provides a reliable probability of the presence of pre-neoplastic or neoplastic colorectal lesions. This method, called fluorescence long DNA (FL-DNA), is a fast, non-invasive procedure that is an improvement upon the primary prevention system. This method is based on evaluation of fecal DNA integrity by quantitative amplification of specific targets of genomic DNA. In particular, the evaluation of DNA fragments longer than 200 bp allows for detection of patients with colorectal lesions with very high specificity. However, this system and all currently available stool DNA tests present some general issues that need to be addressed (e.g., the frequency at which tests should be carried out and optimal number of stool samples collected at each timepoint for each individual). However, the main advantage of FL-DNA is the possibility to use it in association with a test currently used in the CRC screening program, known as the immunochemical-based fecal occult blood test (iFOBT). Indeed, both tests can be performed on the same sample, reducing costs and achieving a better prediction of the eventual presence of colorectal lesions.

The presented diagnostic FL-DNA kit is a time-saving and user-friendly method to determine the reliable probability of the presence of colorectal cancer lesions.

通过大刀DNA完整性检测评估结肠直肠癌风险和患病率

Nowadays, stool DNA can be isolated and analyzed by several methods. The long fragments of DNA in stool can be detected by a qPCR assay, which provides a ...

Establishment of Gastric Cancer Patient-derived Xenograft Models and Primary Cell Lines

The use of preclinical models to advance our understanding of tumor biology and investigate the efficacy of therapeutic agents is key to cancer research. Although there are many established gastric cancer cell lines and many conventional transgenic mouse models for preclinical research, the disadvantages of these in vitro and in vivo models limit their applications. Because the characteristics of these models have changed in culture, they no longer model tumor heterogeneity, and their responses have not been able to predict responses in humans. Thus, alternative models that better represent tumor heterogeneity are being developed. Patient-derived xenograft (PDX) models preserve the histologic appearance of cancer cells, retain intratumoral heterogeneity, and better reflect the relevant human components of the tumor microenvironment. However, it usually takes 4-8 months to develop a PDX model, which is longer than the expected survival of many gastric patients. For this reason, establishing primary cancer cell lines may be an effective complementary method for drug response studies. The current protocol describes methods to establish PDX models and primary cancer cell lines from surgical gastric cancer samples. These methods provide a useful tool for drug development and cancer biology research.

The current protocol describes methods to establish patient-derived xenograft (PDX) models and primary cancer cell lines from surgical gastric cancer samples. The methods provide a useful tool for drug development and cancer biology research.

建立胃癌患者衍生异种移植模型和原细胞系

The use of preclinical models to advance our understanding of tumor biology and investigate the efficacy of therapeutic agents is key to cancer research. ...

Establishing a Competing Risk Regression Nomogram Model for Survival Data

The Kaplan&#8211;Meier method and Cox proportional hazards regression model are the most common analyses in the survival framework. These are relatively easy to apply and interpret and can be depicted visually. However, when competing events (e.g., cardiovascular and cerebrovascular accidents, treatment-related deaths, traffic accidents) are present, the standard survival methods should be applied with caution, and real-world data cannot be correctly interpreted. It may be desirable to distinguish different kinds of events that may lead to the failure and treat them differently in the analysis. Here, the methods focus on using the competing regression model to identify significant prognostic factors or risk factors when competing events are present. Additionally, nomograms based on a proportional hazard regression model and a competing regression model are established to help clinicians make individual assessments and risk stratifications in order to explain the impact of controversial factors on prognosis.

Presented here is a protocol to build nomograms based on the Cox proportional hazards regression model and competing risk regression model. The competing method is a more rational method to apply when competing events are present in the survival analysis.

为生存数据建立竞争风险回归 Nomogram 模型

The Kaplan&#8211;Meier method and Cox proportional hazards regression model are the most common analyses in the survival framework. These are relatively ...

Pathological Analysis of Lung Metastasis Following Lateral Tail-Vein Injection of Tumor Cells

Metastasis, the primary cause of morbidity and mortality for most cancer patients, can be challenging to model preclinically in mice. Few spontaneous metastasis models are available. Thus, the experimental metastasis model involving tail-vein injection of suitable cell lines is a mainstay of metastasis research. When cancer cells are injected into the lateral tail-vein, the lung is their preferred site of colonization. A potential limitation of this technique is the accurate quantification of the metastatic lung tumor burden. While some investigators count macrometastases of a pre-defined size and/or include micrometastases following sectioning of tissue, others determine the area of metastatic lesions relative to normal tissue area. Both of these quantification methods can be exceedingly difficult when the metastatic burden is high. Herein, we demonstrate an intravenous injection model of lung metastasis followed by an advanced method for quantifying metastatic tumor burden using image analysis software. This process allows for investigation of multiple end-point parameters, including average metastasis size, total number of metastases, and total metastasis area, to provide a comprehensive analysis. Furthermore, this method has been reviewed by a veterinary pathologist board-certified by the American College of Veterinary Pathologists (SEK) to ensure accuracy.

Intravenous injection of cancer cells is often used in metastasis research, but the metastatic tumor burden can be difficult to analyze. Herein, we demonstrate a tail-vein injection model of metastasis and include a novel approach to analyze the resulting metastatic lung tumor burden.

肿瘤细胞外侧尾静脉注射后肺转移的病理分析

Metastasis, the primary cause of morbidity and mortality for most cancer patients, can be challenging to model preclinically in mice. Few spontaneous ...

Characterization and Functional Prediction of Bacteria in Ovarian Tissues

The theory of a "sterile" female upper reproductive tract has been encountering increasing opposition due to advancements in bacterial detection. However, whether ovaries contain bacteria has not yet been confirmed yet. Herein, an experiment to detect bacteria in ovarian tissues was introduced. We chose ovarian cancer patients in the cancer group and noncancerous patients in the control group. 16S rRNA gene sequencing was used to differentiate bacteria in ovarian tissues from the cancer and control groups. Furthermore, we predicted the functional composition of the identified bacteria by using BugBase and PICRUSt. This method can also be used in other viscera and tissues since many organs have been proven to harbor bacteria in recent years. The presence of bacteria in viscera and tissues may help scientists evaluate cancerous and normal tissues and may be aid in the treatment of cancer.

Immunohistochemistry staining and 16S ribosomal RNA gene (16S rRNA gene) sequencing were performed in order to discover and distinguish bacteria in cancerous and noncancerous ovarian tissues in situ. The compositional and functional differences of the bacteria were predicted by using BugBase and Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt).

卵巢组织细菌的特征和功能预测

The theory of a "sterile" female upper reproductive tract has been encountering increasing opposition due to advancements in bacterial detection. However, ...

An R-Based Landscape Validation of a Competing Risk Model

The Cox proportional hazard model is widely applied for survival analyses in clinical settings, but it is not able to cope with multiple survival outcomes. Different from the traditional Cox proportional hazard model, competing risk models consider the presence of competing events and their combination with a nomogram, a graphical calculating device, which is a useful tool for clinicians to conduct a precise prognostic prediction. In this study, we report a method for establishing the competing risk nomogram, that is, the evaluation of its discrimination (i.e., concordance index and area under the curve) and calibration (i.e., calibration curves) abilities, as well as the net benefit (i.e., decision curve analysis). In addition, internal validation using bootstrap resamples of the original dataset and external validation using an external dataset of the established competing risk nomogram were also performed to demonstrate its extrapolation ability. The competing risk nomogram should serve as a useful tool for clinicians to predict prognosis with the consideration of competing risks.

The present protocol describes codes in R for evaluating the discrimination and calibration abilities of a competing risk model, as well as codes for the internal and external validation of it.