The authors have nothing to acknowledge.

所有的程序进行审查和忠南国立大学医院的机构审查委员会批准。 1.测量与超声<ol><li>将患者的体位。 <ol><li>奠定了病人的手臂仰卧位，用床上双臂。 </li></ol></li><li>确定在患者的上肢的测量部位。 <ol><li>标记在手腕折痕的中点，内侧和在肘部的水平横向上髁，以及由钢笔二头肌沟之间的中间点。 </li><li>使用笔直接被统治者这三点连接。标记上的上臂和前臂（ 图1）的两个目标区域。 <ol><li>马克近端部分的10cm近端沿着肘部和二头肌沟之间的界限的拐点。 </li><li>马克远端部分的10cm远端沿着肘部和腕部之间的界限的拐点。 </li></ol></li></ol></li><li>测量上肢的皮下组织和肌肉的厚度。该过程应该由熟练的超声检查进行。 <ol><li>切换超声系统。通过键盘输入的软组织的2D模式。深度为5厘米。 <ol><li>选择14L5线阵探头。 </li><li>施加足够的凝胶的超声换能器。专注于皮下组织。 </li></ol></li><li>放置超声换能器垂直于上肢腹轴。使用短轴观。捕获图像，其中凝胶的厚度至少为1厘米，软组织轮廓不失真。 </li><li>测量肌肉和皮下组织的厚度。 <ol><li>由通过键盘超声系统的选择在软件校准绘制一条线</li><li>测量肌肉的厚度，骨的边界上限定为距离从最高点到最高点在面板的边界部分（ 图2）。 </li><li>测量皮下组织的厚度定义为从皮肤到筋膜的距离。 </li></ol></li></ol></li></ol> 2.超声临床应用BCRL <ol><li>测量在基线上肢的皮下组织和肌肉的厚度。 <ol><li>在步骤1中所示的顺序执行基线测量。 </li></ol></li><li>指导患者进行一系列使用0.5千克的哑铃进行14公屋的。 <ol><li>给病人压力袜或绷带多层穿。选择0.5千克的哑铃进行。 </li><li>指导患者在以下顺序执行的PRE:（1）哑铃飞，（2）三头肌伸展，（3）一个臂俯身行，（4）二头肌卷曲，（5）哑铃侧培养，和（6 ）前起重臂（ 图3）。 <ol><li>指导患者完成6练习，一天两次，按照以下时间表:在第1 次周各5次，在第 2周各10次，在第 3 次每周每15次， 第 4周期间每20次，25次每5 次 -8 个星期（ 图4）中。 </li></ol></li></ol></li><li>使用超声来确定相对于传统疗法的PRE的治疗效果。 <ol><li>测量皮下组织和上肢肌肉的厚度在4和8周。在步骤1中所示的顺序进行测量。 </li></ol></li><li>计算皮下组织和肌肉在基线，第4周，8周的厚度。 </li><li>分析数据。 <ol><li>比较使用配对t检验两个臂之间的差异。比较采用重复测量方差分析皮下组织和肌肉厚度的变化。 </li></ol></li></ol>

<ol>
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The authors have nothing to disclose.

BCRL的评估可以通过各种技术，包括臂圆周带测量，水位移测量，生物电阻抗谱，perometry（光电容积计），淋巴，临床医生的诊断，或肿胀由患者自我报告进行。然而，存在用于BCRL的测量没有标准;治疗的诊断和持续时间可根据经验在许多方式来确定，这取决于临床。 在临床上，用卷尺测量臂围是最流行，最方便的方法。然而，这种方法不能评价在皮下组织结构变化，以及错?...

淋巴水肿定义为局部富含蛋白质的液体潴留和组织肿胀折衷淋巴系统1,2引起的病症。如淋巴水肿的进行，纤维细胞和/或脂肪细胞增殖在受影响的地区，从而导致在皮肤和皮下组织和增加的脆弱性细菌和真菌感染3,4-质地变化。这些变化在四肢的各部分的观测可以进一步阐明的严重性和淋巴水肿的程度。该筋膜淋巴系统有助于淋巴水肿的严重程度超过epifascial淋巴系统。 最近的研究主要集中在5,6淋巴水肿的诊断使用超声检查。超声检查是一种相对廉价的观察软组织特性的方法。先前的研究报告说，淋巴水肿可以通过比较T为可靠的诊断与超声他厚度的皮肤和皮下组织，并且还通过在临床环境7,8-评估皮下组织的合规性。此外，超声被广泛应用于临床，因为其容易获得，从辐射照射的安全性，和廉价5,6。尽管有这些好处，在超声检查淋巴水肿的研究只集中在诊断，不治疗干预的评价。 因此，目前的研究旨在确定在阐明对BCRL PRE的影响不仅超的实用性，但也超声检查是否具有临床意义在确认疗效的后续测试。本研究旨在建立执行检查淋巴水肿超声波的协议。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>ACUSON S2000</td>
			<td>Siemens</td>
			<td>10041461</td>
			<td></td>
		</tr>
		<tr>
			<td>Eco Gel 99</td>
			<td>Seung Won Medical Corp.</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

ultrasonographic evaluation of breast cancer-related lymphedema

淋巴水肿是乳腺癌手术后最常见的并发症之一。有淋巴水肿的很多诊断工具，但没有标准方法还不存在。渐进性练习（PRE）有望改善淋巴水肿，无需额外肿胀。本研究通过使用超声来测量在肌肉和皮下组织的厚度变化显示对淋巴水肿的PRE的治疗效果。皮下组织的厚度比在非预组降低多在PRE基。超声检查被广泛应用于许多诊所因其交通方便，安全和廉价的。 超声波是诊断和乳腺癌相关淋巴水肿（BCRL）确定治疗效果的最佳工具之一。

这项研究包括32例确诊为BCRL。肌肉和皮下组织以及近端和远端上肢的圆周的厚度在基线，第4周，和PRE 8周后测得的。审查员测量上肢的周长用卷尺，然后将肌肉和皮下组织的厚度通过超声测量。患者被随机分为PRE组和非PRE基。提醒参加者不透露前测量和评估会议的小组作业。 在基线，通过超声测量的所有参与者的初始肌厚度?...

Watch this Scientific Journal Video about Ultrasonographic Evaluation of Breast Cancer-related Lymphedema at JoVE.com

Ultrasonographic Evaluation of Breast Cancer-related Lymphedema

这对诊断和随访试验研究利用超声检查对乳腺癌相关淋巴水肿（BCRL）确认渐进抗阻力训练（PRE）的治疗效果。超声能有效地应用于确保诊断和治疗淋巴水肿。

乳腺癌相关的淋巴水肿的超声评价

Lymphedema is one of the most common complications after breast cancer surgery. There are many diagnostic tools for lymphedema, but no standard method yet ...

ultrasonographic-evaluation-of-breast-cancer-related-lymphedema

Research

JoVE Journal

Cancer Research

9.1K 次观看.  Chungnam National University Hospital (CNUH). 这对诊断和随访试验研究利用超声检查对乳腺癌相关淋巴水肿（BCRL）确认渐进抗阻力训练（PRE）的治疗效果。超声能有效地应用于确保诊断和治疗淋巴水肿。 

视频: Ultrasonographic Evaluation of Breast Cancer-related Lymphedema

Utilizing Functional Genomics Screening to Identify Potentially Novel Drug Targets in Cancer Cell Spheroid Cultures

The identification of functional driver events in cancer is central to furthering our understanding of cancer biology and indispensable for the discovery of the next generation of novel drug targets. It is becoming apparent that more complex models of cancer are required to fully appreciate the contributing factors that drive tumorigenesis in vivo and increase the efficacy of novel therapies that make the transition from pre-clinical models to clinical trials.
Here we present a methodology for generating uniform and reproducible tumor spheroids that can be subjected to siRNA functional screening. These spheroids display many characteristics that are found in solid tumors that are not present in traditional two-dimension culture. We show that several commonly used breast cancer cell lines are amenable to this protocol. Furthermore, we provide proof-of-principle data utilizing the breast cancer cell line BT474, confirming their dependency on amplification of the epidermal growth factor receptor HER2 and mutation of phosphatidylinositol-4,5-biphosphate 3-kinase (PIK3CA) when grown as tumor spheroids. Finally, we are able to further investigate and confirm the spatial impact of these dependencies using immunohistochemistry.

Identifying novel drug targets that transition from pre-clinical testing to human trials is a scientific priority. To that end, here we describe a functional genomics approach for examining the impact of gene depletion on cancer cell line spheroids, which more appropriately model human cancers in vivo.

利用功能基因组筛选识别在肿瘤细胞球体培养潜在的新药物靶标

The identification of functional driver events in cancer is central to furthering our understanding of cancer biology and indispensable for the discovery ...

科研

癌症研究

Invasive Behavior of Human Breast Cancer Cells in Embryonic Zebrafish

In many cases, cancer patients do not die of a primary tumor, but rather because of metastasis. Although numerous rodent models are available for studying cancer metastasis in vivo, other efficient, reliable, low-cost models are needed to quickly access the potential effects of (epi)genetic changes or pharmacological compounds. As such, we illustrate and explain the feasibility of xenograft models using human breast cancer cells injected into zebrafish embryos to support this goal. Under the microscope, fluorescent proteins or chemically labeled human breast cancer cells are transplanted into transgenic zebrafish embryos, Tg (fli:EGFP), at the perivitelline space or duct of Cuvier (Doc) 48 h after fertilization. Shortly afterwards, the temporal-spatial process of cancer cell invasion, dissemination, and metastasis in the living fish body is visualized under a fluorescent microscope. The models using different injection sites, i.e., perivitelline space or Doc are complementary to one another, reflecting the early stage (intravasation step) and late stage (extravasation step) of the multistep metastatic cascade of events. Moreover, peritumoral and intratumoral angiogenesis can be observed with the injection into the perivitelline space. The entire experimental period is no more than 8 days. These two models combine cell labeling, micro-transplantation, and fluorescence imaging techniques, enabling the rapid evaluation of cancer metastasis in response to genetic and pharmacological manipulations.

Here, we describe xenograft zebrafish models using two different injection sites, i.e., perivitelline space and duct of Cuvier, to investigate the invasive behavior and to assess the intravasation and extravasation potential of human breast cancer cells, respectively.

在斑马鱼胚胎人乳腺癌细胞的侵袭行为

In many cases, cancer patients do not die of a primary tumor, but rather because of metastasis. Although numerous rodent models are available for studying ...

Live-imaging of Breast Epithelial Cell Migration After the Transient Depletion of TIP60

The wound-healing assay is efficient and one of the most economical ways to study cell migration in vitro. Conventionally, images are taken at the beginning and end of an experiment using a phase-contrast microscope, and the migration abilities of cells are evaluated by the closure of wounds. However, cell movement is a dynamic phenomenon, and a conventional method does not allow for tracking single-cell movement. To improve current wound-healing assays, we use live-cell imaging techniques to monitor cell migration in real time. This method allows us to determine the cell migration rate based on a cell tracking system and provides a clearer distinction between cell migration and cell proliferation. Here, we demonstrate the use of live-cell imaging in wound-healing assays to study the different migration abilities of breast epithelial cells influenced by the presence of TIP60. As cell motility is highly dynamic, our method provides more insights into the processes of wound healing than a snapshot of wound closure taken with the traditional imaging techniques used for wound-healing assays.

Here, we present the real-time monitoring of cell migration in a wound-healing assay using TIP60-depleted MCF10A breast epithelial cells. The implementation of live-cell imaging techniques in our protocol allows us to analyze and visualize single-cell movement in real time and across time.

TIP60 短暂耗竭后乳腺上皮细胞迁移的活体影像学研究

The wound-healing assay is efficient and one of the most economical ways to study cell migration in vitro. Conventionally, images are taken at the ...

Evaluating the Role of Mitochondrial Function in Cancer-related Fatigue

Fatigue is a common and debilitating condition that affects most cancer patients. To date, fatigue remains poorly characterized with no diagnostic test to objectively measure the severity of this condition. Here we describe an optimized method for assessing mitochondrial function of PBMCs collected from fatigued cancer patients. Using a compact extracellular flux system and sequential injection of respiratory inhibitors, we examined PBMC mitochondrial functional status by measuring basal mitochondrial respiration, spare respiratory capacity, and energy phenotype, which describes the preferred energy pathway to respond to stress. Fresh PBMCs are readily available in the clinical setting using standard phlebotomy. The entire assay described in this protocol can be completed in less than 4 hours without the involvement of complex biochemical techniques. Additionally, we describe a normalization method that is necessary for obtaining reproducible data. The simple procedure and normalization methods presented allow for repeated sample collection from the same patient and generation of reproducible data that can be compared between time points to evaluate potential treatment effects.

Our goal was to develop a practical protocol to evaluate mitochondrial dysfunction associated with fatigue in cancer patients. This innovative protocol is optimized for clinical use involving only standard phlebotomy and basic laboratory procedures.

线粒体功能在癌症相关疲劳中的作用评价

Fatigue is a common and debilitating condition that affects most cancer patients. To date, fatigue remains poorly characterized with no diagnostic test to ...

Establishment and Characterization of Three Afatinib-resistant Lung Adenocarcinoma PC-9 Cell Lines Developed with Increasing Doses of Afatinib

Acquired resistance to molecular target inhibitors is a severe problem in cancer therapy. Lung cancer remains the leading cause of cancer-related death in most countries. The discovery of "oncogenic driver mutations," such as epidermal growth factor receptor (EGFR)-activating mutations, and subsequent development of molecular targeted agents of EGFR tyrosine kinase inhibitors (TKIs) (gefitinib, erlotinib, afatinib, dacomitinib, and osimertinib) have dramatically altered lung cancer treatment in recent decades. However, these drugs are still not effective in patients with non-small cell lung cancer (NSCLC) carrying EGFR-activating mutations. Following acquired resistance, the systemic progression of NSCLC remains a significant obstacle in treating patients with EGFR mutation-positive NSCLC. Here, we present a stepwise dose escalation method for establishing three independent acquired afatinib-resistant cell lines from NSCLC PC-9 cells harboring EGFR-activating mutations of 15-base pair deletions in EGFR exon 19. Methods for characterizing the three independent afatinib-resistance cell lines are briefly presented. The acquired resistance mechanisms to EGFR TKIs are heterogeneous. Therefore, multiple cell lines with acquired resistance to EGFR-TKIs must be examined. Ten to twelve months are required to obtain cell lines with acquired resistance using this stepwise dose escalation approach. The discovery of novel acquired resistance mechanisms will contribute to the development of more effective and safe therapeutic strategies.

A method for establishing afatinib-resistance cell lines from lung adenocarcinoma PC-9 cells was developed, and resistant cells were characterized. The resistant cells can be used to investigate epidermal growth factor receptor tyrosine kinase inhibitor-resistance mechanisms, applicable for patients with non-small cell lung cancer.

建立和表征三个抗阿法替尼的肺腺癌PC-9细胞系与增加剂量的阿法替尼开发

Acquired resistance to molecular target inhibitors is a severe problem in cancer therapy. Lung cancer remains the leading cause of cancer-related death in ...

Differentiation of Mouse Breast Epithelial HC11 and EpH4 Cells

Cadherins play an important role in the regulation of cell differentiation as well as neoplasia. Here we describe the origins and methods of the induction of differentiation of two mouse breast epithelial cell lines, HC11 and EpH4, and their use to study complementary stages of mammary gland development and neoplastic transformation.
The HC11 mouse breast epithelial cell line originated from the mammary gland of a pregnant Balb/c mouse. It differentiates when grown to confluence attached to a plastic Petri dish surface in medium containing fetal calf serum and Hydrocortisone, Insulin and Prolactin (HIP medium). Under these conditions, HC11 cells produce the milk proteins &#946;-casein and whey acidic protein (WAP), similar to lactating mammary epithelial cells, and form rudimentary mammary gland-like structures termed &#34;domes&#34;.
The EpH4 cell line was derived from spontaneously immortalized mouse mammary gland epithelial cells isolated from a pregnant Balb/c mouse. Unlike HC11, EpH4 cells can fully differentiate into spheroids (also called mammospheres) when cultured under three-dimensional (3D) growth conditions in HIP medium. Cells are trypsinized, suspended in a 20% matrix consisting of a mixture of extracellular matrix proteins produced by Engelbreth-Holm-Swarm (EHS) mouse sarcoma cells, plated on top of a layer of concentrated matrix coating a plastic Petri dish or multiwell plate, and covered with a layer of 10% matrix-containing HIP medium. Under these conditions, EpH4 cells form hollow spheroids that exhibit apical-basal polarity, a hollow lumen, and produce &#946;-casein and WAP.
Using these techniques, our results demonstrated that the intensity of the cadherin/Rac signal is critical for the differentiation of HC11 cells. While Rac1 is necessary for differentiation and low levels of activated RacV12 increase differentiation, high RacV12 levels block differentiation while inducing neoplasia. In contrast, EpH4 cells represent an earlier stage in mammary epithelial differentiation, which is inhibited by even low levels of RacV12.

We describe techniques for differentiation induction of two breast epithelial lines, HC11 and EpH4. While both require fetal calf serum, insulin, and prolactin to produce milk proteins, EpH4 cells can fully differentiate into mammospheres in three-dimensional culture. These complementary models are useful for signal transduction studies of differentiation and neoplasia.

小鼠乳腺皮下HC11和EpH4细胞的分化

Cadherins play an important role in the regulation of cell differentiation as well as neoplasia. Here we describe the origins and methods of the induction ...

Studying Triple Negative Breast Cancer Using Orthotopic Breast Cancer Model

Triple-negative breast cancer (TNBC) is an aggressive breast cancer subtype with limited therapeutic options. When compared to patients with less aggressive breast tumors, the 5-year survival rate of TNBC patients is 77% due to their characteristic drug-resistant phenotype and metastatic burden. Toward this end, murine models have been established aimed at identifying novel therapeutic strategies limiting TNBC tumor growth and metastatic spread. This work describes a practical guide for the TNBC orthotopic model where MDA-MB-231 breast cancer cells suspended in a basement membrane matrix are implanted in the fourth mammary fat pad, which closely mimics the cancer cell behavior in humans. Measurement of tumors by caliper, lung metastasis assessment via in vivo and ex vivo imaging, and molecular detection are discussed. This model provides an excellent platform to study therapeutic efficacy and is especially suitable for the study of the interaction between the primary tumor and distal metastatic sites.

This work presets an advanced protocol to accurately assess tumor loading by detection of green fluorescent protein and bioluminescence signals as well as the integration of quantitative molecular detection technique.

研究使用正交乳腺癌模型的三阴性乳腺癌

Triple-negative breast cancer (TNBC) is an aggressive breast cancer subtype with limited therapeutic options. When compared to patients with less ...

Measuring the Motor Aspect of Cancer-Related Fatigue using a Handheld Dynamometer

Cancer-related fatigue (CRF) is commonly reported by patients both during and after receiving treatment for cancer. Current CRF diagnoses rely on self-report questionnaires which are subject to report and recall biases. Objective measurements using a handheld dynamometer, or handgrip device, have been shown in recent studies to correlate significantly with subjective self-reported fatigue scores. However, variations of both the handgrip fatigue test and fatigue index calculations exist in the literature. The lack of standardized methods limits the utilization of the handgrip fatigue test in the clinical and research settings. In this study, we provide detailed methods for administering the physical fatigue test and calculating the fatigue index. These methods should supplement existing self-reported fatigue questionnaires and help clinicians assess fatigue symptom severity in an objective and quantitative manner.

Simple and accessible methods were developed to measure the motor aspect of cancer-related fatigue objectively and quantitatively. We describe, in detail, ways to administer the physical fatigue test using a simple handgrip device as well as methods to calculate fatigue indices.

使用手持测功机测量癌症相关疲劳的马达方面

Cancer-related fatigue (CRF) is commonly reported by patients both during and after receiving treatment for cancer. Current CRF diagnoses rely on ...

Orthotopic Transplantation of Breast Tumors as Preclinical Models for Breast Cancer

Preclinical models that faithfully recapitulate tumor heterogeneity and therapeutic response are critical for translational breast cancer research. Immortalized cell lines are easy to grow and genetically modify to study molecular mechanisms, yet the selective pressure from cell culture often leads to genetic and epigenetic alterations over time. Patient-derived xenograft (PDX) models faithfully recapitulate the heterogeneity and drug response of human breast tumors. PDX models exhibit a relatively short latency after orthotopic transplantation that facilitates the investigation of breast tumor biology and drug response. The transplantable genetically engineered mouse models allow the study of breast tumor immunity. The current protocol describes the method to orthotopically transplant breast tumor fragments into the mammary fat pad followed by drug treatments. These preclinical models provide valuable approaches to investigate breast tumor biology, drug response, biomarker discovery and mechanisms of drug resistance.

Patient-derived xenograft (PDX) models and transplantable genetically engineered mouse models faithfully recapitulate human disease and are preferred models for basic and translational breast cancer research. Here, a method is described to orthotopically transplant breast tumor fragments into the mammary fat pad to study tumor biology and evaluate drug response.

乳腺癌临床前模型乳腺肿瘤的矫形移植

Preclinical models that faithfully recapitulate tumor heterogeneity and therapeutic response are critical for translational breast cancer research. ...

Modeling Breast Cancer in Human Breast Tissue using a Microphysiological System

Breast cancer (BC) remains a leading cause of death for women. Despite more than $700 million invested in BC research annually, 97% of candidate BC drugs fail clinical trials. Therefore, new models are needed to improve our understanding of the disease. The NIH Microphysiological Systems (MPS) program was developed to improve the clinical translation of basic science discoveries and promising new therapeutic strategies. Here we present a method for generating MPS for breast cancers (BC-MPS). This model adapts a previously described approach of culturing primary human white adipose tissue (WAT) by sandwiching WAT between adipose-derived stem cell sheets (ASC)s. Novel aspects of our BC-MPS include seeding BC cells into non-diseased human breast tissue (HBT) containing native extracellular matrix, mature adipocytes, resident fibroblasts, and immune cells; and sandwiching the BC-HBT admixture between HBT-derived ASC sheets. The resulting BC-MPS is stable in culture ex vivo for at least 14 days. This model system contains multiple elements of the microenvironment that influence BC including adipocytes, stromal cells, immune cells, and the extracellular matrix. Thus BC-MPS can be used to study the interactions between BC and its microenvironment. 
We demonstrate the advantages of our BC-MPS by studying two BC behaviors known to influence cancer progression and metastasis: 1) BC motility and 2) BC-HBT metabolic crosstalk. While BC motility has previously been demonstrated using intravital imaging, BC-MPS allows for high-resolution time-lapse imaging using fluorescence microscopy over several days. Furthermore, while metabolic crosstalk was previously demonstrated using BC cells and murine pre-adipocytes differentiated into immature adipocytes, our BC-MPS model is the first system to demonstrate this crosstalk between primary human mammary adipocytes and BC cells in vitro.

This protocol describes the construction of an in vitro microphysiological system for studying breast cancer using primary human breast tissue with off the shelf materials.

使用微生理系统模拟人体乳房组织中的乳腺癌

Breast cancer (BC) remains a leading cause of death for women. Despite more than $700 million invested in BC research annually, 97% of candidate BC drugs ...