This work was supported by the National Natural Science Foundation of China (Grant Number. 82174506).

This study was approved by the ethics committee of Yueyang Hospital, affiliated with Shanghai University of Traditional Chinese Medicine (reference no. 2021-062), and each participant signed an informed consent form.
1. Experiment preparations
<ol>
	<li>Camera settings:
	<ol>
		<li>Place three tripods in front of the operation table, and connect them with three cameras.</li>
		<li>Set the shooting parameters of the cameras as follows: resolution 1280 x720 pixels, format MP4, full manual mode (M), aperture F1.2, shutter 1/1000s, ISO 6400, automatic white balance, optical zoom 0mm. 
		NOTE: The angle between every two cameras is required to be set at 60&#176;-120&#176; (Figure 1A).</li>
	</ol>
	</li>
	<li>Tracking marker placement:
	<ol>
		<li>Attach seven reflective balls with a diameter of 6.5 mm on the holding-needle hand of each participant for video recording as detailed in steps 1.2.2-1.2.4 and shown in Figure 2A.</li>
		<li>Wrist: Attach one ball on the midpoint of the ulna and radial styloid defined as tracking point &#34;wrist joint &#34;(WJ)</li>
		<li>Thumb: Attach one ball each on the center of thumb nail defined as tracking point &#34;thumb tip&#34; (TT), the IP joint defined as tracking point &#34;thumb end joint&#34; (TEJ), and the metacarpophalangeal (MCP) joint defined as tracking point &#34;thumb base joint&#34; (TBJ), respectively.</li>
		<li>Forefinger: Attach one ball each on the center of the forefinger nail defined as tracking point &#34;forefinger tip&#34; (FT), the proximal interphalangeal (PIP) joint defined as tracking point &#34;forefinger middle joint&#34; (FMJ), and the MCP joint defined as tracking point &#34;forefinger base joint&#34; (FBJ), respectively.</li>
	</ol>
	</li>
</ol>
2. Video shooting and editing
<ol>
	<li>Place a small 15 cm x 15 cm x 15 cm 3D calibration frame with 8 points on the operating table for 3D calibration (Figure 1B,C).</li>
	<li>Remove the frame from the table after taking a video of the calibration frame for at least 8 s.</li>
	<li>Instruct the participants to perform AM on the acupuncture point LI11 (Quchi) of the volunteer, including lifting-thrusting and twirling skills, to control the needle to move up and down and rotate with thumb and forefinger, respectively. Take the videos of the above skills for at least 10 cycles. 
	NOTE: The inclusion and exclusion criteria of the participants to perform AM and volunteers to provide acupuncture points for needling are listed. Participant inclusion: (1) acupuncture teacher or student finished the &#34;lifting-thrusting skill&#34; and &#34;twirling skill&#34; chapter in the course textbook entitled &#39;Acupuncture and Moxibustion Techniques and Manipulations19; (2) participant should have hands-on needling experience with the human body for more than 5 times. Participant exclusion: (1) non-acupuncture teachers or students; (2) acupuncture students without any hands-on needling experience with the human body. Volunteer inclusion: (1) age between 16-60 years old; (2) no obvious skin damage, rupture, suppuration or obvious exudation around LI11 on the right arm. Volunteer exclusion: (1) individuals with a history of smoking, alcohol or drug abuse; (2) individuals with blood system diseases or obvious bleeding tendency; (3) individuals with chronic mental illness or mental disorders; (4) pregnant women; (5) individuals with a history of fainting needles.</li>
	<li>Export all the videos from the cameras to the designated disk of the computer. Rename the 3D calibration videos in cameras 1, 2, 3 as &#34;ca-1.mp4&#34;, &#34;ca-2.mp4&#34; and &#34;ca-3.mp4&#34;.</li>
	<li>Synchronize all manipulation videos in the video editing software (e.g., Adobe premiere pro) and export them named as &#34;lifting-thrusting-1.avi&#34;, &#34;lifting-thrusting-2.avi&#34;, &#34;lifting-thrusting-3.avi&#34;, &#34;twirling-1.avi&#34;, &#34;twirling-2.avi&#34; and &#34;twirling-3.avi&#34;, respectively. 
	&#8203;NOTE: Refer to Supplementary File 1 for the video synchronization instructions of the video editing software used in this study.</li>
</ol>
3. Project configuration of Simi Reality Motion System (motion capture and analysis software)
<ol>
	<li>Open the motion capture and analysis software and choose Create a New Project. Set the project name in Project Label and click Create and Save to save the project in the designated disk.</li>
	<li>Choose Specification &#62; Points &#62; Right/Left Hand and drag the above tracking points from the Predefined Points box into the Used Points box, then click on the Close button to continue. 
	NOTE: All the following steps take the tracking points of the right hand as an example.</li>
	<li>Choose Specification &#62; Connections and click on New Connection
	<ol>
		<li>Input connection name &#34;forefinger III right&#34;. Select &#34;forefinger middle joint right&#34; as the Starting Point and Line to the point &#34;forefinger tip right&#34; in the same window</li>
		<li>Click on the Apply and Close buttons to finish the establishment of the connection.</li>
	</ol>
	</li>
	<li>Add and rename the camera groups
	<ol>
		<li>Right-click on Cameras &#62; Add Camera Group to add new camera groups.</li>
		<li>Right-click on Cameras &#62; Rename to rename the camera groups as &#34;lifting-thrusting camera group&#34; and &#34;twirling camera group&#34;, respectively.</li>
	</ol>
	</li>
	<li>Right-click on the Lifting-Thrusting Camera Group &#62; Add Camera
	<ol>
		<li>Click on the Select File button in the Tracking box.</li>
		<li>Click on Open Existing File and select the operation video &#34;lifting-thrusting-1.avi&#34; in the next window, then click on Apply to finish video import.</li>
		<li>Similar to the above actions, click on Select File in the 3D Calibration box, and import the corresponding calibration video &#34;ca-1.mp4&#34;.</li>
	</ol>
	</li>
	<li>According to step 3.5, continue to import the operation videos &#34;lifting-thrusting-2.avi&#34; and &#34;lifting-thrusting-3.avi&#34;, and their corresponding calibration videos &#34;ca-2.mp4&#34; and &#34;ca-3.mp4&#34; in the Lifting-Thrusting Camera Group, respectively. 
	NOTE: There should be 3 cameras in the Lifting-Thrusting Camera Group in the project window after sections 3.4 and 3.5.</li>
	<li>According to steps 3.4, 3.5, and 3.6, import the twirling skill and calibration videos into the Twirling Camera Group.</li>
</ol>
4. Video analysis
<ol>
	<li>3D calibration for each camera
	<ol>
		<li>Expand Lifting-Thrusting Camera Group and right-click on Lifting-Thrusting-1 &#62; Properties.</li>
		<li>Click on the 3D Calibration button in the 3D Calibration box; input the description and add 8 points by clicking on Add Point button for 8 times</li>
		<li>Click on Apply after setting the name and corresponding X, Y, Z value for each point according to the calibration parameters (Table 1).</li>
		<li>After configuring all points, move the mouse to click each endpoint of the calibration video to finish the 3D calibration.</li>
		<li>Follow steps 4.1.1-4.1.4 to complete the 3D calibration of the other cameras in the same group and the cameras in the Twirling Camera Group.</li>
	</ol>
	</li>
	<li>3D finger motion tracking
	<ol>
		<li>Right-click on Lifting-Thrusting Camera Group &#62; 3D Tracking, select all the cameras, and click on the OK button to open the 3D tracking window.</li>
		<li>Set the Track Using Pattern Matching (all points) for all the cameras and manually click on all the tracking points in the first frame.</li>
		<li>Click on the Search Automatically button to start automatic 3D tracking frame by frame.</li>
		<li>Follow steps 4.2.1-4.2.3 to complete the motion tracking of the Twirling Camera Group. 
		NOTE: If a tracking point is lost during the automatic 3D tracking, select the lost point line, right-click on Discard Point From Here, then re-click the point and the Search Automatically button. Select Yes if the message &#34;No start frame for tracking has been set for 3 selected camera(s). it can be set individually in camera properties. Do you want to set start frame to frame 0 for all cameras without start frame and continue now?&#34; pops up.</li>
	</ol>
	</li>
	<li>Data export
	<ol>
		<li>Right-click on Lifting-Thrusting Camera Group &#62; New 3D Calculation, select all the cameras, and check Update Data Continuously and Store Data Explicitly in File in Create 3D Data window. Click the OK button to continue.</li>
		<li>Right-click on the folder Lifting-Thrusting-3D Coordinates Data &#62; Export, check Column Headings, Tracking Names, Start Time and Frequency, Time Information in First Column, X, Y, Z, v(X), v(Y), v(Z) in the Export window</li>
		<li>Click the Export button to export the data file (*.txt) with the customized name. Export the data file of the Twirling Camera Group in the same way.</li>
	</ol>
	</li>
</ol>
5. Data analysis
NOTE: An original PHP script is used to browse and analyze the data files exported by the motion capture and analysis software. All the source code has been shared in a GitHub repository20.
<ol>
	<li>After the data files exported from the motion capture and analysis software are uploaded to a specific server folder running this script, open the script and input the Username and Password to log in.</li>
	<li>Click on Add New Participant, select the Participant Type and Gender, and input the Participant Name, Age, and Practice Time in the pop-up page; click on Submit to finish adding a new participant.</li>
	<li>Click on Add New Record corresponding to the newly added participant in the list page, then input the Folder Name containing the uploaded data files of the motion capture and analysis software and select the Operation Date; click on Submit to continue.</li>
	<li>Click on Analysis corresponding to the newly added operation record, then select Skill and click on Submit. The script will identify and display all the valid crests and troughs for manual review. 
	NOTE: A certain crest or trough can be reselected manually in the corresponding drop-down list if the script incorrectly identifies it. Based on these crests and troughs, the average values of amplitudes and velocities along three axes of each tracking point and the operating time of lifting, thrusting, twirling left, and twirling right actions can be calculated and displayed by the script. The calculation method of these parameters is shown in Figure 3.</li>
</ol>

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The authors have nothing to disclose.

This study established the measurement method of the kinematic parameters of AM in vivo and obtained the data of motion amplitude, velocity, and operation time of the six important tracking points on the thumb and forefinger along three axes. Meanwhile, based on the 3D calibration frame, a 3D stick view and corresponding animation of the thumb and forefinger during needling were generated. The thumb and forefinger movement of AM can be fully displayed with the synchronous playback of kinematic parameter curve an...

As a kind of the clinical skills of traditional Chinese medicine (TCM) and physical stimulation, acupuncture manipulation (AM) is often regarded as an important factor that affects the therapeutic effect of acupuncture1,2. Many studies have confirmed that different AM or different stimulation parameters (needling velocity, amplitude, frequency, etc.) of the same AM resulted in different therapeutic effects3,4,5,6,7. Therefore, the measurement of relevant kinematic parameters of AM and correlation analysis with the therapeutic effect can provide useful data support and reference for the clinical treatment with acupuncture8,9.
The measurement of kinematic parameters of the AM began in the 1980s10. In the early days, the electrical signal conversion technology based on variable resistance was mainly used to convert the displacement signal of the needle body into a voltage or current signal for displaying and recording the amplitude and frequency data of AM11. Moreover, the famous ATP-II Chinese medicine acupuncture technique tester II (ATP-II) with this technology currently has been used by many traditional Chinese medicine universities of China12. After that, with the continuous development and innovation of sensor technology, different types of sensors were used to collect kinematic parameters of AM. For example, the three axes electromagnetic motion sensor was attached to the needle handle to acquire needling amplitude and velocity13; the bioelectric signal sensor was placed on the dorsal horn of the animal&#39;s spinal cord to record needling frequency14, etc. Although the quantitative research of AM based on the above two types of technologies has completed the acquisition of relevant kinematic parameters during needling, its main disadvantages are the inability to perform the real-time non-invasive measurement and the change of operating feel caused by the modification of the needle body.
In recent years, motion tracking technology was gradually applied to the quantitative research of AM15,16. Because it is based on the frame-by-frame analysis of needling video, the measurement of acupuncture parameters can be acquired during in vivo operation without modifying the needle body. This technology has been used to measure the kinematic parameters such as amplitude, velocity, acceleration, and frequency of four tracking points of thumb and forefinger during needling in a two-dimensional (2D) plane and established the corresponding finger stick figure15. Some studies also measured the angle change range of interphalangeal (IP) joint of thumb and forefinger with similar technology9,17,18. However, the current studies on AM analysis are still mainly limited to the 2D motion plane, and the number of tracking points is relatively small. So far, there is no complete three-dimensional (3D) kinematics measurement and analysis method for AM, and no related data was published.
To solve the above problems, this study will use 3D motion tracking technology to measure the kinematic parameters of the seven tracking points of hand during needling. This protocol aims to provide a complete technical solution for the kinematic analysis on AM, as well as the further study on the dose-effect correlation of acupuncture.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>3D calibration frame</td>
			<td>Any brand</td>
			<td></td>
			<td>15 x 15 x 15 cm</td>
		</tr>
		<tr>
			<td>Acupuncture needles</td>
			<td>Suzhou Medical Appliance Factory</td>
			<td></td>
			<td>0.35 x 40 mm</td>
		</tr>
		<tr>
			<td>Double-sided tape</td>
			<td>Any brand</td>
			<td></td>
			<td>Round, 1 cm-diameter</td>
		</tr>
		<tr>
			<td>Reflective balls</td>
			<td>Simi Reality Motion Systems GmbH</td>
			<td></td>
			<td>6.5 mm-diameter</td>
		</tr>
		<tr>
			<td>SD card</td>
			<td>Western Digital Corporation</td>
			<td></td>
			<td>SDXC UHS-I</td>
		</tr>
		<tr>
			<td>SD card reader</td>
			<td>UGREEN Group Limited</td>
			<td></td>
			<td>USB 3.0</td>
		</tr>
		<tr>
			<td>Simi Motion</td>
			<td>Simi Reality Motion Systems GmbH</td>
			<td></td>
			<td>Ver.8.5.15</td>
		</tr>
		<tr>
			<td>Swab</td>
			<td>Any brand</td>
			<td></td>
			<td>The volume fraction of ethanol is 70%-80%</td>
		</tr>
		<tr>
			<td>Three cameras</td>
			<td>Victor Company of Japan, Limited</td>
			<td></td>
			<td>JVC GC-PX100BAC</td>
		</tr>
		<tr>
			<td>Three tripods</td>
			<td>Any brand</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

three-dimensional finger motion tracking during needling: a solution for the kinematic analysis of acupuncture manipulation

Three-dimensional (3D) motion tracking has been used in many fields, such as the researches of sport and medical skills. This experiment aimed to use 3D motion tracking technology to measure the kinematic parameters of the joints of fingers during acupuncture manipulation (AM) and establish three technical indicators "amplitude, velocity and time". This method can reflect the operation characteristics of AM and provide quantitative parameters along three axes of multiple finger joints. The current evidence shows that the method has great potential for future applications such as the study of the dose-effect relationship of acupuncture, teaching, and learning of AM, and the measurement and preservation of famous acupuncturists' AM.

After establishing this experimental method, the lifting-thrusting and twirling skills of basic AM of nineteen acupuncture teachers from the School of Acupuncture-Moxibustion and Tuina of Shanghai University of TCM were measured using 3D motion tracking. According to the definition of a joint coordinate system (JCS) for the shoulder, elbow, wrist, and hand proposed by the Standardization and Terminology Committee (STC) of the International Society of Biomechanics21, seven finger tracking points ha...

Watch this Scientific Journal Video about Three-Dimensional Finger Motion Tracking during Needling: A Solution for the Kinematic Analysis of Acupuncture Manipulation at JoVE.com

Three-Dimensional Finger Motion Tracking during Needling: A Solution for the Kinematic Analysis of Acupuncture Manipulation

This experimental method describes a solution for the kinematic analysis of acupuncture manipulation with three-dimensional finger motion tracking technology.

Three-dimensional (3D) motion tracking has been used in many fields, such as the researches of sport and medical skills. This experiment aimed to use 3D ...

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2.7K Views.  Shanghai University of Traditional Chinese Medicine. This experimental method describes a solution for the kinematic analysis of acupuncture manipulation with three-dimensional finger motion tracking technology.

Video: Three-Dimensional Finger Motion Tracking during Needling: A Solution for the Kinematic Analysis of Acupuncture Manipulation

Heterotypic Three-dimensional In Vitro Modeling of Stromal-Epithelial Interactions During Ovarian Cancer Initiation and Progression

Epithelial ovarian cancers (EOCs) are the leading cause of death from gynecological malignancy in Western societies. Despite advances in surgical treatments and improved platinum-based chemotherapies, there has been little improvement in EOC survival rates for more than four decades 1,2. Whilst stage I tumors have 5-year survival rates &gt;85%, survival rates for stage III/IV disease are &lt;40%. Thus, the high rates of mortality for EOC could be significantly decreased if tumors were detected at earlier, more treatable, stages 3-5. At present, the molecular genetic and biological basis of early stage disease development is poorly understood. More specifically, little is known about the role of the microenvironment during tumor initiation; but known risk factors for EOCs (e.g. age and parity) suggest that the microenvironment plays a key role in the early genesis of EOCs. We therefore developed three-dimensional heterotypic models of both the normal ovary and of early stage ovarian cancers. For the normal ovary, we co-cultured normal ovarian surface epithelial (IOSE) and normal stromal fibroblast (INOF)&nbsp;cells, immortalized by retrovrial transduction of the catalytic subunit of human telomerase holoenzyme (hTERT) to extend the lifespan of these cells in culture. To model the earliest stages of ovarian epithelial cell transformation, overexpression of the CMYC oncogene in IOSE cells, again co-cultured with INOF cells. These heterotypic models were used to investigate the effects of aging and senescence on the transformation and invasion of epithelial cells. Here we describe the methodological steps in development of these three-dimensional model; these methodologies aren't specific to the development of normal ovary and ovarian cancer tissues, and could be used to study other tissue types where stromal and epithelial cell interactions are a fundamental aspect of the tissue maintenance and disease development.

Heterotypic Three-dimensional In Vitro Modeling of Stromal-Epithelial Interactions During Ovarian Cancer Initiation and Progression

We describe methodologies for establishing in vitro heterotypic three-dimensional models comprising ovarian fibroblasts and normal ovarian surface or ovarian cancer epithelial cells. We discuss the use of these models to study stromal-epithelial interactions that occur during ovarian cancer development.

Epithelial ovarian cancers (EOCs) are the leading cause of death from gynecological malignancy in Western societies. Despite advances in surgical ...

Human Neuroendocrine Tumor Cell Lines as a Three-Dimensional Model for the Study of Human Neuroendocrine Tumor Therapy

Neuroendocrine tumors (NETs) are rare tumors, with an incidence of two per 100,&#x200A;000 individuals per year, and they account for 0.5% of all human malignancies.1 Other than surgery for the minority of patients who present with localized disease, there is little or no survival benefit of systemic therapy. Therefore, there is a great need to better understand the biology of NETs, and in particular define new therapeutic targets for patients with nonresectable or metastatic neuroendocrine tumors. 3D cell culture is becoming a popular method for drug screening due to its relevance in modeling the in vivo tumor tissue organization and microenvironment.2,3 The 3D multicellular spheroids could provide valuable information in a more timely and less expensive manner than directly proceeding from 2D cell culture experiments to animal (murine) models.
To facilitate the discovery of new therapeutics for NET patients, we have developed an in vitro 3D multicellular spheroids model using the human NET cell lines. The NET cells are plated in a non-adhesive agarose-coated 24-well plate and incubated under physiological conditions (5% CO2, 37 &deg;C) with a very slow agitation for 16-24 hr after plating. The cells form multicellular spheroids starting on the 3rd or 4th day. The spheroids become more spherical by the 6th day, at which point the drug treatments are initiated. The efficacy of the drug treatments on the NET spheroids is monitored based on the morphology, shape and size of the spheroids with a phase-contrast light microscope. The size of the spheroids is estimated automatically using a custom-developed MATLAB program based on an active contour algorithm. Further, we demonstrate a simple method to process the HistoGel embedding on these 3D spheroids, allowing the use of standard histological and immunohistochemical techniques. 
This is the first report on generating 3D spheroids using NET cell lines to examine the effect of therapeutic drugs. We have also performed histology on these 3D spheroids, and displayed an example of a single drug's effect on growth and proliferation of the NET spheroids. Our results support that the NET spheroids are valuable for further studies of NET biology and drug development.

We present a simple agarose overlay platform to grow 3D multicellular spheroids using neuroendocrine cancer cell lines. This method provides a very convenient way to examine the effect of therapeutic drugs on the neuroendocrine tumor cells. It could also help us establish human neuroendocrine tumor spheroids for cancer therapy.

Neuroendocrine tumors (NETs) are rare tumors, with an incidence of two per 100,&#x200A;000 individuals per year, and they account for 0.5% of all human ...

Three-dimensional Co-culture Model for Tumor-stromal Interaction

Cancer progression (initiation, growth, invasion and metastasis) occurs through interactions between malignant cells and the surrounding tumor stromal cells. The tumor microenvironment is comprised of a variety of cell types, such as fibroblasts, immune cells, vascular endothelial cells, pericytes and bone-marrow-derived cells, embedded in the extracellular matrix (ECM). Cancer-associated fibroblasts (CAFs) have a pro-tumorigenic role through the secretion of soluble factors, angiogenesis and ECM remodeling. The experimental models for cancer cell survival, proliferation, migration, and invasion have mostly relied on two-dimensional monocellular and monolayer tissue cultures or Boyden chamber assays. However, these experiments do not precisely reflect the physiological or pathological conditions in a diseased organ. To gain a better understanding of tumor stromal or tumor matrix interactions, multicellular and three-dimensional cultures provide more powerful tools for investigating intercellular communication and ECM-dependent modulation of cancer cell behavior. As a platform for this type of study, we present an experimental model in which cancer cells are cultured on collagen gels embedded with primary cultures of CAFs.

Here we present a protocol to co-culture in three-dimensions, which is useful for investigating multicellular interactions and extracellular matrix-dependent modulation of cancer cell behavior. In this experimental model, cancer cells are cultured on collagen gels embedded with human cancer-associated fibroblasts.

Cancer progression (initiation, growth, invasion and metastasis) occurs through interactions between malignant cells and the surrounding tumor stromal ...

Three-Dimensional (3D) Tumor Spheroid Invasion Assay

Invasion of surrounding normal tissues is generally considered to be a key hallmark of malignant (as opposed to benign) tumors. For some cancers in particular (e.g., brain tumors such as glioblastoma multiforme and squamous cell carcinoma of the head and neck &#8211; SCCHN) it is a cause of severe morbidity and can be life-threatening even in the absence of distant metastases. In addition, cancers which have relapsed following treatment unfortunately often present with a more aggressive phenotype. Therefore, there is an opportunity to target the process of invasion to provide novel therapies that could be complementary to standard anti-proliferative agents. Until now, this strategy has been hampered by the lack of robust, reproducible assays suitable for a detailed analysis of invasion and for drug screening. Here we provide a simple micro-plate method (based on uniform, self-assembling 3D tumor spheroids) which has great potential for such studies. We exemplify the assay platform using a human glioblastoma cell line and also an SCCHN model where the development of resistance against targeted epidermal growth factor receptor (EGFR) inhibitors is associated with enhanced matrix-invasive potential. We also provide two alternative methods of semi-automated quantification: one using an imaging cytometer and a second which simply requires standard microscopy and image capture with digital image analysis.

Three-Dimensional 3D Tumor Spheroid Invasion Assay

Invasion of surrounding normal tissues is a defining characteristic of malignant tumors. We provide here a simple, semi-automated micro-plate assay of invasion into a natural 3D biomatrix that has been exemplified with a number of models of advanced human cancers.

Invasion of surrounding normal tissues is generally considered to be a key hallmark of malignant (as opposed to benign) tumors. For some cancers in ...

Adapted Resistance Training Improves Strength in Eight Weeks in Individuals with Multiple Sclerosis

Hip weakness is a common symptom affecting walking ability in people with multiple sclerosis (MS). It is known that resistance strength training (RST) can improve strength in individuals with MS, however; it remains unclear the duration of RST that is needed to make strength gains and how to adapt hip strengthening exercises for individuals of varying strength using only resistance bands. This paper describes the methodology to set up and implement an adapted resistance strength training program, using resistance bands, for individuals with MS. Directions for pre- and post-strength tests to evaluate efficacy of the strength-training program are included. Safety features and detailed instructions outline the weekly program content and progression. Current evidence is presented showing that significant strength gains can be made within 8 weeks of starting a RST program. Evidence is also presented showing that resistance strength training can be successfully adapted for individuals with MS of varying strength with little equipment.

Hip weakness is a common symptom affecting walking ability in people with multiple sclerosis. Isolated muscle strengthening is a useful method to target specific weaknesses. This protocol describes a progressive resistance-training program using exercise bands to increase hip muscle strength.

Hip weakness is a common symptom affecting walking ability in people with multiple sclerosis (MS). It is known that resistance strength training (RST) can ...

Three-dimensional Alginate-bead Culture of Human Pituitary Adenoma Cells

A three-dimensional culture method is described in which primary pituitary adenoma cells are grown in alginate beads. Alginate is a polymer derived from brown sea algae. Briefly, the tumor tissue is cut into small pieces and submitted to an enzymatic digestion with collagenase and trypsin. Next, a cell suspension is obtained. The tumor cell suspension is mixed with 1.2% sodium alginate and dropped into a CaCl2 solution, and the alginate/cell suspension is gelled on contact with the CaCl2 to form spherical beads. The cells embedded in the alginate beads are supplied with nutrients provided by the culture media enriched with 20% FBS. Three-dimensional culture in alginate beads maintains the viability of adenoma cells for long periods of time, up to four months. Moreover, the cells can be liberated from the alginate by washing the beads with sodium citrate and seeded on glass coverslips for further immunocytochemical analyses. The use of a cell culture model allows for the fixation and visualization of the actin cytoskeleton with minimal disorganization. In summary, alginate beads provide a reliable culture system for the maintenance of pituitary adenoma cells.

Three-dimensional culture in alginate beads, due to its simplicity and reproducibility, was chosen to maintain the pituitary adenoma cells. The procedures included an initial enzymatic digestion and mechanical dissociation of the tumor tissue, and the subsequent cell suspension was encapsulated in alginate beads.

A three-dimensional culture method is described in which primary pituitary adenoma cells are grown in alginate beads. Alginate is a polymer derived from ...

Three-Dimensional Printing of a Complex Aortic Anomaly

Complex congenital aortic anomalies include diverse types of malformations that may be clinically asymptomatic or present with respiratory or esophageal symptoms. These anomalies may be associated with other congenital heart diseases. It is hard to identify the accurate anatomic vessel location from two-dimensional imaging data, such as computed tomography. As an additive manufacturing method, three-dimensional (3-D) printing can covert the acquired imaging data into 3-D physical models. This protocol describes the procedure for modeling the volumetric DICOM imaging into 3-D data and printing it as an anatomically realistic 3-D model. Using this model, surgeons can identify the vessel location of complex aortic anomalies, which is helpful for pre-operative planning and intra-operative guidance.

Here, we present a protocol to use three dimensional printed models for pre-operative planning and intra-operative reorganization of complicated vascular locations when handling a congenital aortic anomaly.

Complex congenital aortic anomalies include diverse types of malformations that may be clinically asymptomatic or present with respiratory or esophageal ...

Whole-Kidney Three-Dimensional Staining with CUBIC

Although conventional pathology provided numerous information about kidney microstructure, it was difficult to know the precise structure of blood vessels, proximal tubules, collecting ducts, glomeruli, and sympathetic nerves in the kidney due to the lack of three-dimensional (3D) information. Optical clearing is a good strategy to overcome this big hurdle. Multiple cells in a whole organ can be analyzed at single-cell resolution by combining tissue clearing and 3D imaging technique. However, cell labeling methods for whole-organ imaging remain underdeveloped. In particular, whole-mount organ staining is challenging because of the difficulty in antibody penetration. The present protocol developed a whole-mount mouse kidney staining for 3D imaging with the CUBIC (Clear, Unobstructed Brain/Body Imaging Cocktails and Computational analysis) tissue clearing method. The protocol has enabled visualizing renal sympathetic denervation after ischemia-reperfusion injury and glomerulomegaly in the early stage of diabetic kidney disease from a comprehensive viewpoint. Thus, this technique can lead to new discoveries in kidney research by providing a macroscopic perspective.

The present protocol describes a tissue clearing method and whole-mount immunofluorescent staining for three-dimensional (3D) kidney imaging. This technique can offer macroscopic perspectives in kidney pathology, leading to new biological discoveries.

Although conventional pathology provided numerous information about kidney microstructure, it was difficult to know the precise structure of blood ...

FSN Therapy for Knee Osteoarthritis Pain Management

Fu's Subcutaneous Needling for Knee Osteoarthritis Pain

Fu's subcutaneous needling (FSN) is a new acupuncture and dry needling technique based on traditional Chinese medicine. It rapidly produces long-lasting effects in soft tissue injuries, particularly in painful musculoskeletal conditions, by providing stimulation primarily in the subcutaneous area. Osteoarthritis (OA) is the most common joint disease in adults worldwide and is often accompanied by a painful syndrome of structural changes in the peripheral joints of the knee. However, the etiology of OA pain is not fully understood, though myofascial trigger points (MTrPs) are commonly found in the lower limb muscles (so-called "tightened muscles") of patients with knee OA. 
FSN has been used in many fields for the treatment of acute pain problems and can relieve muscle contraction from MTrPs, thereby improving the local circulation. This study recruited patients with pain from knee OA into an FSN group or a transcutaneous electrical nerve stimulation (TENS) group with three treatment sessions and a follow-up over the course of 2 weeks. The results showed that FSN was effective in treating soft tissue pain around the knee with OA. This study aimed to establish and visualize three key technical indicators during FSN therapy, including the FSN needle insertion point and layer; the frequency and duration of the swaying movement; and the manipulation of the reperfusion approach. These findings have great potential for future applications in myofascial pain treatment, especially for pain management. Following this protocol could enhance FSN skills.

Author Spotlight: Fu's Subcutaneous Needling for Knee Osteoarthritis Pain  

We present a protocol for using Fu's subcutaneous needling for knee osteoarthritis pain, which combines swaying movement and reperfusion approach techniques. This protocol has great potential for future applications in myofascial pain treatment and could enhance Fu's subcutaneous needling (FSN) manipulation skills.

Fu's subcutaneous needling (FSN) is a new acupuncture and dry needling technique based on traditional Chinese medicine. It rapidly produces long-lasting ...

Computer-Aided Three-Dimensional Visualization in the Treatment of Locally Advanced Thyroid Cancer

The diagnosis and treatment of locally advanced thyroid carcinoma are challenging. The challenge lies in the evaluation of the tumor scope and the formulation of an individualized treatment plan. Three-dimensional (3D) visualization has a wide range of applications in the field of medicine, although there are limited applications in thyroid cancer. We previously applied 3D visualization for the diagnosis and treatment of thyroid cancer. Through data collection, 3D modeling, and preoperative evaluation, we can obtain 3D information regarding the tumor outline, determine the extent of tumor invasion, and conduct adequate preoperative preparation and surgical risk assessment. This study aimed to demonstrate the feasibility of 3D visualization in locally advanced thyroid cancer. Computer-aided 3D visualization can be an effective method for accurate preoperative evaluation, the development of surgical methods, shortening the surgical time, and reducing the surgical risks. Furthermore, it can contribute to medical education and doctor-patient communication. We believe that the application of 3D visualization technology can improve outcomes and quality of life in patients with locally advanced thyroid cancer.

In diagnosing and treating locally advanced thyroid cancer, the application of computer-aided three-dimensional reconstruction can provide additional information regarding the tumor scope and anatomic characteristics, thereby assisting in risk assessment and surgical planning.

Three-Dimensional Finger Motion Tracking during Needling: A Solution for the Kinematic Analysis of Acupuncture Manipulation

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Heterotypic Three-dimensional In Vitro Modeling of Stromal-Epithelial Interactions During Ovarian Cancer Initiation and Progression

Human Neuroendocrine Tumor Cell Lines as a Three-Dimensional Model for the Study of Human Neuroendocrine Tumor Therapy

Three-dimensional Co-culture Model for Tumor-stromal Interaction

Three-Dimensional (3D) Tumor Spheroid Invasion Assay

Adapted Resistance Training Improves Strength in Eight Weeks in Individuals with Multiple Sclerosis

Three-dimensional Alginate-bead Culture of Human Pituitary Adenoma Cells

Three-Dimensional Printing of a Complex Aortic Anomaly

Whole-Kidney Three-Dimensional Staining with CUBIC

Fu's Subcutaneous Needling for Knee Osteoarthritis Pain

Computer-Aided Three-Dimensional Visualization in the Treatment of Locally Advanced Thyroid Cancer