This work was supported by the National Nature Foundation of China (grant no.82004504), the National Key Research and Development Program of the Ministry of Science and Technology of China (grant no.2018YFC1707606), Chinese Medicine Administration of Sichuan Province (grant no.2021MS199) and National Nature Foundation of China (grant no.82174236).

This study has been approved by the National Natural Science Foundation of China project, Constructing Dynamic Change rules of TCM Facial image Based on Association Analysis. The ethics approval number is 2021KL-027, and the ethics committee has approved the clinical study to be carried out in accordance with the approved documents which include clinical research protocol (2021.04.12, V2.0), informed consent (2021.04.12, V2.0), subject recruitment materials (2021.04.12, V2.0), study cases and/or case reports, subject dia...

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Based on the comparison results presented above, it is evident that the characteristics of the four algorithms under consideration are varied, and their distinct advantages and disadvantages are described below. The U-Net structure, based on the modification and expansion of a full convolution network, can obtain contextual information and precise positioning through a contracting path and a symmetrical expanding path. By classifying each pixel point, this algorithm achieves a higher segmentation accuracy and segments th...

Tongue observation is a widely utilized technique in traditional Chinese ethnic medicine (TCM). The color and shape of the tongue can reflect the physical condition and various disease properties, severities, and prognoses. For instance, in traditional Hmong medicine, the tongue&#39;s color is used to identify body temperature e.g., a red or purple tongue indicates pathological factors related to heat. In Tibetan medicine, a condition is judged by observing the tongue of a patient, paying attention to the color, shape, and moisture of the mucus. For instance, the tongues of patients with Heyi disease become red and rough or black and dry1; patients with Xieri disease2 have yellow and dry tongues; meanwhile, patients with Badakan disease3 have a white, humid, and soft tongue4. These observations reveal the close relationship between tongue features and physiology and pathology. Overall, the state of the tongue plays a vital role in diagnosis, disease identification, and evaluation of the treatment effect.
Simultaneously, owing to diverse living conditions and dietary practices among different ethnic groups, variations in tongue images are evident. The Lab model, established on the basis of an international standard for the determination of color, was formulated by the Commission International Eclairage (CIE) in 1931. In 1976, a color pattern was modified and named. The Lab color model is composed of three elements: L corresponds to brightness, while a and b are two color channels. a includes colors from dark green (low brightness value) to gray (medium brightness value) to bright pink (high brightness value); b goes from bright blue (low brightness value) to gray (medium brightness value) to yellow (high brightness value). By comparing the L x a x b values of the tongue color of five ethnic groups, Yang et al.5 found that the characteristics of tongue images of the Hmong, Hui, Zhuang, Han, and Mongolian groups were significantly distinct from each other. For example, the Mongolians have dark tongues with a yellow tongue coating, while the Hmong have light tongues with a white tongue coating, suggesting that tongue features can be used as a diagnostic indicator for assessing the health status of a population. Moreover, tongue images can function as an evaluation index for evidence-based medicine in clinical research of ethnic medicine. He et al.6 employed tongue images as a foundation for TCM diagnosis and systematically evaluated the safety and efficacy of Chou-Ling-Dan pellets (CLD granules-used to treat inflammatory and febrile diseases, including seasonal influenza in TCM) combined with Chinese and Western medicine. The results established the scientific validity of tongue images as an evaluation index for clinical studies. Nevertheless, traditional medical practitioners generally rely on subjectivity to observe tongue characteristics and assess patients&#39; physiological and pathological conditions, requiring more precise indicators.
The emergence of the internet and artificial intelligence technology has paved the way for digitizing and objectifying tongue diagnosis. This process involves using mathematical models to provide a qualitative and objective description of tongue images7, reflecting the content of the tongue image. The process includes several steps: image acquisition, optical compensation, color correction, and geometric transformation. The pre-processed images are then fed into an algorithmic model for image positioning and segmentation, feature extraction, pattern recognition, etc. The output of this process is a highly efficient and precise diagnosis of tongue image data, thereby achieving the goal of objectification, quantification, and informatization of tongue diagnosis8. Thus, the purpose of high efficiency and high precision processing of tongue diagnosis data is achieved. Based on tongue diagnosis knowledge and deep learning technology, this study automatically separated the tongue body and tongue coating from tongue images using a computer algorithm, in order to extract the quantitative features of tongues for doctors, improve the reliability and consistency of diagnosis, and provide methods for subsequent tongue diagnosis objectification research9.

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			<td>CPU</td>
			<td>Intel(R) Core(TM) i7-9700K</td>
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			<td>GPU</td>
			<td>&#160;NVIDIA GeForce RTX 3070 Ti (8192MB)</td>
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			<td>Microsoft Windows 10 Professional Edition (64-bit)</td>
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			<td>Python</td>
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objectification of tongue diagnosis in traditional medicine, data analysis, and study application

Tongue diagnosis is an essential technique of traditional Chinese medicine (TCM) diagnosis, and the need for objectifying tongue images through image processing technology is growing. The present study provides an overview of the progress made in tongue objectification over the past decade and compares segmentation models. Various deep learning models are constructed to verify and compare algorithms using real tongue image sets. The strengths and weaknesses of each model are analyzed. The findings indicate that the U-Net algorithm outperforms other models regarding precision accuracy (PA), recall, and mean intersection over union (MIoU) metrics. However, despite the significant progress in tongue image acquisition and processing, a uniform standard for objectifying tongue diagnosis has yet to be established. To facilitate the widespread application of tongue images captured using mobile devices in tongue diagnosis objectification, further research could address the challenges posed by tongue images captured in complex environments.

For the comparison results, see Figure 12, Figure 13, and Table 1, where the environment constructed by this study uses the same samples to train and test the algorithm model. MIoU indicator: U-Net &#62; Seg-Net &#62; PSPNet &#62; DeeplabV3; MPA indicator: U-Net &#62; Seg-Net &#62; PSPNet &#62; DeeplabV3; precision indicator: U-Net &#62; Seg-Net &#62; DeeplabV3 &#62; PSPNet; recall: U-Net &#62; Seg-Net &#62; PSPNet &#62; DeeplabV3. The larger th...

Watch this Scientific Journal Video about Objectification of Tongue Diagnosis in Traditional Medicine, Data Analysis, and Study Application at JoVE.com

Objectification of Tongue Diagnosis in Traditional Medicine, Data Analysis, and Study Application

The present study employed U-Net and other deep learning algorithms to segment a tongue image and compared the segmentation results to investigate the objectification of tongue diagnosis.

Tongue diagnosis is an essential technique of traditional Chinese medicine (TCM) diagnosis, and the need for objectifying tongue images through image ...

This protocol uses deep learning algorithm to segment tongue images and automatically separates the tongue from the whole image, which can be used for further research. This protocol compares the performance of four-tongue image segmentation algorithm to select the base algorithm that can segment a tongue with a clear counter. Patented instruments and algorithm are used in this study.It is recommended to use professional instruments and master computer technology. To begin, prepare the self-developed handheld lingual face diagnostic instrument to collect lingual face images of patients. Fill in the patient's name, gender, age, and disease on the computer page.Next, ask the patient to sit upright and position the image acquisition device connected to the computer. Then, position the face of the patient in the image acquisition instrument and ask them to extend their tongue out of their mouth to the maximum extent. Verify through the images on the computer screen that the patient is in the correct position and that the tongue and face are fully exposed.Press the shoot button on the computer screen three times to capture three images. Then, manually select the collected images of the face and the tongue. Filter them according to the Image Acquisition page of the software.Collect three images from each patient at a time and export the images to a folder for manual screening. Next, select a standard, fully exposed, well-illuminated, and clear images, the sample for algorithm training and testing. Delete the images that do not meet the criteria of a standard image.The criteria for exclusion are incomplete tongue and face exposure and images that are too dark. To perform tongue image segmentation, open LabelMe and click on the Open button in the upper left corner of the interface. Select the folder containing the image to be segmented and open the photos.Click the Create Polygon button to track the points. Track the tongue and lingual shapes, name them according to the selected areas, and save. On finishing all tracking, click Save to save the image to the data folder.Set the device to capture images with a size of 1080x1920 pixels and ensure the size of the filled image is 1920x1920 pixels. Manually screen the the images of each patient to select and retain one from the three images that were taken for uncontrollable factors, such as subject blinking and lens blocking. For training the model, collect data from 200 individuals or 600 images and retain about 200 images that are usable after the screening.Randomly divide all the tongue images according to the image number, placing 70%of them into the Training Set and 30%of the images into the Test Set folder. Download and install Anaconda Python and LabelMe from their official websites. Activate the environment and complete the installation and adjustment of the overall environment.Choose the appropriate model for the research purpose. In this study, UNet, SegNet, DeepLab Version 3, and PSPNet were selected for validation. Next, build the deep learning algorithm model in the installed environment.Tune the parameters by adjusting the values and complete the model training using the Training Set. Using LabelMe annotation and uniform image size methods, construct the required data set in conjunction with the research content. Examine the results of the segmentation and evaluate the model performance based on four metrics, precision, recall, mean pixel accuracy or MPA, and MIOU, to get a more comprehensive evaluation.Compare the generated values of the four models. The higher the value, the higher the segmentation accuracy and the better the performance of the model. According to the index results, the UNet algorithm is superior to the other algorithms in MIOU, MPA, precision, and recall, and its segmentation accuracy is also higher.PSPNet is better than DeepLab Version 3 in MIOU, MPA, and recall, while the DeepLab Version 3 model is lower than the segment model in all indexes. In tongue segmentation, the outer red area in the image results from tongue segmentation and the inner green area results from tongue coating segmentation. In this study, the unit algorithm performed well in tongue segmentation, and further research can be conducted based on the unit algorithms.

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2.0K vues.  Chengdu University of Traditional Chinese Medicine. Ce protocole utilise un algorithme d’apprentissage profond pour segmenter les images de la langue et sépare automatiquement la langue de l’image entière, qui peut être utilisée pour des recherches ultérieures. Ce protocole compare les performances de l’algorithme de segmentation d’image à quatre langues pour sélectionner l’algorithme de base qui peut segmenter une langue avec un compteur clair. Des instruments et des algorithmes brevetés sont utilisés dans cette étude....

Vidéo: Objectification of Tongue Diagnosis in Traditional Medicine, Data Analysis, and Study Application

The present study employed U-Net and other deep learning algorithms to segment a tongue image and compared the segmentation results to investigate the objectification of tongue...

gavage strategy for decoction formula of traditional chinese medicine in osteosarcoma model mice

Author Spotlight: Standardized Herbal Decoction Protocol for Enhanced Animal Studies

The protocol provides a detailed standardization process of decoction formula and gavage technique with Yiqi Jiedu decoction in the osteosarcoma mouse model as an example. It describes animal protection and improves reliability of research data, providing effective strategies for investigating therapeutic efficacy and molecular mechanisms of decoction formulas in...

Gavage Strategy for Decoction Formula of Traditional Chinese Medicine in Osteosarcoma Model Mice

enema of traditional chinese medicine for patients with severe acute pancreatitis

Enema of Traditional Chinese Medicine for Patients with Severe Acute Pancreatitis

An enema is a type of traditional Chinese medicine (TCM) therapy used to treat severe acute pancreatitis (SAP). The therapy involves instilling a TCM liquid into the rectum and colon through the anus to keep the liquid in the intestine, absorbing through the intestinal mucosa. This study aims to standardize the enema procedure for SAP...

acupoint catgut embedding therapy in traditional chinese medicine for managing allergic rhinitis

Author Spotlight: Exploring the Therapeutic Effect of Acupoint Catgut Embedding in Allergic Rhinitis 

This protocol details the procedure for acupoint burrowing and demonstrates its effectiveness in treating allergic...

Acupoint Catgut Embedding Therapy in Traditional Chinese Medicine for Managing Allergic Rhinitis

a traditional chinese medicine characteristic therapy for bronchial asthma: moxibustion

A Traditional Chinese Medicine Characteristic Therapy for Bronchial Asthma: Moxibustion

This protocol presents a curative procedure of moxibustion in treating patients with bronchial...

fabrication of tongue extracellular matrix and reconstitution of tongue squamous cell carcinoma in vitro

Fabrication of Tongue Extracellular Matrix and Reconstitution of Tongue Squamous Cell Carcinoma In Vitro

A method is shown here for the preparation of the tongue extracellular matrix (TEM) with efficient decellularization. The TEM can be used as functional scaffolds for the reconstruction of a tongue squamous cell carcinoma (TSCC) model under static or stirred culture...

Fabrication of Tongue Extracellular Matrix and Reconstitution of Tongue Squamous Cell Carcinoma In Vitro

visualization of metabolites identified in the spatial metabolome of traditional chinese medicine using desi-msi

Visualization of Metabolites Identified in the Spatial Metabolome of Traditional Chinese Medicine Using DESI-MSI

In this study, a series of methods are presented to prepare DESI-MSI samples from plants, and a procedure of DESI assembly installation, MSI data acquisition, and processing is described in detail. This protocol can be applied in several conditions for acquiring spatial metabolome information in...

identifying disease targets and active components in traditional chinese medicine for anus eczema

Identifying Disease Targets and Active Components in Traditional Chinese Medicine for Anus Eczema

preparation and application of zhuang medicine threads for moxibustion in dysmenorrhea treatment

Preparation and Application of Zhuang Medicine Threads for Moxibustion in Dysmenorrhea Treatment

behavioral and network pharmacology-based analyses for the traditional mongolian medicine zadi-5 in a rat model of depression

Behavioral and Network Pharmacology-Based Analyses for the Traditional Mongolian Medicine Zadi-5 in a Rat Model of Depression

The present protocol describes a method for the behavioral test validation and bioinformatical prediction of the therapeutic efficacy of Zadi-5, a traditional Mongolian medicine, in...

formulation of a gavage decoction using traditional chinese medicine and its administration into mice

Formulation of a Gavage Decoction Using Traditional Chinese Medicine and its Administration into Mice

press needle: a traditional chinese medicine therapy for myopia patients with dry eye

Author Spotlight: Exploring Press Needle Efficacy and Underlying Molecular Pathways

Here, we present a protocol to standardize the operation of acupuncture in the treatment of myopia with dry eye.

Press Needle: A Traditional Chinese Medicine Therapy for Myopia Patients with Dry Eye

a computer-based platform for aiding clinicians in eating disorder analysis and diagnosis

A Computer-Based Platform for Aiding Clinicians in Eating Disorder Analysis and Diagnosis

Diagnosing eating disorders in healthcare is challenging. Therefore, the present protocol develops an algorithm based on 949 patient responses to a questionnaire, with the diagnosis displayed on an easy-to-use web-based interface. This system facilitates the accurate diagnosis of eating disorders while excluding those believed to have an eating disorder.

acupoint application combined with ear plaster therapy for treating sleep disorders with acute exacerbation of chronic obstructive pulmonary disease

Author Spotlight: Traditional Chinese Medicine for Sleep Disorders in Acute COPD &#8212; A Safe, Cost-Effective Approach

The present protocol describes the specific process and efficacy of the acupoint application combined with ear plaster therapy for sleep disorders in a patient with acute exacerbation of chronic obstructive pulmonary disease, including medical devices, selection of acupoints, operating methods, and posttreatment...

Acupoint and Ear Plaster Therapy for Treating Sleep Disorders in AECOPD Patients

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acupoint application combined with acupressure as an adjunctive therapy for chemotherapy-induced nausea and vomiting

Author Spotlight: Effective Non-Pharmacological Management of Chemotherapy-Induced Nausea and Vomiting Using Acupoint Application and Acupressure

Acupoint application combined with acupressure-assisted treatment has a good effect on chemotherapy-induced nausea and vomiting and is more convenient and...

Enhancing CINV Management: Traditional Chinese Medicine Approaches

medicated thread moxibustion therapy of zhuang medicine and its application in a rat model of cold-congealing syndrome of primary dysmenorrhea

Author Spotlight: Demonstrating Therapeutic Effect of Automated Zhuang Medicine Line Moxibustion Simulator

This article demonstrates the standardized operation process of Zhuang medicine line moxibustion in experimental animals. To achieve repeatability and accuracy of Zhuang medicine line moxibustion, we developed a simulator to maintain strength, heat, and frequency. We constructed a dysmenorrhea model for experimental verification in rats with cold coagulation...

Standardized Operation Process of Zhuang Medicine Line Moxibustion in Rats

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arl spectral fitting as an application to augment spectral data via franck-condon lineshape analysis and color analysis

ARL Spectral Fitting as an Application to Augment Spectral Data via Franck-Condon Lineshape Analysis and Color Analysis

This protocol introduces Franck-Condon Lineshape Analyses (FCLSA) of emission spectra and serves as a tutorial for the use of ARL Spectral Fitting software. The open-source software provides an easy and intuitive way to perform advanced analysis of emission spectra including excited state energy calculations, CIE color coordinate determination, and...

electronic tongue generating continuous recognition patterns for protein analysis

Electronic Tongue Generating Continuous Recognition Patterns for Protein Analysis

A novel approach is described for construction of electronic tongue (eT), which greatly simplifies the design and production of sensing materials, and allows the eT to generate continuous evolution profiles and landscapes for samples in liquid. The obtained eT is efficient for common protein analysis such as discrimination.

basics of multivariate analysis in neuroimaging data

Basics of Multivariate Analysis in Neuroimaging Data

The current article describes the basics of multivariate analysis and contrasts it to the more commonly used voxel-wise univariate analysis. Both types of analysis are applied to a clinical-neuroscience data set. Supplementary split-half simulations show better replication of the multivariate results in independent data...

hydra, a computer-based platform for aiding clinicians in cardiovascular analysis and diagnosis

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

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...

Technique to Collect Fungiform (Taste) Papillae from Human Tongue

The sense of taste is critical for human life. It informs the body about the quality of food that will be potentially ingested and stimulates metabolic processes that prepare the alimentary canal for digestion. Steady progress is being made towards understanding the early biochemical and molecular events underlying taste transduction (for a review, Breslin and Spector, 20081). However, progress to date has largely resulted from animal models. Yet, since marked differences in receptor specificity and receptor density vary among species, human taste transduction will only be understood by using human taste tissue. Here we describe a biopsy technique to collect human fungiform papillae, visible as rounded pink anterior structures, about 0.5 mm in diameter that contain taste buds. These biopsied papillae are used for several purposes including the isolation of viable taste bud cells, in situ hybridization, immunohistochemistry and, through techniques of molecular biology, the identification of taste-specific novel proteins.

Technique to Collect Fungiform Taste Papillae from Human Tongue

Knowledge of molecular mechanisms underlying gustatory transduction has recently enjoyed significant advances, largely due to using animal models. However, the wide diversity in taste sensitivity and specificity among mammals warrants studies in human tissue. We describe a biopsy technique to collect living taste cells from the papillae on human tongue.

The sense of taste is critical for human life. It informs the body about the quality of food that will be potentially ingested and stimulates metabolic ...

Microarray-based Identification of Individual HERV Loci Expression: Application to Biomarker Discovery in Prostate Cancer

The prostate-specific antigen (PSA) is the main diagnostic biomarker for prostate cancer in clinical use, but it lacks specificity and sensitivity, particularly in low dosage values1&#8203;&#8203;. &#8216;How to use PSA&#39; remains a current issue, either for diagnosis as a gray zone corresponding to a concentration in serum of 2.5-10 ng/ml which does not allow a clear differentiation to be made between cancer and noncancer2 or for patient follow-up as analysis of post-operative PSA kinetic parameters can pose considerable challenges for their practical application3,4. Alternatively, noncoding RNAs (ncRNAs) are emerging as key molecules in human cancer, with the potential to serve as novel markers of disease, e.g. PCA3 in prostate cancer5,6 and to reveal uncharacterized aspects of tumor biology. Moreover, data from the ENCODE project published in 2012 showed that different RNA types cover about 62% of the genome. It also appears that the amount of transcriptional regulatory motifs is at least 4.5x higher than the one corresponding to protein-coding exons. Thus, long terminal repeats (LTRs) of human endogenous retroviruses (HERVs) constitute a wide range of putative/candidate transcriptional regulatory sequences, as it is their primary function in infectious retroviruses. HERVs, which are spread throughout the human genome, originate from ancestral and independent infections within the germ line, followed by copy-paste propagation processes and leading to multicopy families occupying 8% of the human genome (note that exons span 2% of our genome). Some HERV loci still express proteins that have been associated with several pathologies including cancer7-10. We have designed a high-density microarray, in Affymetrix format, aiming to optimally characterize individual HERV loci expression, in order to better understand whether they can be active, if they drive ncRNA transcription or modulate coding gene expression. This tool has been applied in the prostate cancer field (Figure 1).

Human endogenous retroviruses (HERV), which occupy 8% of the human genome, retain scarce coding capacities but a hundred thousand long terminal repeats (LTRs). A custom Affymetrix microarray was designed to identify individual HERV locus expression and was used on prostate cancer tissues as a proof of concept for future clinical studies.

The prostate-specific antigen (PSA) is the main diagnostic biomarker for prostate cancer in clinical use, but it lacks specificity and sensitivity, ...

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.

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

High-Throughput Analysis of Optical Mapping Data Using ElectroMap

Optical mapping is an established technique for high spatio-temporal resolution study of cardiac electrophysiology in multi-cellular preparations. Here we present, in a step-by-step guide, the use of ElectroMap for analysis, quantification, and mapping of high-resolution voltage and calcium datasets acquired by optical mapping. ElectroMap analysis options cover a wide variety of key electrophysiological parameters, and the graphical user interface allows straightforward modification of pre-processing and parameter definitions, making ElectroMap applicable to a wide range of experimental models. We show how built-in pacing frequency detection and signal segmentation allows high-throughput analysis of entire experimental recordings, acute responses, and single beat-to-beat variability. Additionally, ElectroMap incorporates automated multi-beat averaging to improve signal quality of noisy datasets, and here we demonstrate how this feature can help elucidate electrophysiological changes that might otherwise go undetected when using single beat analysis. Custom modules are included within the software for detailed investigation of conduction, single file analysis, and alternans, as demonstrated here. This software platform can be used to enable and accelerate the processing, analysis, and mapping of complex cardiac electrophysiology.

This protocol describes the setup and use of ElectroMap, a MATLAB-based open-source software platform for analysis of cardiac optical mapping data. ElectroMap provides a versatile high-throughput tool for analysis of optical mapping voltage and calcium datasets across a wide range of cardiac experimental models.

Optical mapping is an established technique for high spatio-temporal resolution study of cardiac electrophysiology in multi-cellular preparations. Here we ...

Preparation of Naringenin Solution for In Vivo Application

The preparation of a compound (phytochemical) solution is an overlooked but critical step prior to its application in studies such as drug screening. The complete solubilization of the compound is necessary for its safe use and relatively stable results. Here, a protocol for preparing naringenin solution and its intraperitoneal administration in a high-fat diet and streptozotocin (STZ)-induced diabetic model is demonstrated as an example. A small amount of naringenin (3.52-6.69 mg) was used to test its solubilization in solvents, including ethanol, dimethylsulfoxide (DMSO), and DMSO plus Tween 80 reconstituted in physiological saline (PS), respectively. Complete solubilization of the compound is determined by observing the color of the solution, the presence of precipitates after centrifugation (2000 x g for 30 s), or allowing the solution to stand for 2 h at room temperature (RT). After obtaining a stable compound/phytochemical solution, the final concentration/amount of the compound required for in vivo studies can be prepared in a solvent-only (no PS) stock solution, and then diluted/mixed with PS as desired. The antidiabetic osteoporotic effects of naringenin in mice (intraperitoneal administration at 20 mg/kg b.w., 2 mg/mL) were assessed by measuring blood glucose, bone mass (micro-CT), and bone resorption rate (TRAP staining and ELISA). Researchers looking for detailed organic/phytochemical solution preparations will benefit from this technique.

Preparation of Naringenin Solution for In Vivo Application

Here, the protocol presents the preparation of naringenin solution for in vivo intraperitoneal administration. Naringenin is fully dissolved in a mixture of dimethylsulfoxide, Tween 80, and saline. The antidiabetic osteoporotic effects of naringenin were assessed by blood glucose testing, tartrate-resistant acid phosphatase staining, and enzyme-linked immunosorbent assay.

The preparation of a compound (phytochemical) solution is an overlooked but critical step prior to its application in studies such as drug screening. The ...

An In Vitro Dissolution Determination of Multi-Index Components in Tibetan Medicine Rhodiola Granules

The composition of the Tibetan medicine Rhodiola granules (RG) is complex, and the overall quality of RG is difficult to determine. Therefore, establishing a method to determine the multi-component in vitro dissolution of RG is of great significance for quality control. This study uses the second paddle method of the fourth general rule 0931 from the Chinese Pharmacopoeia (2020 edition), compliant with apparatus 2 of the United States Pharmacopeia (USP). The dissolution apparatus was set to a rotation speed of 100 rpm with ultrapure water as the dissolution medium. A sample volume of 1 mL was collected at each timepoint. Furthermore, the cumulative dissolution of gallic acid, salidroside, and ethyl gallic acid in RG at different time points was determined by high-performance liquid chromatography (HPLC). Finally, the dissolution curves were drawn, and the curves were fitted to the GompertzMod, the Gompertz, the Logistic, and the Weibull equations. The results showed that the cumulative dissolution of gallic acid in RG was over 80% at 1 min, the cumulative dissolution of salidroside and ethyl gallic acid was over 65% at 5 min, and the cumulative dissolution of each index component decreased after 30 min. The curve fitting demonstrated that the GompertzMod equation was the best-fitting model for each index component of RG. In conclusion, the dissolution test method described in this protocol is simple, accurate, and reliable. It can characterize the dissolution behavior of the index components in RG in vitro, which provides a methodological reference for quality control of RG and quality evaluation of other ethnic compounds.

An In Vitro Dissolution Determination of Multi-Index Components in Tibetan Medicine Rhodiola Granules

Here, we test the dissolution of Rhodiola granules (RG) in vitro, draw dissolution curves of salidroside, gallic acid, and ethyl gallate in ultrapure water, and fit the curves to different mathematical models. This protocol provides information and guidance for in vivo bioequivalence and in vivo-in vitro correlation studies of RG.

The composition of the Tibetan medicine Rhodiola granules (RG) is complex, and the overall quality of RG is difficult to determine. Therefore, ...

The medicinal use of traditional Chinese medicine is mainly due to its secondary metabolites. Visualization of the distribution of these metabolites has become a crucial topic in plant science. Mass spectrometry imaging can extract huge volumes of data and provide spatial distribution information about these by analyzing tissue slices. With the advantage of high throughput and higher accuracy, desorption electrospray ionization mass spectrometry imaging (DESI-MSI) is often used in biological research and in the study of traditional Chinese medicine. However, the procedures used in this research are complicated and not affordable. In this study, we optimized sectioning and DESI imaging procedures and developed a more cost-effective method to identify the distribution of metabolites and categorize these compounds in plant tissues, with a special focus on traditional Chinese medicines. The study will promote the utilization of DESI in metabolite analysis and standardization of traditional Chinese medicine/ethnic medicine for research-related technologies.

In this study, a series of methods are presented to prepare DESI-MSI samples from plants, and a procedure of DESI assembly installation, MSI data acquisition, and processing is described in detail. This protocol can be applied in several conditions for acquiring spatial metabolome information in plants.

The medicinal use of traditional Chinese medicine is mainly due to its secondary metabolites. Visualization of the distribution of these metabolites has ...

Standardized Tuina Manipulation for Knee Osteoarthritis in Rats

Tuina Manipulation to Reduce Inflammation and Cartilage Loss in Knee Osteoarthritis Rats

Clinical trials suggest that Tuina manipulation is effective in treating knee osteoarthritis (KOA), while further studies are required to discover its mechanism. Therefore, the manipulation of animal models of knee osteoarthritis is critical. This protocol provides a standard process for Tuina manipulation on KOA rats and a preliminary exploration of the mechanism of Tuina for KOA. The press and kneading manipulation method (a kind of Tuina manipulation that refers to pressing and kneading the specific area of the body surface) is applied on 5 acupoints around the knee joint of rats. The force and frequency of the manipulation were standardized by finger pressure recordings, and the position of the rat during manipulation is described in detail in the protocol. The effect of manipulation can be measured by pain behavior tests and microscopic findings in synovial and cartilage. KOA rats showed significant improvement in pain behavior. The synovial tissue inflammatory infiltration was reduced in the Tuina group, and the expression of tumor necrosis factor (TNF)-&#945; was significantly lower. Compared to the control group, chondrocyte apoptosis was less in the Tuina group. This study provides a standardized protocol for Tuina manipulation on KOA rats and preliminary proof that the therapeutic effects of Tuina may be related to reducing synovial inflammation and delayed chondrocyte apoptosis.

Author Spotlight: Understanding the Mechanism of Tuina Manipulation in Rats 

We present a protocol for Tuina manipulation performed on plaster-immobilized-induced knee osteoarthritis rats. Based on the preliminary results, it suggested that the efficacy of the method relied on the reduction of inflammation and cartilage loss.

Clinical trials suggest that Tuina manipulation is effective in treating knee osteoarthritis (KOA), while further studies are required to discover its ...

Establishment of the KOA Model in Rabbits Using the Modified Videman Method

Application of Acupotomy in a Knee Osteoarthritis Model in Rabbit

Knee osteoarthritis (KOA) is one of the most frequently encountered diseases in the orthopedic department, which seriously reduces the quality of life of people with KOA. Among several pathogenic factors, the biomechanical imbalance of the knee joint is one of the main causes of KOA. Acupotomology believes that restoring the mechanical balance of the knee joint is the key to treating KOA. Clinical studies have shown that acupotomy can effectively reduce pain and improve knee mobility by reducing adhesion, contracture of soft tissues, and stress concentration points in muscles and tendons around the knee joint. 
In this protocol, we used the modified Videman method to establish a KOA model by immobilizing the left hindlimb in a straight position. We have outlined the method of operation and the precautions related to acupotomy in detail and evaluated the efficacy of acupotomy in conjunction with the theory of "Modulating Muscles and Tendons to Treat Bone Disorders" through the detection of the mechanical properties of quadriceps femoris and tendon, as well as cartilage mechanics and morphology. The results show that acupotomy has a protective effect on cartilage by adjusting the mechanical properties of the soft tissues around the knee joint, improving the cartilage stress environment, and delaying cartilage degeneration.

Author Spotlight: Investigating the Mechanism of Action of Acupotomy in Treating Knee Osteoarthritis

In this protocol, a knee osteoarthritis model was prepared using the modified Videman method, and the operation procedures and precautions of acupotomy are detailed. The effectiveness of acupotomy has been demonstrated by testing the mechanical properties of quadriceps femoris and tendon and the mechanical and morphological properties of cartilage.

Knee osteoarthritis (KOA) is one of the most frequently encountered diseases in the orthopedic department, which seriously reduces the quality of life of ...

&lt;i&gt;Desmodium caudatum &lt;/i&gt; Decoction&#039;s Impact on Intestinal Microbiota of Rats

Effects of Desmodium caudatum on Gastrointestinal Hormones and Intestinal Flora in Rats with Gastritis

In order to preliminarily explore the effects of Desmodium caudatum on gastritis and intestinal flora in rats, a chronic gastritis rat model was established using the classic sodium salicylate method. Eighteen SPF rats were divided into three groups: the control group (Group C), the model group (Group M), and the treatment group (Group T). Pathological sections of the gastric wall were taken from rats in each group. Furthermore, the concentrations of gastrin and malondialdehyde in the serum of rats in each group were determined by ELISA. Additionally, the effects of D. caudatum on the intestinal flora of rats with gastritis were explored through a detailed comparison of gut bacterial communities in the three groups, employing Illumina-based 16S rRNA gene sequencing. The results indicated that D. caudatum decoction could reduce the malondialdehyde content and increase the gastrin content. Moreover, D. caudatum decoction was found to enhance the diversity and abundance of intestinal flora, exerting a positive impact on the treatment of gastritis by regulating and restoring the intestinal flora.

Author Spotlight: Exploring the Mysteries of Sichuan&#039;s Herbal Medicine in Chinese Medicine Research

The present protocol describes the method for analyzing intestinal flora using Illumina-based 16S rRNA gene sequencing and provides a framework for evaluating the effectiveness of herbal decoctions.