We are grateful for the instrumentation and laboratory provided by the Mongolian medical faculty of the Inner Mongolian Medical University, China. This study was supported by the National Natural Sciences Foundation of China (81760762) and the Science and Technology Plan Project of the Health Commission of Inner Mongolia, China (202201300).

All the experimental protocols were approved by the Ethics of Animal Experiment Care Committee of Inner Mongolia Medical University and followed the guidelines of the National Institutes of Health on animal care and ethics. Male Sprague Dawley (SD) rats aged 8 weeks old (200 g &#177; 20 g) were housed in a room with a controlled temperature (22 &#176;C &#177; 2 &#176;C) and humidity (55% &#177; 15%) under a 12 h/12 h regulated light/dark cycle for 1 week. See Figure 1 for the workflow of the network pharmacology analysis.
1. Behavioral test in rats
<ol>
	<li>Establish a CUMS rat model
	<ol>
		<li>Apply the following stimuli combined with isolation for 28 days to all the rats, except for the controls: inversion of the light/dark cycle for 24 h, food deprivation for 24 h, water deprivation for 24 h, high-speed level shaking for 15 min (one time/s), tail clamp for 2 min, swimming in cold water (4 &#176;C) for 5 min, 45 &#176;C heat stimulation, and wet padding for 24 h (Table 1). Raise the rats in individual cages. 
		NOTE: Avoid repeating the same type of stimuli for consecutive days.&#160;&#160;The above procedures to establish a CUMS rat model have been approved by the animal ethics committee and described previously15.</li>
	</ol>
	</li>
	<li>Drug preparation
	<ol>
		<li>Pulverize the Zadi-5 pill in a grinder, and prepare a 1.16 g/mL solution in distilled water. Separately prepare a fluoxetine solution of 0.36 mg/mL in distilled water.</li>
	</ol>
	</li>
	<li>Drug administration
	<ol>
		<li>Divide the rats randomly into six groups (n = 10): control (CON), model (MOD), Zadi-5 group (Zadi-5, 1.6 g of Zadi-5/kg16), fluoxetine group (Fluoxetine, 3.6 mg of fluoxetine/kg). One time per day for 28 days, administer 1 mL/g per rat of the appropriate drug solution by gavage and treat the CON and MOD groups with an equal volume of distilled water. 
		NOTE: Gavage starts at the beginning of model establishment for all the groups.</li>
	</ol>
	</li>
	<li>Open-field test (OFT)
	<ol>
		<li>Divide a black box (50 cm x 50 cm x 30 cm) into nine square regions of equal area. Equip the box with a video tracking analysis system. One day after the last gavage, place the rat in the center square, and record its horizontal and vertical activities for 3 min.</li>
		<li>Score the number of squares crossed with all paws as a horizontal activity, and score standing and grooming as a vertical activity. After each test, clean the box with 75% alcohol to remove the smell of the rat for subsequent tests17.</li>
	</ol>
	</li>
	<li>Sucrose consumption test (SCT)
	<ol>
		<li>Weigh the respective bottles before and after consumption, and calculate the 60 min sucrose preference rates on day 0, day 7, day 14, day 21, and day 28 using equation (1): 
		&#8203;Sucrose consumption = <img alt="Equation 1" src="/files/ftp_upload/64832/64832eq01.jpg" style="margin: auto;" /> &#215; 100%&#160; &#160; &#160;(1)</li>
	</ol>
	</li>
	<li>Morris water maze (MWM)
	<ol>
		<li>Divide the pool into four quadrants. Order the quadrants from one to four, and place the hidden platform in the third quadrant, 1 cm below the water surface.</li>
		<li>Place the rat subject into the maze in different quadrants to look for the platform for 120 s, and record the latency time using the MWM video trail analysis system.</li>
		<li>Place the rat subject in a fixed position in the pool. If the subject cannot find the hidden platform in 120 s, record the latency as 120 s.</li>
		<li>Next, dislodge the platform, place the rat in the water, and record the number of zone-crossings for 120 s.</li>
		<li>Add milk to the pool for some level of opacity. Maintain the water temperature at 23 &#176;C &#177; 1 &#176;C during the experiment.</li>
	</ol>
	</li>
</ol>
2. Network pharmacological prediction
<ol>
	<li>Screen the active components in Zadi-5.
	<ol>
		<li>Browse the Traditional Chinese Medicine Systems Pharmacology (TCMSP, https://old.tcmsp-e.com/tcmsp.php), and input&#160;&#34;Myristicae Semen Seeds,&#34; &#34;Aucklandiae Radix roots,&#34; and &#34;Piperis Longi Fructus&#34; in the &#34;herb name&#34; section to obtain the names of chemicals. Set the pharmacokinetic index of oral bioavailability (OB) to be &#62;30% and the drug-like (DL) index to be &#62;0.18 (Supplementary File 1).</li>
		<li>Search &#34;Rou Dou Kou&#34; (Myristica fragrans Houtt), &#34;Tu Mu Xiang&#34; (Inula helenium L.), &#34;Mu Xiang&#34; (Aucklandia lappa Decne.), &#34;Guang Zao&#34; (Choerospondias axillaris Roxb. Burtt Hill), and &#34;Bi Ba&#34; (Piper longum L.) in the Chinese Medicine Pharmacopeia (http://www.zhongyaocai360.com/zhongguoyaodian/) to identify the chemical names of each component.</li>
		<li>Search the identified chemical names in PubChem (https://pubchem.ncbi.nlm.nih.gov/) to find the isomeric SMILES or InChIkey</li>
	</ol>
	</li>
	<li>Identify the target proteins of the active components in Zadi-5.
	<ol>
		<li>Identify the target proteins of the active components using SEA (http://sea.bkslab.org/), BATMAN (http://bionet.ncpsb.org.cn/batman-tcm/), and Swiss Target Prediction (http://www.swisstargetprediction.ch/) with isomeric SMILES or InChIkey, and find the overlapping proteins.</li>
		<li>Use the protein database UniProt (http://www.uniprot.org/uploadlists/) to convert the identified targets to unified gene names.</li>
	</ol>
	</li>
	<li>Search for the target proteins for depression.
	<ol>
		<li>Search and identify the potential protein targets for depression by using the keywords &#34;depression&#34; and &#34;depressive disorder&#34; in Genecards (https://www.genecards.org/), Disgenet (https://www.disgenet.org/), and Drugbank (https://www.drugbank.com/).</li>
	</ol>
	</li>
	<li>Find the target genes.
	<ol>
		<li>Browse the Venn diagram (http://bioinformatics.psb.ugent.be/webtools/Venn/), upload the targets of the active components of Zadi-5 in List-1, upload the targets for depression in List-2, and submit. Obtain the Venn diagram, and filter out the overlapping target candidates.</li>
	</ol>
	</li>
	<li>Construct the network.
	<ol>
		<li>Construct a spreadsheet called &#34;Type and Network&#34; (Supplementary File 2). &#34;Type&#34; is the network&#39;s signature, and &#34;Network&#34; illustrates the relationship between the signs.</li>
		<li>Export the &#34;Type and Network&#34; to Cytoscape v3.9.0 to construct the network &#34;Zadi-5 herbs-ingredients-disease targets.&#34;</li>
	</ol>
	</li>
	<li>Analyze the protein-protein interaction (PPI) network of the target candidates.
	<ol>
		<li>Set the common targets in the STRING database (https://cn.string- db.org/) to analyze their interactions. Set the&#160;protein type as &#34;homo sapiens.&#34; Set the interaction threshold value to 0.9, and select only the experimentally verified types. Do not display the lonely island nodes.</li>
	</ol>
	</li>
	<li>Conduct a Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) enrichment analyses of the target-related pathways.
	<ol>
		<li>Paste 86 potential antidepressant targets of Zadi-5 into the start analysis bracket in DAVID (https:// david.ncifcrf.gov/) to study the related signaling pathways by performing the Gene Ontology (GO) function-including biological process, cellular component, and molecular function-and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses. 
		NOTE: KEGG is visualized in the bubble chart using an online IMageGP (http://www.ehbio.com /ImageGP/index.php/Home/Index/). The bubble size represents the number of targets enriched in the indicated pathway, and the bubble color represents the enrichment&#8217;s&#160;&#8203;P-value.</li>
	</ol>
	</li>
	<li>Construct the network to illustrate the active compounds in Zadi-5 that interact with the PI3K-AKT signaling pathway.
	<ol>
		<li>Download the KEGG pathway document, select the genes of the PI3K-AKT pathway from the enrichment analysis, and paste them on the spreadsheet to construct a &#34;Type and Network&#34; document.</li>
		<li>Export the &#34;Type and Network&#34; document to the Cytoscape to generate the &#34;PI3K-AKT visualized compounds-targets-pathways network&#34; (Supplementary File 3). 
		&#8203;NOTE: &#34;Type&#34; is the network&#39;s signature, and &#34;Network&#34; illustrates the relationship between the signs.</li>
	</ol>
	</li>
</ol>
3. Statistical analysis
<ol>
	<li>Use a one-way analysis of variance (ANOVA), followed by Duncan&#39;s post hoc test, to determine the significant differences in biochemical and gene expression parameters. Calculate the mean &#177; standard deviation (SD), and visualize the data. Consider P &#60; 0.05 as statistically significant.</li>
</ol>

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The authors have no conflicts of interest to disclose.

Depression is a mental disease characterized by low mood, anhedonia, and a lack of energy. This disorder is accompanied by distraction, cognitive dysfunction, social withdrawal, insomnia, sexual dysfunction, and gastrointestinal diseases18,19. In the study of depression, establishing an animal model is crucial for understanding the pathological mechanisms and effects of new drugs. In this study, a CUMS-induced rat depression model was established through the irri...

Depression, also known as major depressive disorder (MDD), is a severe neuropsychiatric disease responsible for growing medical and economic burdens on society. Due to the associated complexity, morbidity, and mortality rates, a significant amount of research has been conducted to find remedies for the disorder1,2. According to a mental health survey by the World Health Organization, around 350 million people currently suffer from depression and its associated symptoms worldwide. It is predicted that depression will overtake cancer and cardiovascular diseases as the leading cause of disease burden globally by 2030. Thus, the prevention and treatment of depression will become a global priority in the near future3. The pathogenesis of MDD has not yet been elucidated. Still, it is commonly attributed to the following factors: genetic predisposition, dysfunction of the hypothalamus-pituitary-adrenal axis, reductions in neurotransmitter secretion, neuroimmune dysregulation-induced neuroinflammation, cell apoptosis, and reduced cell proliferation4,5. 
Among these factors, neuroimmune dysregulation-induced neuroinflammation and altered secretion of neurotrophic factors have received particular attention for their roles in the development of depression and many other psychiatric diseases6. In the past decade, scholars have demonstrated that the hippocampus is the dominant site for regenerative nerve functions and is involved in regulating emotion and cognition. In this regard, the hippocampal neurons are recognized as novel therapeutic targets for antidepressant medicines under development7,8. Moreover, the hippocampus is also reported to be involved in short-term and long-term memory in learning and consolidating memories. Specifically, the shortage of pyramidal neurons in the CA1 region of the hippocampus causes retrograde and anterograde amnesia9. A typical antidepressant therapeutic strategy aims to enhance cell proliferation and neurogenesis in the dentate gyrus of the hippocampus. Natural product-derived compounds and small molecules synthesized based on medicinal chemistry techniques are considered the primary sources of innovative therapeutic agents for various neuropsychiatric conditions. 
Traditional Mongolian medicines, which have a long history and a well-supported theoretical medical system, have descended from the nomads of the Mongolian plateau These medicines display multi-target and multi-pathway effects due to the various medicinal components that act in concert to generate synergistic functions. Zadi-5 is a well-established formulation among such drugs and was first recorded in "Clinical Experience of Dr. Gao Shi," written by an outstanding Mongolian clinician called Dr. Gao Shi (1804-1876). It has been clinical practice for a long time in Mongolia to use these pills to treat the symptoms of distress, palpitation, irritation, and cardiac stabbing pain10,11. Moreover, Zadi-5 has proven effects on alleviating post-stroke depression in affected patients12. The recent experimental research on CUMS has revealed that the Zadi-5 formulation alleviates depression by regulating the central neurotransmitters13; indeed, with Zadi-5, increased levels of brain-derived neurotrophic factor (BDNF) and tyrosine kinase receptor B (TrkB) have been detected and correlated with improved learning and memory in a rat model of depression14. However, the exact mechanism of action of Zadi-5 for such alleviation of depression has not been elucidated.
This study aimed to demonstrate the therapeutic effects of Zadi-5 against depression in rats using a behavioral test and identify the components of Zadi-5 using Traditional Chinese Medicine Systems Pharmacology (TCMSP) and Swiss Target Prediction to predict the potential mechanisms underlying the efficacy of Zadi-5, a traditional Mongolian medicine, in treating depression.

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			<td>Cytoscape software</td>
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			<td>&#160;version 3.7.0</td>
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			<td>Fluoxetine</td>
			<td>Lilly Suzhou Pharmaceutical Co., Ltd</td>
			<td>J20160029</td>
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			<td>Morris water maze video trail analysing system&#160;</td>
			<td>Tai Meng Tech Co., Ltd</td>
			<td>WMT-200</td>
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			<td>Sprague Dawley rats</td>
			<td>Beijing Biotechnology Co., Ltd, China</td>
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			<td>Tai Meng Tech Co., Ltd</td>
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			<td>Zadi-5 pill</td>
			<td>Pharmaceutical Preparation Center of International Mongolian Hospital, Inner Mongolia, China</td>
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behavioral and network pharmacology-based analyses for the traditional mongolian medicine zadi-5 in a rat model of depression

Zadi-5 is a traditional Mongolian medicine that is widely used for the treatment of depression and symptoms of irritation. Although the therapeutic effects of Zadi-5 against depression have been indicated in previously reported clinical studies, the identity and impact of the active pharmaceutical compounds present in the drug have not been fully elucidated.&#160;This study used network pharmacology to predict the drug composition and identify the therapeutically active compounds in Zadi-5 pills. Here, we established a rat model of chronic unpredicted mild stress (CUMS) and conducted an open field test (OFT), Morris water maze (MWM) analysis, and sucrose consumption test (SCT) to investigate the potential therapeutic efficacy of Zadi-5 in depression. This study aimed to demonstrate Zadi-5&#39;s therapeutic effects for depression and predict the critical pathway of the action of Zadi-5 against the disorder. The vertical and horizontal scores (OFT), SCT, and zone crossing numbers of the fluoxetine (positive control) and Zadi-5 groups were significantly higher (P &#60; 0.05) than those of the CUMS group rats without treatment. According to the results of network pharmacology analysis, the PI3K-AKT pathway was found to be essential for the antidepressant effect of Zadi-5.

Behavioral test in animals 
Results of the behavioral tests in the CUMS-induced rat depression model 
No significant differences between the tested groups were found for the OFT score, sucrose consumption, and MWM analysis before CUMS stimulation. After establishing the CUMS model, the MOD group&#39;s vertical and horizontal scores were lower than those of the CON group (P &#60; 0.05). Compared with the MOD group, the vertical and horizontal scores of the POS and Zadi-5 groups ...

Watch this Scientific Journal Video about Behavioral and Network Pharmacology-Based Analyses for the Traditional Mongolian Medicine Zadi-5 in a Rat Model of Depression at JoVE.com

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 depression.

Zadi-5 is a traditional Mongolian medicine that is widely used for the treatment of depression and symptoms of irritation. Although the therapeutic ...

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405 Views.  Inner Mongolia Medical University. 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 depression.

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

Biomarkers in an Animal Model for Revealing Neural, Hematologic, and Behavioral Correlates of PTSD

Identification of biomarkers representing the evolution of the pathophysiology of Post Traumatic Stress Disorder (PTSD) is vitally important, not only for objective diagnosis but also for the evaluation of therapeutic efficacy and resilience to trauma. Ongoing research is directed at identifying molecular biomarkers for PTSD, including traumatic stress induced proteins, transcriptomes, genomic variances and genetic modulators, using biologic samples from subjects' blood, saliva, urine, and postmortem brain tissues. However, the correlation of these biomarker molecules in peripheral or postmortem samples to altered brain functions associated with psychiatric symptoms in PTSD remains unresolved. Here, we present an animal model of PTSD in which both peripheral blood and central brain biomarkers, as well as behavioral phenotype, can be collected and measured, thus providing the needed correlation of the central biomarkers of PTSD, which are mechanistic and pathognomonic but cannot be collected from people, with the peripheral biomarkers and behavioral phenotypes, which can.
Our animal model of PTSD employs restraint and tail shocks repeated for three continuous days - the inescapable tail-shock model (ITS) in rats. This ITS model mimics the pathophysiology of PTSD 17, 7, 4, 10. We and others have verified that the ITS model induces behavioral and neurobiological alterations similar to those found in PTSD subjects 17, 7, 10, 9. Specifically, these stressed rats exhibit (1) a delayed and exaggerated startle response appearing several days after stressor cessation, which given the compressed time scale of the rat's life compared to a humans, corresponds to the one to three months delay of symptoms in PTSD patients (DSM-IV-TR PTSD Criterian D/E 13), (2) enhanced plasma corticosterone (CORT) for several days, indicating compromise of the hypothalamopituitary axis (HPA), and (3) retarded body weight gain after stressor cessation, indicating dysfunction of metabolic regulation.
The experimental paradigms employed for this model are: (1) a learned helplessness paradigm in the rat assayed by measurement of acoustic startle response (ASR) and a charting of body mass; (2) microdissection of the rat brain into regions and nuclei; (3) enzyme-linked immunosorbent assay (ELISA) for blood levels of CORT; (4) a gene expression microarray plus related bioinformatics tools 18. This microarray, dubbed rMNChip, focuses on mitochondrial and mitochondria-related nuclear genes in the rat so as to specifically address the neuronal bioenergetics hypothesized to be involved in PTSD.

We describe a rat model of post traumatic stress disorder (PTSD) that reveals the persistent alterations in neuroendocrine function and the delayed long-term, exaggerated fear response, characteristic of PTSD patients. The animal model and methods described here are useful for correlating biomarkers in brain nuclei, which are mechanistic but cannot be measured in patients, with biomarkers in peripheral white blood cells, which can.

Identification of biomarkers representing the evolution of the pathophysiology of Post Traumatic Stress Disorder (PTSD) is vitally important, not only for ...

Preparation of Herbal Medicine: Er-Xian Decoction and Er-Xian-containing Serum for In Vivo and In Vitro Experiments

Traditional herbal medicine, an alternative medicine in the clinical setting, has received increased attention in recent years. Before delivery to the body, an additional extraction procedure is commonly required to release the active constituents from raw herbs. Water decoction is a classical extraction procedure that is still broadly used in the clinical settings. Here, we propose a detailed protocol for er-xian decoction (EXD) in order to apply herbal decoctions to experimental studies. The calculation of an animal-appropriate dose is described, as well as the four main steps of EXD: soaking, water decoction, filtration, and concentration. In addition, serum-containing EXD is introduced to rats as a means of in vitro validation. Here, rats were orally administered EXD for three days. Blood samples were then collected, inactivated, centrifuged, and filtered. The serum, diluted with the culture medium, can be utilized to treat cells or tissues in vitro. For example, EXD was applied to both in vivo and in vitro studies and demonstrated that EXD enhances osteogenesis. This protocol can be used as a reference for the preparation and application of herbal medicines.

Preparation of Herbal Medicine: Er-Xian Decoction and Er-Xian-containing Serum for In Vivo and In Vitro Experiments

Here, we present the preparation of an er-xian decoction (EXD) in four steps&#8212;soaking, decoction, filtration, and concentration&#8212;and demonstrate the administration of a prepared EXD-containing serum to rats. These methods are applicable to the in vivo and in vitro study of herbal decoctions such as traditional Chinese medicines.

Traditional herbal medicine, an alternative medicine in the clinical setting, has received increased attention in recent years. Before delivery to the ...

Reduced Complications after Arterial Reconnection in a Rat Model of Orthotopic Liver Transplantation

The rat orthotopic liver transplantation (OLT) model is a powerful tool to study acute and chronic rejection. However, it is not a complete representation of human liver transplantation due to the absence of arterial reconnection. Described here is a modified transplantation procedure that includes the incorporation of hepatic artery (HA) reconnection, leading to a marked improvement in transplant outcomes. With a mean anhepatic time of 12 min and 14 s, HA reconnection results in improved perfusion of the transplanted liver and an increase in long-term recipient survival from 37.5% to 88.2%. This protocol includes the use of 3D-printed cuffs and holders to connect the portal vein and infrahepatic inferior vena cava. It can be implemented for studying multiple aspects of liver transplantation, from immune response and infection to technical aspects of the procedure. By incorporating a simple and practical method for arterial reconnection using a microvascular technique, this modified rat OLT protocol closely mimics aspects of human liver transplantation and will serve as a valuable and clinically relevant research model.

The goal of this study is to modify the rat orthotopic liver transplant model to better represent human liver transplantation and improve recipient survival. The presented method reestablishes hepatic arterial inflow by connecting the donor liver&#39;s common hepatic artery to the recipient liver&#39;s proper hepatic artery.

The rat orthotopic liver transplantation (OLT) model is a powerful tool to study acute and chronic rejection. However, it is not a complete representation ...

Acupuncture in a Rat Model of Asthma

A high platform can fix rats without restriction and completely expose the acupoints on the back during acupuncture manipulation. This article describes methods for the fabrication of the high platform, establishes a rat model of asthma and measures changes in respiratory function using a noninvasive and real-time whole-body plethysmography (WBP) system.

A high platform can fix rats without restriction and completely expose the acupoints on the back during acupuncture manipulation. This article describes ...

A Protocol for Constructing a Rat Wound Model of Type 1 Diabetes

A single high dose of streptozotocin injection followed by full-thickness skin excision on the dorsum of rats is a common method for constructing animal models of type 1 diabetic wounds. However, improper manipulation can lead to model instability and high mortality in rats. Unfortunately, there are few existing guidelines on type 1 diabetic wound modeling, and they lack detail and do not present specific reference strategies. Therefore, this protocol details the complete procedure for constructing a type 1 diabetic wound model and analyzes the progression and angiogenic characteristics of the diabetic wounds. Type 1 diabetic wound modeling involves the following steps: preparation of the streptozotocin injection, induction of type 1 diabetes mellitus, and construction of the wound model. The wound area was measured on day 7 and day 14 after wounding, and the skin tissues of the rats were extracted for histopathological and immunofluorescence analysis. The results revealed that type 1 diabetes mellitus induced by 55 mg/kg streptozotocin was associated with lower mortality and a high success rate. The blood glucose levels were relatively stable after 5 weeks of induction. The diabetic wound healing rate was significantly lower than that of normal wounds on day 7 and day 14 (p &#60; 0.05), but both could reach more than 90% on day 14. Compared with the normal group, the epidermal layer closure of diabetic wounds on day 14 was incomplete and had delayed re-epithelialization and significantly lower angiogenesis (p &#60; 0.01). The type 1 diabetic wound model constructed based on this protocol has the characteristics of chronic wound healing, including poor closure, delayed re-epithelialization, and decreased angiogenesis compared to normal rat wounds.

The streptozotocin-induced diabetic wound model in male SD rats is currently the most widely used model for studying wound healing in type I diabetes mellitus. This protocol describes the methods used to construct this model. It also presents and addresses potential challenges and examines the progression and angiogenic characteristics of diabetic wounds.

A single high dose of streptozotocin injection followed by full-thickness skin excision on the dorsum of rats is a common method for constructing animal ...

Network Pharmacology Prediction and Metabolomics Validation of the Mechanism of Fructus Phyllanthi against Hyperlipidemia

Hyperlipidemia has become a leading risk factor for cardiovascular diseases and liver injury worldwide. Fructus Phyllanthi (FP) is an effective drug against hyperlipidemia in Traditional Chinese Medicine (TCM) and Indian Medicine theories, however the potential mechanism requires further exploration. The present research aims to reveal the mechanism of FP against hyperlipidemia based on an integrated strategy combining network pharmacology prediction with metabolomics validation. A high-fat diet (HFD)-induced mice model was established by evaluating the plasma lipid levels, including total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C). Network pharmacology was applied to find out the active ingredients of FP and potential targets against hyperlipidemia. Metabolomics of plasma and liver were performed to identify differential metabolites and their corresponding pathways among the normal group, model group, and intervention group. The relationship between network pharmacology and metabolomics was further constructed to obtain a comprehensive view of the process of FP against hyperlipidemia. The obtained key target proteins were verified by molecular docking. These results reflected that FP improved the plasma lipid levels and liver injury of hyperlipidemia induced by a HFD. Gallic acid, quercetin, and beta-sitosterol in FP were demonstrated as the key active compounds. A total of 16 and six potential differential metabolites in plasma and liver, respectively, were found to be involved in the therapeutic effects of FP against hyperlipidemia by metabolomics. Further, integration analysis indicated that the intervention effects were associated with CYP1A1, AChE, and MGAM, as well as the adjustment of L-kynurenine, corticosterone, acetylcholine, and raffinose, mainly involving tryptophan metabolism pathway. Molecular docking ensured that the above ingredients acting on hyperlipidemia-related protein targets played a key role in lowering lipids. In summary, this research provided a new possibility for preventing and treating hyperlipidemia.

The present protocol describes an integrated strategy for exploring the key targets and mechanisms of Fructus Phyllanthi against hyperlipidemia based on network pharmacology prediction and metabolomics verification.

Hyperlipidemia has become a leading risk factor for cardiovascular diseases and liver injury worldwide. Fructus Phyllanthi (FP) is an effective drug ...

A Traditional Chinese Medicine Characteristic Therapy for Bronchial Asthma: Moxibustion

Bronchial asthma is a chronic inflammatory disease of the airway which can lead to symptoms such as recurrent wheezing, shortness of breath, chest tightness, or cough, as a result of increased airway reactivity. With high diurnal variation, these symptoms often occur or worsen at night or in the morning. By burning and roasting the Chinese medica above human acupoints, moxibustion is a type of treatment that stimulates the activity of the human meridians through drugs and heat stimulation to prevent and treat diseases. According to the principle of syndrome differentiation and treatment of traditional Chinese medicine, acupoints are selected on the corresponding parts, which has a definite effect. It is regarded as a characteristic traditional Chinese medicine therapy for bronchial asthma. This protocol elaborates the methods and steps of patient management, material preparation, selection of acupoints, operation, and postoperative nursing to ensure safe and effective moxibustion treatment to significantly improve the clinical symptoms and quality of life of patients with bronchial asthma.

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

Bronchial asthma is a chronic inflammatory disease of the airway which can lead to symptoms such as recurrent wheezing, shortness of breath, chest ...

Standardized Tuina Protocol for Rat DRG Compression Model

Analgesic Effect of Tuina on Rat Models with Compression of the Dorsal Root Ganglion Pain

Neuropathic pain is a prevalent condition that affects 6.9%-10% of the population and results from nerve damage due to various etiologies, such as lumbar disc herniation, spinal canal stenosis, and intervertebral foramen stenosis. Although Tuina, a traditional Chinese manual therapy, has shown analgesic effects in clinical practice for the treatment of neuropathic pain, its underlying neurobiological mechanisms remain unclear. Animal models are essential for elucidating the basic principles of Tuina. In this study, we propose a standardized Tuina protocol for rats with compression of the dorsal root ganglion (DRG), which involves inducing DRG compression by inserting a stainless steel rod into the intervertebral foramen, performing Tuina manipulation with specific parameters of location, intensity, and frequency in a controlled environment, and assessing the behavioral and histopathological outcomes of Tuina treatment. This article also discusses the potential clinical implications and limitations of the study and suggests directions for future research on Tuina.

Author Spotlight: Analgesic Effect of Tuina on Rat Models with Compression of the Dorsal Root Ganglion Pain  

This article presents a manipulation for treating chronic compression of the dorsal root ganglion in rats using Tuina therapy, along with a method for evaluating its effectiveness based on pain behavior and histopathological results.

Neuropathic pain is a prevalent condition that affects 6.9%-10% of the population and results from nerve damage due to various etiologies, such as lumbar ...

Tuina Therapy for Frozen Shoulders in a Rat Model

Tuina in a Frozen Shoulder Rat Model: An Efficient and Reproducible Protocol

Frozen shoulder (FS) is a common condition with no defined optimal therapy. Tuina therapy, a traditional Chinese medicine (TCM) technique used to treat FS patients in Chinese hospitals, has demonstrated excellent results, but its mechanisms are not fully understood. Building on a previous study, this work aimed to develop a Tuina protocol for an FS rat model. We randomly divided 20 SD rats into control (C; n = 5), FS model (M; n = 5), FS model Tuina treatment (MT; n = 5), and FS model oral treatment (MO; n = 5) groups. This study used the cast immobilization method to establish the FS rat model. The effect of Tuina and oral dexamethasone on the glenohumeral range of motion (ROM) was evaluated, and the histological findings were assessed. Our study showed that Tuina and oral dexamethasone were able to improve shoulder active ROM and preserve the structure of the capsule, with Tuina therapy proving to be more effective than oral dexamethasone. In conclusion, the Tuina protocol established in this study was highly effective for FS.

Author Spotlight: Investigating the Effectiveness of Tuina Therapy for Frozen Shoulder 

This study develops an efficient and reproducible Tuina protocol for treating frozen shoulder established in a rat model. This approach will help to study the Tuina therapy treatment method for frozen shoulders.

Frozen shoulder (FS) is a common condition with no defined optimal therapy. Tuina therapy, a traditional Chinese medicine (TCM) technique used to treat FS ...

Establishing and Validating a Chronic Unpredictable Mild Stress Rat Model

Chronic Unpredictable Mild Stress in Rats based on the Mongolian medicine

Depression is a prevalent affective disorder and constitutes a leading cause of global disability. The limitations of current pharmacological interventions contribute to the substantial health burden attributed to this condition. There is a pressing need for a deeper understanding of the underlying mechanisms of depression, making pre-clinical models with translational potential highly valuable. Mongolian medicine, a subset of traditional medicine, posits that disease occurrence is closely tied to the equilibrium of wind, bile, and Phlegm. In this study, we introduce a protocol for the chronic unpredictable mild stress (CUMS) method in rats. Within this framework, rats are subjected to a series of fluctuating, mild stressors to induce a depression-like phenotype, mimicking the pathogenesis of human depression. Behavioral assays employed in this protocol include the sucrose preference test (SPT), indicative of anhedonia-a core symptom of depression; the open field test (OFT), which measures anxiety levels; and the Morris water maze test (MWM), which evaluates spatial memory and learning abilities. The CUMS method demonstrates the capability to induce anhedonia and to cause long-term behavioral deficits. Furthermore, this protocol is more aligned with Mongolian medical theory than other animal models designed to elicit depression-like behavior. The development of this animal model and subsequent research provide a robust foundation for future innovative studies in the realm of Mongolian medicine.

Author Spotlight: Establishing a Rodent Model for Investigating Depression Factors in Traditional Mongolian Medicine

This protocol outlines a chronic unpredictable mild stress (CUMS) model for depression based on Mongolian medical theory, along with methods for validating behavioral tests.