Abstract: Objective To investigate the effect of low-molecular-weight heparin on the EGFR/ PI3K/ Akt signaling pathway, to explore its effect on endometrial angiogenesis during blastocyst implantation, and to evaluate the mechanism of the drug’s effect on endometrial receptivity. Methods Seventy-two C57BL/ 6 mice were randomly divided into six groups: blank group, model group, aspirin group, low-molecular-weight heparin high-dose group, low-molecular-weight heparin medium-dose group, and low-molecular-weight heparin low-dose group. Apart from the blank group, in the remaining groups indomethacin was used to establish a blastocyst implantation disorder model. On the morning of the fifth day of pregnancy, the mice were sacrificed by cervical dislocation and the number of uterus-implanted blastocysts was counted. The expression of HBEGF, EGFR, PI3K, and Akt in endometrium was determined by immunohistochemistry, Western Blotting, and Real-time PCR. Results Compared with the model group, the mice in the blank group and the drug treatment group had uteruses with ruddy, uniformly distributed implants, and a greater number of implanted sites ( P< 0. 05). The endometrial HE-stained epithelial cells were thick, the interstitial cells were large and compactly arranged, and there were more glands and blood vessels. The protein and mRNA expression levels of HBEGF, EGFR, PI3K, and Akt in the endometrium with low molecular weight heparin and high dose was higher than that of each group ( P < 0. 05). Conclusions Low-molecular-weight heparin can promote the expression of HBEGF and activate the EGFR/ PI3K/ Akt signaling pathway, thereby improving endometrial receptivity and increasing the success rate of blastocyst implantation.

Abstract: Objective To establish a heart failure model in mice, and to use ultrasound imaging and pathology techniques to comprehensively evaluate the characteristics of cardiac function and structural changes during disease development. Methods The transverse aortic constriction ( TAC) surgical technique was used to establish a model of pressure-overload-induced heart failure in mice. Blood flow Doppler spectra were collected to evaluate the pressure at the aortic arch stenosis before surgery and at weeks 2, 4, and 8 after it. B-mode and M-mode ultrasound images were collected to evaluate cardiac structure and systolic function. The blood flow Doppler spectrum from the mitral valve orifice and tissue Doppler spectrum from the mitral annulus were combined to evaluate cardiac diastolic function, and cardiac tissues were collected for histopathological observation. Results The ultrasound result showed that the blood pressure of the aortic arch stenosis increased significantly at weeks 2, 4, and 8 after the operation. At 2 weeks after the operation, the thickness of the left ventricular wall typically showed compensatory increases, while the inner diameter was decreased, accompanied by impaired diastolic function and myocardial fibrosis. Four weeks after the operation was the transition period, which was mainly manifested as a transition of the left ventricle from hypertrophy to dilation, decreased systolic function, continuously impaired diastolic function, and further fibrosis in the myocardium. Eight weeks after surgery, the ventricle was characterized by cavity enlargement, significant reductions of systolic and diastolic function, and cardiomyocytes being partially lysed and significantly fibrotic. Conclusions By evaluating the structure and function of the left ventricle in the three stages of compensated hypertrophy, transitional stage, and decompensated dilatation of a TAC-induced heart failure model in mice, the use of the model should advance basic research and translational medicine.

Abstract: Objective To observe the therapeutic effects of electroacupuncture (EA) on mechanical pain of a CRPS-I rat model. To study the intervention of EA on the oxidative stress response from local tissues of a CRPS-I rat model and further investigate the mechanisms underlying acupuncture analgesia. Methods The chronic post-ischemic pain model (CPIP) was established to mimic CRPS-I. Healthy male SD rats were randomly divided into Sham, CPIP, CPIP + EA, and CPIP + Sham EA groups. After model establishment, EA was applied on a daily basis. The 50% paw withdrawal thresholds (PWTs) were measured by the von Frey hair test. Detection kits were used to measure the oxidative stress- related markers. Immunostaining was used to detect cellular oxidative damage. Results 1) Compared with those in the Sham group, the 50% PWTs were significantly decreased in CPIP group rats (P< 0. 01) and lasted until the end of the experiment, indicating the successful establishment of the CPIP rat model. 2) After EA treatment, the 50% PWTs were significantly increased in the CPIP + EA group ( P< 0. 01), compared with those in the CPIP + Sham EA group. 3) Compared with the Sham group, the CPIP group showed significantly decreased levels of SOD and GSH and increased levels of H2O2 , MDA, and 8-OHG in ipsilateral hind paw tissues (P< 0. 05 or P< 0. 01). 4) After EA treatment, compared with the CPIP + Sham EA group, the CPIP + EA group showed significantly increased levels of SOD and GSH and decreased levels of H2O2 , MDA, and 8-OHG (P< 0. 05 or P< 0. 01). Conclusions Our result indicate that EA can significantly reduce the oxidative stress response in ipsilateral hind paw tissues of CPIP model rats. This mechanism may be involved in EA’s therapeutic effect on mechanical pain of CPIP model rats.

Abstract: Objective The purpose of this study was to explore a method of coronary heart disease model of qi- deficiency and blood-stasis syndrome type in rats by comparing the tongue appearance and its relationship with prostacyclin (PGI₂) and thromboxane A2 (TXA₂) of three different model estalishment method ,which were respectively established by pathological factors, etiological factors and compoundfactors. Methods Thirty-two rats were randomly divided into four groups: Group K:Normal food and water ( n = 8); Group L: normal feeding twenty-one days and then ligation the left anterior descending coronary artery (n = 8); Group Y:control diet combine with exhautive swimming for twenty-one days (n = 8); Group F: control diet combine with exhautive swimming for twenty-one daysand then ligation the left anterior descending coronary artery ( n = 8). At the end of modeling, Photoshop 6. 0 was used to analyze the RGB value of the tongue surface and an RGB data distribution range table was used to evaluate the tongue color of each model. HE staining was used to determine the numbers of microvessels in the upper cortex, keratinized layer, lamina propria, and lamina propria. Immunohistochemistry was used to analyze the positive expression rate of PGI₂ and TXA₂, and calculate the T/ P ratio. Results (1) The tongue quality in Group K was in the category of “light red tongue”. In Group L, the R value of the tongue surface was significantly lower than that in Group K, reflecting “dark red tongue”. In Group Y, the R value of the tongue surface was also significantly lower than that in Group K, reflecting “ light white tongue”. In Group F, the R value, G value, and B value of the tongue surface were significantly lower than those in Groups K and L, reflecting “purple tongue”. (2) Compared with Group K, there were no differences in the height of each layer and the number of microvessels of the filiform papilla on the lingual surface in Group L. Compared with Group K, there was a significant decrease in the height of the keratosis layer of filiform papilla in Group Y. In Group F, there were significant differences in the height and number of microvessels between Group K and Group L, while there was no difference in the height of the lamina propria in each group. (3) Compared with that in Group K, there were no significant changes of PGI₂ in Groups L, Y, and F. The TXA₂ level and the ratio of T/ P in Group L were significantly higher than those in Group K. The TXA₂ level and the ratio of T/ P in Group F were significantly higher than those in Group K and Group L. Conclusions The RGB quantitative characteristics of tongue color of CHD with qi deficiency and blood stasis syndrome are related to the decrease of the height of the suprapapillary cortex and keratodermis, and the increase of the number of capillaries in the lamina propria. The molecular biological mechanism may be related to the imbalance of TXA₂ / PGI₂ . The pathological etiology complex model is more consistent with the characteristics of CHD with qi deficiency and blood stasis syndrome.

Abstract: Objective To establish a model of lung metastasis by bone marrow injection and a model of tumor recurrence after surgery to reveal the superiority and feasibility of the model of bone marrow injection to provide a new research basis for studying lung metastasis of malignant tumors. Methods The mice were divided into a mammary fat pad injection group, a subcutaneous injection group, a tail vein injection group, and a bone marrow injection group. Different models of 5 × 105 4T1 cells were injected via mammary fat pad injection, subcutaneous injection, tail vein injection, and bone marrow injection, and the growth, survival period, and lung metastasis efficiency of the primary tumors of model mice under different modeling method were observed. Different numbers of 4T1 cells were injected and the time of lung metastasis was observed. 4T1-luc cells were injected into the bone marrow to construct a postoperative tumor recurrence model. The mice were divided into a sham amputation group, a 3rd day amputation group, a 7th day amputation group, and a 10th day amputation group. Live imaging was performed every 7 days to observe lung metastasis in mice, while imaging was performed in the lungs of mice in the pseudo-amputation group and the 7th day amputation group to confirm the success of model construction. Results Compared with mammary fat pad injection and subcutaneous injection, bone marrow injection had no effect on the growth of the primary tumor, but the associated survival time was significantly shortened. Compared with mammary fat pad injection, subcutaneous injection, and tail vein injection, bone marrow injection had the highest lung metastasis efficiency. The bone marrow injection method required only 1 × 105 4T1 cells to cause lung metastasis on the 12th day. Mice with bone marrow injection still exhibited lung metastasis and recurrence of primary tumor after amputation. Conclusions A mouse model of lung metastasis can be successfully established by bone marrow injection along with a mouse model of tumor recurrence after surgery. The mouse model created using this injection method has the characteristics of a short survival time and high metastasis efficiency. It can be used to study the mechanism of lung metastasis of malignant tumors and the related screening of drugs for treating malignant tumors.

Abstract: Objective To explore the protective potential and mechanism of angiotensin-converting enzyme II (ACE2) in acute lung injury (ALI) induced by limb ischemia reperfusion in mice. Methods Male wild-type and ACE2 transgenic mice (overexpression of ACE2) Institute of Cancer Research mice were randomly divided into six groups (n =18): wild control group (Control), wild model group (Model), ACE2 control group (ACE2 + Control), ACE2 model group (ACE2 + Model), ACE2 model + A779 group (ACE2 + Model + A779), and ACE2 model + MLN-4760 group (ACE2 + Model + MLN-4760). ALI models were established using rubber band ligation of the bilateral hind limb roots (ischemia for 2 h and reperfusion for 4 h). Lung histological changes were observed using hematoxylin and eosin (HE) staining. Lung water content and pulmonary permeability were indicated by the organ coefficient, wet-to-dry weight ratio, cell numbers, and protein concentration in the bronchoalveolar lavage fluid (BALF). Enzyme-linked immunosorbent assay was used to determine the concentrations of BALF interleukin ( IL) - 6, tumor necrosis factor ( TNF) - α, and lung angiotensin II (Ang II) / angiotensin-(1-7) [Ang-(1-7)]. Quantitative real-time polymerase chain reaction was used to analyze mRNA expression of ACE/ ACE2, and western blot used to quantify the protein expression of ACE/ ACE2 and AT1 / Mas receptors. Results Compared with the wild model group, overexpression of ACE2 attenuated lung lesions ( HE staining and lung injury score), reduced alveolar capillary permeability (organ coefficient, wet-to-dry weight ratio, BALF cell numbers, protein concentration), improved the expression profiles of inflammatory cytokines (IL-6 and TNF-α) in the BALF, and (particularly) reversed pulmonary renin-angiotensin system imbalance. Moreover, these effects were abrogated by MLN-4760 (a specific ACE2 inhibitor) and A779 ( a specific Mas receptor antagonist). Conclusions The findings indicate that ACE2 can ameliorate the imbalance of the pulmonary renin-angiotensin system and the ALI via the ACE2-Ang- (1-7)-Mas axis.

Abstract: Objective To prepare Clostridium perfringens (CVCC60101) for production and testing of veterinary vaccines, verify its biological characteristics, and perfect the standard and method of preparation and verification. Methods In this study, Clostridium perfringens (C) prepared and stored by freeze-drying in 1960 was resuscitated, rejuvenated using susceptible animals, isolated, cultured, freeze-dried, and the new strain’s morphology, culture characteristics, biochemical properties, serological characteristics, virulence, immunogenicity, and other characteristics were tested. Results The result showed that the new strain was pure, and its morphology, culture characteristics, and biochemical properties accorded with the characteristics of Clostridium perfringens. In addition, its vacuum degree, residual moisture, serological characteristics, virulence, and immunogenicity accorded with the standard of the strain for production and testing of veterinary vaccines. Conclusions This research gives more experimental data for perfecting the standard and method of preparation and verification of Clostridium perfringens (C).

Abstract: Objective To determine the histological difference between lesions induced by balloon injury and spontaneously formed plaque and to explore the potential impact on scientific research related to these differences. Methods Using the CNKI, Wanfang, Weipu, PubMed, and Web of Science databases, relevant studies on the coronary artery lesions induced by balloon injury were selected. Pathological images of atherosclerotic plaques in the articles were selected and analyzed for their differences from spontaneously forming atherosclerosis. Results Plaques induced by the balloon injury method can often form severe intimal hyperplasia in a short period of time, Resulting in stenosis of the lumen. The main components of the hyperplastic intima are smooth muscle cells and proteoglycans. Internal elastic plate rupture is common, while fibrosis cap and necrotic cores are rare. Conclusions There are differences in morphology between intimal hyperplasia caused by balloon injury and atherosclerosis. Balloon injury may not be suitable as an atherosclerosis model and for mechanistic research and preclinical evaluation.

Abstract: Objective We aimed to improve an existing modeling method to establish an experimental model suitable for cancer research with short experimental period, low mortality, and modeling cost for metastasis. Methods Animals were injected intraperitoneally with 200 mg / kg N-diethylnitrosamine (DEN) at the beginning of the experiment. All animals in the first and second groups were given 40 or 80 ppm N-nitrosomorpholine (NMOR) via drinking water for 20 or 10 weeks. Animals in a third group were given 0. 8 mg / kg NMOR via gavage for 17 weeks. Animals in the second and third groups were fed for 7 weeks following the NMOR treatment. All animals in the first and third groups were subjected to two-thirds partial hepatectomy ( PH) to enhance cell proliferation activity at the end of the second week after DEN treatment. Results The first group had a liver tumor incidence of 10%, a survival rate of 100%, and a lung metastasis rate of 0% at 20 weeks. The second group had a liver tumor incidence of 35%, a survival rate of 76%, and lung metastasis rate of 0% at 17 weeks. The liver tumor incidence was 100% in the third group. The survival rate was 100% or 55% at 20 or 24 weeks. The lung metastasis rate was 55%. Conclusions Overall, these result suggest that modeling method such as intragastric administration of 0. 8 mg / kg NMOR for 17 weeks after intraperitoneal injection of 200 mg / kg DEN and a two- thirds PH can effectively establish an advanced liver tumor model with a high survival rate and morbidity in 20 weeks. In addition, metastases can appear at the later stages. This modeling method has several advantages, such as high incidence of hepatocellular carcinoma, high model survival rate, and late-stage metastasis. This could be an ideal model for research on the prevention and mechanism of hepatocellular carcinoma.

Abstract: Objective To explore the role of the p21 gene in the migration and invasion of normal trophoblasts and the possible associated mechanisms. Methods The p21 gene in trophoblast HTR- 8 / SVneo was knocked down by siRNA. Cell proliferation, wound assays, and Transwell invasion assays were used to observe changes in the migration and invasion of trophoblasts with p21 knockdown. The effects of p21 knockdown on ERK3 and MMP2 expression were also examined at both mRNA and protein levels by real-time fluorescent quantitative PCR (qPCR) and western blotting. Results The migration and invasion of trophoblasts with p21 knockdown were significantly reduced, whereas there was no effect on cell proliferation. The expression of ERK3 and MMP2 was significantly decreased at both mRNA and protein levels. Conclusions These result indicate that the p21 gene can promote the motility of normal trophoblasts, and this effect is related to regulation of the expression of ERK3 and MMP2.

Abstract: Objective To observe and compare the effects of gender and injection doses on isoproterenol ( ISO) induced cardiac hypertrophy model in rats. Methods Male and female SD rats were randomly divided into three groups as follows: normal control group, low dose of group (1. 25 mg / kg) and high dose of group (5 mg / kg). Ten days after the subcutaneous injection of ISO, the differences in the parameters from echocardiography, hemodynamics, histopathology and Real-time PCR were compared. Results (1) After treatment with two doses of ISO, various parameters of heart weight, interventricular septum thickness, myocardial cell area, myocardial fibrosis and mRNA expression levels of ANP and BNP were significantly increased (P< 0. 01), while left ventricular function was significantly reduced in rats (P< 0. 01). (2) The changes of left ventricular weight / body weight, interventricular septum thickness, left ventricular end-systolicpressure (LVESP), left ventricular pressure maximal rate of fall (-dp / dtmax ) and ANP mRNA expression were significantly increased or decreased in a dose-dependent manner in male rats ( P < 0. 01). However, only the left ventricular end- diastolicpressure(LVEDP),left ventricular pressure maximal rate of fall( dp / dtmax) was decreased in a dose-dependent manner in female rats. (3) Female rats exhibit a severe reduction in cardiac function than male rats when injected with low- dose of ISO (P< 0. 01). In contrast, male rats exhibited a significant increase in left ventricular weight / body weight and ANP mRNA expression than female rats when injected with high-dose of ISO (P< 0. 01). (4) In male rats, the survival rate of low- and high- dose of groups of ISO were 63. 6% and 81. 8%, respectively. The survival rate of low- and high- dose of groups in female rats were 63. 6% and 55. 0%, respectively. Moreover, no rats died in the normal group. Conclusions The dose of injection has a greater effect on the degree of cardiac hypertrophy in male rats induced by ISO. It is best to choose a dose of 5 mg / kg for male rats and 1. 25 mg / kg for female rats to establish an ISO-induced cardiac hypertrophy model.

Abstract: Objective To establish a rat model that can exhibit pathological changes of the sacral ligament similar to that in patients with pelvic organ prolapse, and to explore the effects of increasing the intensity of labor injury and simulating menopause on the histopathology of the sacral ligament in rats, so as to provide an experimental basis for further research. Methods Sixty adult SPF female rats with a body weight of about 300 g were selected, 45 of which had no birth history. They were divided into a blank control (a) group, simulated menopause (b) group, and simulated birth injury + simulated menopause (c) group by a random digital table method. The other 15 rats who had delivered three times in succession were used as alabor injury + simulated birth injury + simulated menopause (d) group. Group A received normal drinking water and food, group B received bilateral ovariectomy, group C received simulated labor injury operation and bilateral ovariectomy to simulate menopause, and group D received bilateral ovariectomy to simulate menopause on the basis of simulated labor injury twice after three consecutive deliveries, which were fed for 8-10 weeks. Eight rats were randomly selected from each group to observe the appearance of the genitalia and the changes of the genital cleft. The changes of integral optical density of type I and type III collagen (COL1A1, COL3A1) and transforming growth factor β-3 (TGFβ-3) in the anterior wall of the vagina were observed and evaluated by immunohistochemistry. The expression levels of COL1A1, COL3A1, and TGFβ-3 mRNA in the sacral ligament were measured by RT-PCR. Results After modeling, the diameter of the vaginal hiatus in group D was more than 2 mm greater than that in the control group, and there was no obvious prolapse in each group, while group D showed a slightly abnormal perineum.The expression of COL1A1 in the anterior wall of the vagina in group D was significantly lower than that in group A (P< 0. 05), and the relative expression of TGFβ-3 in group C and group D was significantly higher than that in group A (P< 0. 05). The expression of COLIA1 and COL3A1 in the sacral ligament in group D was significantly higher than that in group A (P< 0. 05). The relative expression of TGFβ-3 in group C and group D was significantly higher than that in group A (P< 0. 05). Conclusions After the establishment of the model, the phenotypic outcomes of pelvic organ prolapse was not notable. This model is not suitable for the study of phenotypic changes. This model can be used as an animal model to study the pathophysiological changes of the sacral ligament of pelvic organ prolapse, but it cannot be determined whether the mechanical properties of the sacral ligament of human and rat are similar. It is necessary to combine the advantages and disadvantages of various animals and the actual issue to be resolved to select the most suitable animal model for research.

Abstract: Objective The aim of the study was to evaluate the relationship between portal hypertension and ligation of left portal vein ( LLPV), together with the possible role of NO in prehepatic portal hypertension ( PHT). Methods Thirty-six 8-week-old SD rats were divided into six groups: LLPV with needles of 0. 6, 0. 7, or 0. 8 mm in diameter; ligation of the main portal vein (LMPV) with a needle of 0. 9 mm in diameter; left lateral lobe of liver ligated and removed (LLLR); and sham operation. The portal vein pressure (PVP) was measured in all rats before the operation, immediately after the operation, and 15 and 30 days after the operation. All rats were sacrificed 30 days after the operation. The portal vein blood, and left and right lobes of liver tissues were saved to measure the concentration of NO. Results For rats in the LLPV group, the PVP levels immediately, 15 days, and 30 days after the operation were all significantly higher than the preoperative PVP and PVP of the sham operation group at the same time. No statistically significant difference of PVP was found between the LLPV group with a needle of 0. 6 mm in diameter and the LMPV group 30 days after the operation. There were also no significant differences of the NO concentration in left and right lobes of liver tissue and blood in all groups. Conclusions LLPV can also form stable PHT. A needle of 0. 6 mm in diameter can acquire good result . NO does not play a role in prehepatic PHT.

Abstract:Animal experiments are essential for medical research, which has led to the sacrifice of many laboratory animals (LA). However, various emerging techniques can be used to replace experiments requiring LA sacrifice or products of LA sacrifice. In many instances, these techniques provide more reproducibility and parameter control than experiments involving LA or LA products, and in particular can reduce the sacrifice of LA and avoid issues with ethical approval during the experiment. In silico simulation, informatics, 3D cell culture models, new types of cell culture medium, and products of animal-friendly affinity reagents are some innovative technologies that can reduce the sacrifice of LA, reduce the cost of experiments, and ensure the accuracy of experimental result . At the same time, owing to species differences between animals and humans, the reliability of some experimental result related to human health has also been questioned. Finally, alternatives to animal experiments help to reduce the number of animals required for research to a certain extent, but cannot completely eliminate the need for animals in research. This article summarizes the progress of related alternative technologies in recent years, provides ideas for the development of relevant experiments in medical research, achieves the purpose of reducing LA sacrifice, and improves the welfare of LA.

Abstract:Crohn’s disease is a chronic inflammatory bowel disease, the pathogenesis of which has long been a hotspot of medical research but remained difficult to clarify. The selection of suitable animal models for the study of Crohn’s disease is particularly important to discover potential therapeutic targets and develop new therapeutic options. As biotechnology advances, the animal models available for studying Crohn’ s disease are becoming more diverse, from the original chemically induced animal models to later spontaneous animal models, cell transplantation animal models, to the widely used genetically engineered animal models available today, giving researchers more and more suitable options. This paper provides an overview of the pathogenesis and advantages and disadvantages of these animal models, providing a reference for basic research.

Abstract:Pluripotent stem cells (PSCs) are a class of stem cell lines that can proliferate indefinitely and have the potential to differentiate into various tissue cells. The establishment of induced pluripotent stem cells (iPS cells) has further narrowed the distance between stem cells and clinical disease treatment, and has great potential value for applications in the fields of agricultural economic development and animal medicine. However, currently the thorough mechanism research and the application of stem cells are mainly focused on mouse and human stem cells, while the research on stem cells of large animals with a close relationship with agriculture, animal husbandry, and veterinary science is still at the preliminary establishment and application trial stage. By summarizing the relevant literature, this article briefly describes the establishment and research status of embryonic pluripotent stem cells and induced pluripotent stem cells in large animals such as pigs, cattle, and horses.

Abstract:As a physiological barrier for restricting material exchange between blood and brain, blood brain barrier prevents harmful substances from entering the brain and maintains the stability of the central nervous system. In recent years, researches on the structure and function of the blood brain barrier among different species has progressed largely, which help to reveal the evolution of the blood brain barrier and to understand the specific mechanism regulating. This article will review the structure and function of the blood brain barrier of multiple animals including fish, birds, and mammals in recent studies.

Abstract:Impaired immunity makes the body more susceptible to internal and external pathogenic factors, making people more susceptible and at higher risk for various diseases. To better conduct basic research for this special group, based on relevant studies performed in China and elsewhere in the past 10 years, this paper analyzes and summarizes the experimental animals, Methods and evaluation indexes involved in acquired secondary immunodeficiency models, and proposes that immunocompromised models can be integrated into future research on the modernization of traditional Chinese medicine.

Abstract:Diabetes mellitus (DM) has become a worldwide epidemic, with an increasing incidence year by year. To further explore the pathogenesis of DM and develop more effective prevention and treatment programs, researchers need stable, replicable animal models that are similar to human diseases. The administration of streptozotocin ( STZ) has been used for half a century to induce DM, and is the most widely used method for establishing animal models of DM in basic research. In this paper, the main factors influencing STZ-induced DM rat models are briefly reviewed. On the basis of the research result and related literature, some suggestions for the future preparation of STZ-induced DM rat models are made.

Abstract:Primary headache is a condition with a high disability rate, particular in young adults. It not only brings suffering to the patients but also places a heavy burden on society. However, the pathogenesis of primary headache is still unclear. The establishment of reliable animal models of primary headache is helpful to explore the pathogenesis and develop drugs for prevention and treatment. Here, we comprehensively summarize animal models of primary headache in recent years from the perspectives of animal model principles, Methods evaluations, scope of application, and advantages and disadvantages.