Abstract: Objective We aimed to evaluate the therapeutic effect of Bushen Wenyang Huayu Recipe (BWHR) on a rat model of endometriosis (EM) that involves kidney yang deficiency and blood stasis and determine whether its mechanism of action involves FoxO1-mediated autophagy. Methods Female-specific pathogen-free SD rats aged 8 ~ 10 weeks were randomly allocated to a sham surgery group, a model group, a high-dose BWHR group, a low-dose BWHR group, or a western medicine group (n= 20). A rat model of EM associated with kidney yang deficiency and blood stasis was established by autologous transplantation combined with an ice bath. After confirmation of the successful induction of EM, the high-dose and low-dose groups were administered BWHR, the western medicine group was administered gestrinone suspension, and the sham and model groups were administered the same volume of water by gavage for 3 weeks. Subsequently, the mechanical pain threshold of the rats was measured using a mechanical electronic pain meter, and organ indices were calculated. The histopathological changes in the eutopic endometrium and ectopic foci were evaluated on hematoxylin and eosin-stained sections. The expression of FoxO1, NF-κB, Beclin-1 and LC3 in eutopic endometrium and ectopic lesions was assessed using immunohistochemistry. The mRNA and protein expression of FoxO1, NF-κB, Beclin-1, LC3-I, and LC3-II was measured using Real time PCR and Western Blot, respectively. Results The protein and mRNA expression of FoxO1, NF-κB, Beclin- 1, LC3-I, and LC3-II was significantly higher in the model group than in the control group (P< 0. 05). However, the protein and mRNA expression of FoxO1, NF-κB, Beclin-1 and LC3 in eutopic endometrium and ectopic lesions in the high-dose and low-dose BWHR groups were significantly lower (P< 0. 05). Conclusions FoxO1, NF-κB, and the autophagy related factors Beclin-1 and LC3 may be involved in the development of EMs and have synergistic effects. BWHR may ameliorate EM by reducing the expression of FoxO1 and NF-κB, which inhibits autophagy.

Abstract: Objective To investigate metabolomic changes in the serum of rats with persistent hypertensive SHR and reveal the possible pathogenesis. Methods Six male SHR and six WKY rats were selected at random and defined as the model group and the control group. Rats were given 120 d standard feeding and rat SBP was measured using the BP2000 platform once a week. Rat myocardial tissue was stained with HE and Masson staining and rat serum was detected by UPLC- MS to identify significant metabolites. In addition, a GO and KEGG pathway enrichment analysis was performed. Results The SBP of the model group gradually increased and peaked at aged 20 weeks, which was followed by a plateau period. The SBP of SHR at all time points was much higher than that of the control group (P< 0. 001). HE staining showed that the myocardial tissue of the model group was in a disordered arrangement, and the vessel walls were thicker, while their lumen was narrower. Homogeneous, pink-colored substances were largely found in both the myocardium and around the lumen. Masson staining revealed a very large amount of collagen fiber accumulated in the myocardium of the model group. The serum metabolic patterns and individual differential metabolites of the two groups were evidently separated from each other, and a total of 44 potential landmark metabolites were finally identified, mainly involving neuroactive ligand-receptor interactions, 5-hydroxytryptamine receptors, and arachidonic acid metabolism pathways. Conclusions Myocardial and perivascular fibrosis were identified in SHRs under long-term hypertension. Serum metabolomics of SHR obviously deviated from the normal metabolic pattern, and significant metabolites were primarily involved in regulating vasoconstriction and vasodilation, modulating the activity of the sympathetic nerve, and inhibiting the inflammatory response.

Abstract: Objective We aimed to characterize the relationship between the expression of the Alu gene in a model of human gastric cancer metastasis and the degree of gastric cancer metastasis in tissues and organs and establish a molecular biological method for the early evaluation of gastric cancer metastasis. Methods Alu gene expression plasmids were constructed from the genomic DNA of human gastric cancer cell lines SGC- 7901, MKN45 and the normal gastric mucosal cell line GES1. Alu gene expression in various concentration of gastric cancer cells SGC-7901, which was widely mentioned as human gastric metastasis cell line, was measured by real-time PCR. The relationship between different Alu expression of various concentration was measured by cycle threshold (Ct) value to modelling the standard curve. A human gastric cancer cell xenograft metastasis model was established by the subcutaneous inoculation of SGC-7901 and MKN45 cells into nude mice. The expression of Alu in the resulting tumors in the mice (in the liver, spleen, lung, kidney, and subcutaneous tissues) was then measured by real-time PCR, and a curve for the relationship between the expression of Alu and the degree of tumor metastasis in each tissue was constructed. Subsequently, fresh tumor samples from patients with gastric cancer were subcutaneously inoculated into nude mice to construct a xenograft model, and the relationships between Alu expression in the tissues of the mice and the degree of tumor metastasis in the organs of nude mice was similarly evaluated. Results The number of SGC-7901 cells negatively correlated with the cycle threshold (Ct) value for the Alu gene (R² = 0. 9239). In the xenograft metastasis model, the expression of the Alu gene was higher in subcutaneous tumors than that in lung and liver metastases, and the expression of the Alu gene in lung and liver metastases was higer than normal nude mice. This was consistent with the result of histopathological examination. In the gastric cancer patient xenograft model, if there was no visible metastasis in the organs of nude mice, the expression of Alu gene (Ct 17. 86) was significantly higher(P< 0. 05)than that of normal nude mice (Ct 22. 18), and if there were metastatic lesions that were visible through the naked eye, the expression of Alu (Ct 14. 29) was very significantly higher than that of normal nude mice (P< 0. 01). Conclusions In a model of human gastric cancer metastasis, the expression of the Alu gene positively correlates with the degree of metastasis, namely the higher expression of Alu, the more metastatic tumor cells were present and the more obvious the metastatic foci were in each tissue.

Abstract: Objective By measuring paw withdrawal thresholds (PWTs), paw thickness, paw skin temperature, and paw blood flow in rats, we explored a simple and reliable method for constructing a chronic ischemic limb pain model in rats. Methods Sixteen rats were randomly divided into a sham chronic ischemic limb pain group (sham CILP group, n= 8) and a chronic ischemic limb pain group (CILP group, n= 8). The chronic ischemic pain model group used O-ring lower limbs near the knee joint to slowly pressurize the limb. The compression intensity was 1. 2 ~ 2. 2 N, with compression performed once a day as well as once a week for 30 ~ 40 min for 3 consecutive weeks. The PWTs and degree of swelling, skin temperature, and local blood flow of the two groups of rats before and after modeling were measured at 6, 13 and 20 days after compression. Results After 20 days of modeling, the ipsilateral and contralateral PWTs of the CILP group were significantly lower than those of the sham CILP group (P< 0.01). The ipsilateral paw thickness of the CILP group was significantly thicker than that in the same period in the sham CILP group as well as the contralateral paw thickness of the CILP group ( P< 0.01) over the same period. Furthermore, the ipsilateral skin temperature of the CILP group was significantly lower than that of the sham CILP group (P<0. 01). In addition, the average ipsilateral blood flow of the CILP group was significantly lower than that of the sham CILP group during the same period (P<0.01). Conclusions Long- term low pressure in the lower limbs of rats can effectively cause chronic ischemic limb pain, as demonstrated by the establishment of a rat model of chronic ischemic limb pain.

Abstract: Objective To further study the role of fibroblast growth factor 9 ( FGF9) in neurodevelopment by establishing a mouse model of conditional FGF9 knockout in oligodendrocytes. Methods Olig1-Cre transgenic mice were crossed with FGF9 transgenic mice ( FGF9^{flox / flox} ). Then, female FGF9^{flox / wt} / Olig1-Cre⁺ mice were mated with male FGF9^{flox / flox} mice to obtain oligodendrocyte-specific FGF9 knockout mice (FGF9^{flox / flox} / Olig1-Cre⁺ ) in the F3 generation. To confirm that FGF9 had been conditionally inactivated in the oligodendrocytes alone, genotyping using PCR, western blotting, and laser confocal were used to evaluate the expression of FGF9 mRNA and protein. An initial characterization of the phenotype of the conditional knockout (CKO) mice was then made. Results We confirmed that the expression of the FGF9 mRNA and protein were appropriate in the FGF9^{flox / flox} / Olig1-Cre⁺ mice. Preliminary phenotypic analysis showed that FGF9 CKO mice survived for the same period of time as control mice and were fertile but developed slowly. Conclusions We have successfully established mice with a CKO of FGF9 in oligodendrocytes, which causes growth retardation.

Abstract: Objective To compare the immune response of specific CD8⁺ T cells induced by H1N1 Influenza A virus PR8 strain in the lungs of young and aged C57BL/ 6 mice. Methods Young ( 3 months) and aged ( 24 months) mice were intranasally infected with 490 plaque-forming units of PR8, then 8 days later, they were euthanized and their lungs collected. Fluorescence-activated cell sorting was used to compare the number and functions of viral antigen-specific CD8⁺T cells between the young and aged mice. Surface staining for the influenza virus MHC-I tetramer was performed to count the antigen-specific CD8⁺T cells; and intracellular cytokine staining was used to quantify the cytokines secreted by CD8⁺ T cells in response to influenza-specific peptides, including tumor necrosis factor ( TNF-α ), interferon-γ, interleukin-2, and granzyme B, which are involved in killing by CD8⁺ T cells.Results The proportion of virus-specific CD8⁺T cells in the aged mice was significantly lower than in the young mice. Moreover, the expression of interferon-γ, TNF-α, and interleukin-2 in activated CD8⁺T cells from the aged mice was significantly lower than in the young mice. In addition, the level of granzyme B expression on CD8⁺T cells in the aged mice was significantly lower. Conclusions The number of specific CD8⁺T cells induced by PR8 influenza virus was lower, and their function was impaired in the lungs of aged C57BL/ 6 mice compared with young mice. This implies that the response by specific CD8⁺T cells in the lungs of mice is impaired by aging.

Abstract: Objective We aimed to establish a novel resource consisting of genetically diverse mice and perform a preliminary characterization of the genetic traits of these mice. Methods (1) A multi-parent panel was used to create the Genetic Diversity mice through appropriate crosses. ( 2 ) Whole-exon sequencing was used to analyze the genetic characteristics of the established Genetic Diversity outbred and inbred mice. ( 3) Using the genome sequences of the founders and offspring, we created the chromosome physical maps of these mice and calculated the founder contributions. Results (1) Genetic Diversity inbred mice were characterized by stable inheritance and consistent phenotype and were suitable for rapid phenotypic correlation analysis. (2) Compared with the inbred mice, the Genetic Diversity outbred mice showed superior growth and reproductive performance, and more genetic variation, which led to a “ mosaic-like ” chromosomal pattern. As unique individuals, the outbred mice are ideal for the study of subtle genetic regulatory mechanisms. (3) The founder contributions to the progeny lines were not uniform and genetic preference existed among the offspring. Conclusions Compared with the more conventional inbred mice, the Genetic Diversity mice established by breeding from multiple lines possessed diverse genetic variations. The Genetic Diversity outbred and inbred lines are associated with pros and cons but complement one another, and together represent a novel and powerful tool for the study of complex genetic traits.

Abstract: Objective To determine the importance of gastric inhibitory peptide receptor (GIPR) signaling for insulin resistance and hepatic steatosis in the adipose tissue of mice. Methods Four-week-old adipocyte-specific GIPR knockout (GIPR^{adipo- / -} ) and wild-type (WT) mice were fed a control (CD) or high-fat (HFD) diet for 15 weeks ( n= 8 per group). Changes in body mass were recorded, and the circulating glucose, insulin, and total GIP concentrations were measured after 5, 8, 10, 12 and 15 weeks of feeding. Quantitative PCR was used to measure the changes in the expression of genes involved in inflammation and GIPR signaling in adipose tissue; the volume, mass, and fat content of the liver were quantitatively assessed using ultrasonography; and insulin-stimulated AKT phosphorylation in adipose tissue, liver, and skeletal muscle was evaluated using western blot analysis. Results During HFD-feeding, the body mass of the GIPR^{adipo- / -} mice was significantly lower than that of WT mice, and they also demonstrated lower insulin resistance, liver mass, and hepatic steatosis. In addition, the plasma interleukin 6 (IL-6) concentration was lower in the GIPR^{adipo- / -} mice than in the HFD-fed WT mice. Conclusions GIPR signaling in adipose tissue plays a role in HFD-induced insulin resistance and hepatic steatosis in vivo, and this may be mediated through IL-6.

Abstract:We aimed to characterize the blood reflux, blood compatibility, and biocompatibility of a blood pump after deactivating the pump in an animal in order to facilitate further studies and clinical application. Methods A left- ventricular assist device was implanted into a healthy sheep via a left thoracic approach. After 6 weeks of normal operation, the pump was stopped, as was warfarin administration. The physiological status of the animal was then monitored, and blood biochemical and coagulation testing was performed before the procedure and between 1 and 12 weeks after the pump was deactivated. Ultrasonographic examination was performed 16 weeks after the pump was deactivated. Results The left-sided cardiac assistance model was established successfully, and 6 weeks after deactivation of the pump the sheep displayed normal behavior and normal physiology, with no significant changes in the test result . Ultrasonography showed that the artificial blood vessel was not blocked and there was reverse blood flow 16 weeks after the blood pump was stopped. Conclusions Deactivation of a blood pump was not associated with effects on blood compatibility, physiological state, or organ function in a sheep. The artificial blood vessel was not obstructed, no thrombosis was found, and left-sided cardiac function was not significantly affected. Thus, this blood pump has good compatibility, and the study has provided a reference for the clinical treatment of unexpected pump failure. Further studies should assess the impact of low-dose anticoagulant therapy and repeated deactivation on physiology and cardiac function.

Abstract: Objective To compare the effects of dietary oil content on the physiology and biochemistry of mice to provide a basis for the selection of appropriate oils for inclusion in animal feeds and for human health. Methods Two levels of dietary energy supply from oils were tested in male C57BL/ 6J mice: a 20% ( low-oil energy supply) and 30% (high-oil energy supply), based on the recommendation that dietary energy supply in lipid form should not exceed 30%. Furthermore, each energy supply level was provided as lard (LA), rapeseed oil (RA), olive oil (OL), or a lard and rapeseed blend oil (mass ratio 1 ∶ 1) (BO). After 16 weeks of feeding, the mice’ s growth performance, fat deposition, serum biochemical indices, and blood glucose concentrations were compared. Results Low-OL diet feeding was associated with lower body mass and accumulation of perirenal and epididymal fat (P< 0. 01). High-OL and high-RA feeding were also associated with lower body mass and accumulation of perirenal and epididymal fat (P< 0. 01). The liver index of both the low-OL and high-OL groups was very high (P< 0. 01), and the kidney index was high in the low-RA group (P< 0. 05) and in the high-LA and high-RA groups (P< 0. 01). The circulating total cholesterol content of the high-RA group was significantly lower than that of the other groups (P< 0. 05), and the triglyceride concentration of the high-OL group was the lowest. The high-density / low-density lipoprotein-cholesterol was significantly higher in the low-RA groups than in the other three low-oil energy supply groups (P< 0. 05), but there was no difference between the high-RA and high-OL groups. Glucose and insulin tolerance testing showed that the low-BO and low-OL groups had superior glucose homeostasis to the other low-oil energy supply groups, but there were no significant differences between the four high-oil energy supply groups, and although the final blood glucose concentration in the BO group was slightly higher, there was no difference in the area under the curve. Conclusions A large amount of dietary lard or a lard / rapeseed oil 1 ∶ 1 blend increases body mass and fat deposition in mice. In contrast, both low and high levels of energy provided as olive oil reduce weight gain and fat deposition, but may have deleterious effects on the liver. Feeding of rapeseed oil is preferable for blood lipid profile but may have deleterious effects on the kidney. None of the four kinds of oil had deleterious effects on glucose homeostasis in mice.

Abstract: Objective To characterize the neuroprotective effect of tanshinone IIA (Tan IIA) and its effects on the phosphatidylinositol-3-kinase / protein kinase B (PI3K/ AKT) pathway in a mouse model of Alzheimer’s disease (AD). Methods A mouse model of AD was prepared by the lateral intracerebroventricular injection of lipopolysaccharide (LPS). Mice with AD were randomly allocated to a model group, and low-dose, medium-dose, and high-dose Tan IIA groups, and untreated mice were used as a control group. Tan IIA treatment groups were administered Tan IIA intraperitoneally at 1, 5, or 10 mg / kg, once daily for 7 d. The learning and memory of the mice in each group were assessed using the Morris water maze, Y-maze, and autonomous activity tests. Pathological changes in the brain were assessed using hematoxylin and eosin- stained sections. The concentrations of brain-derived neurotrophic factor (BDNF) and cholinergic and inflammatory factors in brain tissue homogenates were measured by ELISA. The expression of ionized calcium binding adaptor molecule-1 (IBA- 1) and glial fibrillary acidic protein (GFAP) in brain tissues were assessed using immunohistochemistry. The expression of nuclear factor κB (NF-κB), toll-like receptor 4 ( TLR4), PI3K, and AKT was measured by Western Blot. Results There were higher AchE, IL-6, and TNF-α, IBA-1, GFAP, p-NF-κB/ NF-κB and TLR4 concentrations in the brain tissue and escape latency in the model group than in the control group (P<0. 05); whereas the free alternation, and BDNF, Ach, p-PI3K/ PI3K, and p-AKT/ AKT concentrations were significantly lower ( P< 0. 05 ). The escape latency and the concentrations of AchE, IL-6, TNF-α, IBA-1, GFAP, p-NF-κB/ NF-κB, and TLR4 in brain tissue were significantly lower in the Tan IIA treatment groups than in the control group (P<0. 05), whereas the free alternation and BDNF, Ach, p- PI3K/ PI3K, and p-AKT/ AKT concentrations in brain tissue were significantly higher (P< 0. 05). The effects of the high dose of Tan IIA were the most marked. However, there were no significant differences in the autonomous activity distances among the groups (P> 0. 05). Conclusions Tan IIA protects mice with AD from nerve damage, improves their learning and memory, which may be achieved via activation of the PI3K/ AKT pathway and a reduction in brain inflammation, which would inhibit the excessive activation of microglia and astrocytes.

Abstract:To evaluate the toxicity of circulating acrylamide in mice. Methods Mice were gavaged daily with acrylamide solution for 16 days. Their body and organ masses were recorded and relevant blood parameters were measured. Results Doses of 120 or 150 mg / kg acrylamide daily caused significant reductions in body mass gain, relative liver mass, WBC, LYM, RBC, MCHC, PLT, and P-LCR of the mice ( P< 0.05), whereas GRAN increased significantly ( P< 0.05). Doses of acrylamide of 90, 120, and 150 mg / kg caused significant reductions in the relative kidney mass and HGB (P< 0.05). A dose of 150 mg / kg significantly reduced MCV and PCT (P< 0.05). Doses ≥ 60 mg / kg caused significant increases in AST, ALT, AKP, and CREA (P< 0.05), and when the concentration was 60 mg / kg, the AST activity was highest. Doses ≥ 90 mg / kg caused significant increases in UA and BUN (P< 0. 01), and when the dose was 120 mg / kg, the BUN was highest. Conclusions Acrylamide has toxic effects in mice. Medium and high doses of acrylamide (≥ 90 mg / kg) have deleterious effects on red blood cells, white blood cells, the liver, kidney, and other organs in mice. High concentrations of acrylamide (≥ 120 mg / kg) also have significant deleterious effects on platelets.

Abstract: Objective Establish an orthotopic and subcutaneous patient-derived xenografts ( PDX) model of hepatocellular carcinoma (HCC) and compare their growth and pathological characteristics. Methods Fresh tumor tissues from HCC patients from Beijing Ditan Hospital were collected, and implanted in NCG immunodeficiency mice liver and subcutaneous tissues. Their cell growth, pathological characteristics, tumor formation rate, and tumor formation time were analyzed carefully. Results This study established orthotopic and subcutaneous PDX model successfully, and both of them were consistent with the primary tumor pathological characteristics. The tumor formation rate were 50% ( 3 / 6) and 20% (3 / 15) in orthotopic and subcutaneous tissues, respectively. The average tumor formation time were 30 and 68 days in the orthotopic PDX model. Conclusions The orthotopic model has a higher tumor formation rate, it provides a new choice for difficult tumor growth. The subcutaneous model were easy to operate and observe. Hence, the two models were choose according to the specific scientific research practice. They provides effective animal models for further exploration of the pathogenesis of HCC and drug screening.

Abstract:Systemic lupus erythematosus encephalopathy (NPSLE) is a nervous system complication of systemic lupus erythematosus (SLE). Various animal models of NPSLE have been developed that are characterized by histological defects, such as neuronal apoptosis and fewer synapses; brain leukocyte infiltration; and behavioral defects, such as emotional abnormalities and impaired learning and memory. In this article, we summarize the method used to construct animal models of NPSLE, describe the principal characteristics of each, and discuss the use of these models in the study of the pathogenesis of the disease and the identification of potential therapeutic targets. These models are useful means of defining the pathogenesis of NPSLE, screening potential therapies, and developing techniques for personalized diagnostic and treatment plans.

Abstract:Irritable bowel syndrome ( IBS) is a common intestinal disease characterized by recurrent abdominal pain and changes in the feces and / or defecatory habits. However, although the etiology and pathogenesis of IBS remain unclear, visceral hypersensitivity is widely accepted to be an important pathogenetic factor. Therefore, it is important to be able to study an appropriate animal model of visceral hypersensitivity. Colorectal distension (CRD) is widely used for this purpose because it induces persistent and stable visceral hypersensitivity through continuous mechanical stimulation that is not associated with obvious pathological changes in the rectum, which is consistent with the pathological characteristics of IBS. In the present paper, we discuss the surgical procedure, the limitations of CRD, the various experimental devices and expansion method used, and other related factors in order to provide a reference for researchers who wish to use CRD-based models of visceral hypersensitivity.

Abstract:Cardiorenal syndrome (CRS) is the end-stage form of heart and kidney disease and it is associated with a poor outcome. Further study of the physiological and pathological mechanisms of CRS is still required to facilitate improvements in treatment. Therefore, it is important to establish appropriate animal models that simulate the features of the pathogenesis of the disease in humans. In this article, we have reviewed and evaluated the clinical relevance of animal models of type II CRS on the basis of a search of the international literature. We discuss simple animal models, which include coronary artery ligation, aortic constriction, spontaneous hypertension, drug administration, arteriovenous fistula, and transgenic mouse models; and composite models, including coronary artery ligation combined with nephrectomy, and nephrectomy combined with drug administration.

Abstract:Vascular cognitive impairment (VCI) has been a hot topic in medical research in recent years. The cause of the disease is relatively complex, but the main clinical manifestations are multiple cognitive disorders, impaired short-term memory, and dementia, which seriously affect quality of life. The successful establishment of animal models is an important step in the study of the pathogenesis of VCI and as a means of identifying effective treatment options for patients. Numerous animal models of VCI have been established, of which rodent models are the most widely used. In rats, these include a four-vessel occlusion (4-VO) method, a modified 4-VO method, a three-phase 4-VO method, a two-vessel occlusion ( 2-VO) method, a modified 2-VO method, a modified common carotid stenosis method, and a one-sided common carotid arterial occlusion method. In mice, these include a common carotid artery stenosis method and an asymmetric common carotid artery surgery method. These surgical modeling method are associated with differing postoperative site of injury, severity of injury, survival rate, and behavior. We will systematically review the surgical method, phenotypes, evaluation, advantages, and disadvantages of the nine listed rodent VCI models. We hope that this will provide some guidance for researchers in the selection of relevant animal models.

Abstract:Huntington’ s disease ( HD) is an inherited autosomal dominant and progressive neurodegenerative disease with locomotor, cognitive, and psychiatric functional deficits. Currently, there is no cure for HD, and the clinical benefit is greatly limited for some treatments available for controlling HD symptoms. In 1993, it was discovered that HD is caused by a single Huntingtin gene (HTT) mutation with 36 CAG repeats and or more, which translates to a mutated polyglutamine, Resulting in neuronal excitotoxicity and abnormal mitochondrial energy metabolism, and eventually neurodegeneration in the cerebral cortex and basal nuclei. To better understand the pathogenesis mechanisms of HD and explore therapeutic interventions, it is important to create HD animal models. In this review, we will introduce HD animal models based on neuropathology, and explore the molecular biology of HD such as neural excitotoxicity and mitochondrial toxicity.