Abstract: Objective To investigate the protective effect of hederagenin (HDG) on cisplatin (Cis)-induced acute kidney injury (AKI) in mice and its potential mechanism. Methods 24 male C57BL / 6J mice were randomly divided into a control group, AKI model group, HDG low-dose group, and HDG high-dose group, with six mice in each group. AKI model was established by intraperitoneal injection of 20 mg / kg cisplatin(Cis). The HDG low-dose and HDG high-dose groups were given 20, 40 mg / kg HDG by intragastric administration, respectively, and samples were collected 3 days later. The kidneys of the mice were collected for hematoxylin-eosin (HE) and periodic-acidschiff (PAS) staining to evaluate the kidney pathology, and serum was collected to detect changes in serum creatinine(Scr) and blood urea nitrogen ( BUN). The expression of p-P65, P65, IL-6, TNF-α, IL-1β, and other inflammatory-related proteins was detected by Western Blot. A TCMK1 ( renal tubular epithelial cell) inflammatorycell model was established by Cis (200 ng / mL) stimulation in vitro. Blank group, Cis model group, HDG low-dose group, HDG high-dose group, Axin2 overexpression group, HDG + Axin2 overexpression group were set up. In the Axin2-overexpression group, the expression of p-P65, P65, IL-6, TNF-α, IL-1β, Axin2, and AREG was detected among total cell proteins. Results Compared with the control group, AKI model mice exhibited significantly elevated serum creatinine and blood urea nitrogen levels ( P<0. 05), accompanied by pathological alterations including vacuolar degeneration of renal tubules, inflammatory cell infiltration, and glycogen deposition, and the expression of inflammation-related proteins (p-P65, TNF-α, IL-6, IL-1β) and Axin2 was markedly upregulated in AKI mice (P<0. 05). HDG treatment induced a dose-dependent reduction in serum creatinine and blood urea nitrogen levels (highdose > low-dose, P<0. 05), alleviated renal histopathological damage, and concurrently suppressed the expressionof these inflammatory mediators and Axin2 (P<0. 05). HDG was confirmed that dose-dependently inhibited Cisinduced upregulation of Axin2, and inflammatory cytokines in vitro experiments. Transcriptome sequencing revealed that Axin2 overexpression significantly increased amphiregulin (AREG) expression (P<0. 05). Mechanistically,HDG reduced p-P65 phosphorylation by suppressing the Axin2 / AREG axis (P<0. 05), while Axin2 overexpression abolished the protective effects of HDG against Cis-induced renal tubular cell injury. Conclusions HDG protects against renal injury in AKI mice by reducing inflammation through the inhibition of Axin2 / AREG axis activation.

Abstract: Objective Three different doses of diethylnitrosamine (DEN)were used to establish a rat primary liver cancer ( PLC) model to establish an efficient, stable, and economical animal model of PLC. Methods Forty-five male SD rats were randomly divided into four groups: normal group, DEN 50 mg / kg dose group (low dose group),70 mg / kg dose group (medium dose group), and 200 mg / kg dose group (high dose group). There were 6 animals in the normal group and 13 animals in each of the other groups. The normal control group received no treatment. The model group and low dose groups were injected intraperitoneally twice a week during weeks 1 ~ 4 and once a week during weeks 5 ~ 12; the medium dose group was injected intraperitoneally once a week for 16 consecutive weeks; and the high dose group was administered only once in the first week. The rats in each group were then followed for 16 weeks. The establishment of the model and optimal evaluation were verified by survival rate,pathological tests, biochemical tests, liver and spleen index calculation, immunohistochemistry, enzyme-linked immunosorbent assay ( ELISA), and other assays. Results The survival rate was 100% in the normal group,46. 15% in the low dose group, 69. 23% in the medium dose group, and 84. 61% in the high dose group. The liver tissues of the rats in the normal group showed no abnormality to the naked eye; the liver of the rats in the low dose group became darker in color, rougher in surface, with a small number of cancerous nodules and slightly hard texture;the liver of the rats in the medium dose group was rough in surface, with several small cancerous nodules and scattered massive occupying nodules and hard texture; The liver of rats in the high dose group became lighter in color, slightly rougher in surface, with no obvious cancerous nodules; HE staining showed that the liver tissues of rats in the low and medium dose groups were structurally disorganized, with large cellular heterogeneity and tumor cells. HE staining showed that the liver tissues of rats in the low and medium dose groups were structurally disorganized, with large cellular heterogeneity and tumor cell formation, while the structure of the liver lobules of the high dose group was unclear, with different degrees of edema, degeneration and necrosis of liver cells, and no obvious tumor cell formation was seen. Compared with the normal group, serum liver function alanine aminotransferase ( ALT ), aspartate aminotransferase(AST), and total bilirubin(TBIL) were elevated in the low, medium, and high dose groups; ALT and AST were significantly elevated in the low dose group (P<0. 05), the difference was statistically significant,ALT, AST and TBIL were significantly elevated in the medium dose group ( P<0. 05 ), the difference was statistically significant, and the difference was statistically significant, although liver function in the high dose group was elevated, he increase was not significant, the difference was not statistically significant (P>0. 05); compared with the normal group, the international normalized ratio (INR)of coagulation function was significantly higher in the low dose group, with a statistically significant difference (P<0. 05), and the activated partial thromboplastin time (APTT), prothrombin time ( PT), and alpha-fetoprotein ( AFP ) levels were increased ( P<0. 05), and the difference was not statistically significant; serum APTT, PT, INR, and AFP levels were significantly increased in the medium dose group (P<0. 05), and the difference was statistically significant; serum PT and AFP levels were increased in the high dose group (P<0. 05), the difference was statistically significant, and plasma APTT levels were slightly increased (P>0. 05), the difference was not statistically significant; liver and spleen indexes were increased in the medium dose group (P<0. 05), the spleen index increased in the low dose group (P<0. 05), and the liver index increased in the high dose group (P<0. 05), the difference was statistically significant; the optical density value of liver tissue AFP increased significantly in the low, medium and high dose groups (P<0. 05), the difference was statistically significant. Conclusions Both the low and medium dose groups could successfully induce the PLC rat model, but the pathological changes and biochemical findings of the medium dose group were more consistent with the pathogenesis of human liver tissue from liver injury to hepatic fibrosis to cirrhosis to hepatocellular carcinoma, and the number of administrations of the drug is less, and the survival rate of the rats is higher so that a more cost-effective and superior PLC model can be established.

Abstract: Objective To investigate the patterns of syndrome changes and their possible material basis in mouse models of chronic psychological stress during the modeling process. Methods A chronic unpredictable mild stress (CUMS) method was employed to prepare a model of chronic psychological stress in mice. After 2 weeks of modeling, Xiaoyao powder (XYS group) and Sini powder ( SNS group) were administered concurrently with the modeling process. General conditions, behavioral, and biochemical indicators were compared between the modeling mice (M4 and M6 group) and the mice treated for 2 weeks (S4 and X4 group) and 4 weeks (S6 and X6 group) after chronic psychological stress. Results Compared with M4 group mice, body mass, total distance and central distance in the open field test, serum NE concentration, and gastric Ghrelin expression were increased in S4 group and X4 group mice, and sugar water preference rate and serum D-xylose concentration were increased in S4 group mice. Compared with X4 group mice, total distance in the open field test and serum D-xylose concentration were increased in the S4 group mice. Compared with M6 group mice, body mass, serum NE concentration, serum D-xylose concentration, and gastric Ghrelin expression were increased in group S6 and X6 group mice; sugar water preference rate, total distance in the open field test, and active avoidance counts in the shuttle box test were increased in X6 group mice, while their serum CORT concentration was decreased. Compared with S6 group mice, body mass, total distance in the open field test, serum D-xylose concentration, and gastric Ghrelin expression in X6 group mice were increased. Conclusions During the process of modeling chronic psychological stress in mice, the traditional Chinese medicine syndrome evolved from liver qi stagnation to liver qi stagnation with spleen deficiency. The biological bases of the syndrome changes may be the combined changes in serum CORT and NE concentrations and the decrease in gastric Ghrelin levels.

Abstract: Objective To explore the impact of peptidylarginine deiminase 2 ( Padi2 )-knockout on depressive-like behaviors in socially isolated mice. Methods Using CRISPR/ Cas9 technology, a Padi2-knockout (Padi2^{- / -}) mouse model with a C57BL / 6J background was established, and the effect of Padi2 knockout was identified by genotyping and RT-qPCR detection. Six-week-old male Padi2^{- / -} mice and wild-type C57BL / 6J mice were selected and divided into normal rearing and social isolation groups, with 15 mice per group. The normal rearing group mice were housed with 5 mice per cage, and the social isolation group was housed with 1 mouse per cage, and weighed once a week. After 4 weeks, forced swimming and open field tests were conducted. After the behavioral experiments, brain tissues were taken from mice in each group, and changes in microglia in the brains were detected by immunofluorescence. Results We successfully established Padi2^{- / -} mice. There was no difference in behavior between Padi2^{- / -}mice and C57BL / 6J mice in the normal rearing group. After social isolation, compared with C57BL / 6J mice, Padi2^{- / -}mice showed a significant increase in depressive symptoms, obvious weight gain, and a significant increase in the number of microglia in brain tissue. Conclusions Padi2 knockout exacerbated depressivelike behaviors and obesity in socially isolated mice, indicating that Padi2 is involved in the progression of depressionand may be an effective target for the prevention and treatment of depression.

Abstract: Objective To investigate the characteristics of liver injury in the Lieber-DeCarli alcoholic liver disease (ALD) mouse model and to analyze its transcriptomic profile. Methods Eighteen male C57BL / 6J mice were randomly divided into an alcohol-fed group (n= 10) and a control group (n= 8). The alcohol-fed group received a Lieber-DeCarli ethanol diet, starting with an adaptive one-week phase using incremental concentrations of ethanol (10~ 57. 3 mL / L), followed by 2 weeks of a 57. 3 mL / L concentration of 95% ethanol, for a total of 3 weeks. The control group was provided with an isocaloric control diet for 3 weeks. At the end of the study, mice were sacrificed, and serum and liver tissue samples were collected. Serum liver function markers (ALT, AST), hepatic lipids (TC,TG), reduced glutathione ( GSH), total superoxide dismutase ( T-SOD), and malondialdehyde ( MDA) were measured using biochemical assays. The levels of inflammatory cytokines ( IL-6, IL-10, TNF-α, TGF-β1) in liver tissue were assessed by ELISA. Histopathological changes in l iver tissue were examined using hematoxylin-eosin (HE) and Oil Red O staining. Immunohistochemical staining using the F4 / 80 antibody was employed to assess changes in macrophage expression. RNA-seq analysis was conducted to identify differentially expressed genes between the two groups of liver tissues, followed by GO and KEGG pathway enrichment analysis. qRT-PCR was used to validate the expression of these differentially expressed genes. Results Compared with the control group, the alcohol-fed mice exhibited a significant decrease in body weight ( P<0. 01). Serum ALT and AST levels were significantly elevated (P<0. 01), while liver tissue levels of TC, TG, and MDA were significantly increased (P<0. 05). Conversely, GSH and T-SOD levels were significantly reduced ( P<0. 05). The levels of inflammatory factors IL-6, TNF-α, and TGF-β1 were increased, which was consistent with the qRT-PCR validation Results (P<0. 05). Histological examination revealed disrupted hepatic lobular structure, with macrovesicular steatosis,microvesicular steatosis, and ballooning degeneration. Additionally, fat droplets in liver tissue were significantly increased, and macrophage expression was upregulated. Differential gene expression analysis, using a threshold of ?? log₂ FC ?? > 1 and q < 0. 05, identified 2063 differentially expressed genes, of which 1236 were upregulated and 827 downregulated. Enriched pathways included xenobiotic metabolism via cytochrome P450, cytokine-cytokine receptor interaction, chemokine signaling, steroid hormone biosynthesis, glutathione metabolism, and retinol metabolism. ( P<0. 05). qRT-PCR validation confirmed the significant upregulation ( e. g. , Mmp12, Gstm3,Cyp2a22) and downregulation (e. g. , Serpina1e, Acmsd, Mup3d) of 10 genes from each category, consistent with the transcriptome sequencing Results. Conclusions The primary pathological mechanisms underlying alcoholic liverinjury involve pathways related to xenobiotic metabolism and act via cytochrome P450, cytokine-cytokine receptor interaction, chemokine signaling, glutathione metabolism, and retinol metabolism.

Abstract: Objective In this study, 2, 4-dinitrochlorobenzene ( DNCB ) was used to induce the establishment of an atopic dermatitis (AD) model in BALB/ c homozygous mice to simulate the skin inflammatory complications in patients with clinical malignancies. Methods BALB/ c mice were divided into different groups:negative control group ( NC group), model group ( MODEL group), atopic dermatitis group ( AD group), and dexamethasone group (DEX group). After the mice in MODEL group and DEX group were inoculated with S-180 tumor cells in the axilla, MODEL group, AD group and DEX group were stimulated with DNCB on the dorsal skin and the ear to establish an animal model of atopic dermatitis in tumor-bearing mice. Changes in body weight were observed and recorded, the dorsal skin condition of mice was assessed after the last administration of the drug, the spleen was taken to calculate the spleen coefficient, the difference in the mass of mouse ear slices was determined to calculate the degree of auricular swelling and the rate of inhibition of swelling, and histopathological tests were performed on the dorsal skin tissues to detect the levels of IgE, TNF-α, IL-4, and IL-17 in the serum using an ELISA assay. Results Compared with the NC group, the skin of mice in the MODEL and AD groups showed erythematous, papular, scaly and mossy changes, accompanied by weight loss, and a significant increase in splenic coefficient and auricular swelling. Pathologic findings showed an incomplete skin structure, a significant increase in skin thickness, a large infiltration of inflammatory cells, and an increase in the number of mast cells. Serum levels of IgE, TNF-α, IL-4 and IL-17 were increased. Compared with the MODEL group, the DEX group showed an improvement in all the assays. Conclusions DNCB excitation can successfully establish an animal model of AD in hormonal mice, which is drugcontrollable, which provides a useful scientific tool for conducting scientific research related to malignant tumors and skin inflammation.

Abstract: Objective To establish, based on new diagnostic criteria for osteoradionecrosis of the jaws (ORNJ), an animal model with a controllable bone defect range and early and mild ORNJ characteristics. Methods A method involving single high-dose irradiation with an electron linear accelerator was applied. According to the irradiation dose, the animals were divided into a group (control group); b group (12 Gy group); and c group (14 Gy group). Six rabbits were randomly assigned to each group, b and c groups were uniformly selected for further radiotherapy of the right mandibular area. After 1 week, all animals were prepared with a standard bone defect at the right mandibular angle. After 4 weeks, the animals were euthanized, and general observations were conducted. CBCT scans and HU values of mandibular specimens, HE staining histological observations, Trap staining histology, and osteoclast counts were compared. And the statistical analysis were carried out. Results 4 weeks after the bone defect was created, the soft tissue in the surgical area of b and c groups showed mild swelling, purple skin, and erosion and ulceration of the oral mucosa, with c group showing the most significant changes. CBCT examination showed that the cortical bone of the irradiated areas in b and c groups was rough, while that of c group had poor continuity. No significant changes were observed in the extent of the bone defect in any group,b and c groups had significantly decreased HU values compared with a group. HE staining and histological observation suggested that the bone continuity of b and c groups had deteriorated; there was a large amount of fibrotic tissue proliferation, an increase in blank bone pits without cells, and an infiltration of inflammatory cells. Trap staining revealed a significant increase in osteoclasts in c group compared with a group. Conclusions By using an electron linear accelerator, a single dose of 14 Gy irradiation was applied to the mandibular angle area to create bone defects with consistent specifications in rabbits. After 4 weeks, an animal model that met the early diagnostic criteria of ORNJ and had controllable consistency in the range and degree of bone defects was established.

Abstract: Objective Compare the depression phenotypes of a breast cancer tumor-bearing mouse model constructed using two different method and a mouse model of breast cancer depression with clinical manifestations, as well as assess their suitability for basic research. Methods We constructed a tumor model with 4T1 breast cancer cells alone (4T1 group)and a tumor-depression composite model given chronic unpredictable mild (CUMS) (4T1 +CUMS group). The experimental period was 42 d, and the body mass, tumor volume, and survival time of the mice were monitored throughout the whole process. Two depressive behavioral tests (of sucrose preference test, open field test, tail suspension test, and elevated plus maze ) were performed on the 15th and 29th days, respectively. Hematoxylin-eosin (HE)staining was used to observe the pathological changes of hippocampal neurons in brain tissue sections. Results (1)Body mass: The body mass of the 4T1 group and 4T1 + CUMS group began to decrease from29 d, and the body mass of the 4T1 + CUMS group was significantly lower than that of the 4T1 group and Control group at the end of the experiment (P<0. 001). (2) Tumor volume: There was no significant difference in the growth rate of tumors between the two model groups throughout the experiment (P>0. 05). (3)Survival time: The survival rates of the 4T1 group and 4T1 + CUMS group were 100% and 60%, and the first death of mice in the 4T1 + CUMS group was on the 36th day. (4) Behavior test of depression: There was no significant difference between the three groups in the first depressive behavior tests (P>0. 05), and the two groups showed obvious depressive phenotypes in the second behavioral tests. The sucrose preference index and activity distance in the central area were significantly decreased in the two model groups (P<0. 001), and the immobility time was significantly increased (P<0. 001).(5)Pathological section of brain tissue: On pathological examination of brain tissue, we observed a reduced number of neuronal cells in the hippocampus of the 4T1 group and 4T1 + CUMS group, their morphology was irregular, the arrangement between the cells was disordered and the gap was unclear, and some nucleoli were blurred. Conclusions Although the tumor-only method and the tumor with compound stress stimulation method can both be used to prepare breast cancer depression models, the tumor-only modeling method is simpler and the mortality rate after successful modeling is higher. The long window of time is convenient for subsequent drug administration and detection, and the causes of the depression phenotype are more in line with the clinical causes and manifestations. Therefore, the 4T1 model can provide a reference model for future animal experiments on breast cancer tumor-related depression.

Abstract: Objective To explore the function of electroacupuncture ( EA) on body mass, fasting blood glucose, heat pain threshold, and transient receptor potential vanilloid 4 (TRPV4) in the dorsal root ganglia (DRG) of rats with diabetic neuropathic pain (DNP). Methods A DNP rat model was formed by intraperitoneally injecting the animals with STZ. From days 15 to 21, bilateral Zusanli and Kunlun points of the DNP rat model were treated with electroacupuncture once daily for 30 min. We then measured their body mass, fasting blood glucose, and heat pain threshold. The co-expression of TRPV4 and NeuN in the rat L4 ~ L6 DRG was detected by immunofluorescence. The effects of the TRPV4 agonist GSK1016790A on body mass, fasting blood glucose, and the heat pain threshold of DNP rats treated with electroacupuncture were detected. Results After the 7th day, body mass was significantly decreased (P<0. 01) and fasting glucose was significantly increased (P<0. 01) in the model group compared with the normal group. After the 21st day, compared with the model group, heat pain threshold of the model + electroacupuncture group was significantly higher (P<0. 01); the Results of co-expression of TRPV4 and NeuN immunofluorescence on rat L4 ~ L6 DRG showed that: the expression of positive cells in the model group was significantly higher (P<0. 01) than that in the normal group, the co-expression of TRPV4 and NeuN positive cells in L4 ~ L6 DRG of rats in the model + electroacupuncture group was significantly lower ( P<0. 01) than that in the model group. The TRPV4 agonist GSK1016790A can reverse the downregulation of thermal pain threshold induced by electroacupuncture in DNP rats (P<0. 01). Conclusions Electroacupuncture alleviated the DNP induced by STZ, and its mechanism may involve the inhibition of TRPV4 protein expression in the DRG.

Abstract:Depression is a common chronic mental illness. Animal models of depression are widely used to study the pathogenesis of depression as well as in the development of new antidepressant drugs. The consistency and reliability of animal models of depression to simulate human depressive symptoms directly affect the study result.However, at present, no animal models have been found that are completely consistent with the onset of human depression, which hinders the in-depth study of depression. By using scientific and reasonable modeling and evaluation method, the pathological state of depression can still be simulated to a large extent, providing a basis for further in-depth research of the pathogenesis of depression and the development of effective antidepressant drugs. This paper aims to provide a reference for researchers by reviewing several commonly used animal models and behavioral tests of depression.

Abstract:Fibrosis is a pathological process characterized by an increase in connective tissue and a decrease in parenchymal cells within organ tissues. During its progression, fibrosis can lead to structural damage and functional decline of the affected organ. In recent years, it has found that non-selective cation channel transient receptor potential vanilloid subfamily 1 (TRPV1) channel is closely related to fibrosis. When this channel is activated, it can increase the intracellular cation concentration and cause corresponding physiological and pathological changes, playing a particularly crucial role in regulating tissue fibrosis. Animal models have become important tools in studies into the mechanism by which the TRPV1 channel induces organ fibrosis. This article reviews the role of TRPV1 channels in the fibrosis of organs such as the heart, kidney, and pancreas in mice. The relevant signaling pathways in which TRPV1 channels participate to regulate fibrosis are summarized to provide new ideas for studying the pathogenesis of fibrosis and the development of targeted drugs.

Abstract:Depression is a common mental illness that has a negative impact on patient quality of life, as well as representing a serious economic burden to society. The strategy of interfering with depression based on signaling pathways has recently attracted widespread attention. The cAMP pathway is an important cellular signaling pathway that plays a key role in the pathogenesis of and interventions for depression. In this review, we addresses recent studies on cAMP pathway-based interventions in depression, to provide a theoretical basis for regulating the cAMP pathway and its cascade mechanism in depression

Abstract:Japanese encephalitis virus ( JEV) usually evades the inhibitory effect of the innate immunity factor type Ⅰ interferon (Ⅰ-IFN) when it infects human cells and tissues. The virus then causes a series of serious symptoms, such as spasticity, neurodegenerative lesions, neuroinflammation, and even death. Generally, JEVescapes innate immunity by inhibiting IFN-α/ β production and the interferon Janus kinase-signal transducer and activator of transcription signaling pathway. Because of this special immune escape mechanism, various mouse infection models have been constructed for the study of the pathogenesis of and therapeutic regimens for JEV infections. In this review, based on an exposition of the IFN immune escape mechanism of JEV, we systematically introduce the concept of JEV-infected mouse models and analyze the characteristics of these models and the degree to which they simulate human symptoms. The intention is to develop various new JEV-infected mouse models based on potential new research targets and provide novel ideas for animal models for JEV research.

Abstract:Japanese encephalitis virus ( JEV) usually evades the inhibitory effect of the innate immunity factor type Ⅰ interferon (Ⅰ-IFN) when it infects human cells and tissues. The virus then causes a series of serious symptoms, such as spasticity, neurodegenerative lesions, neuroinflammation, and even death. Generally, JEVescapes innate immunity by inhibiting IFN-α/ β production and the interferon Janus kinase-signal transducer and activator of transcription signaling pathway. Because of this special immune escape mechanism, various mouse infection models have been constructed for the study of the pathogenesis of and therapeutic regimens for JEV infections. In this review, based on an exposition of the IFN immune escape mechanism of JEV, we systematically introduce the concept of JEV-infected mouse models and analyze the characteristics of these models and the degree to which they simulate human symptoms. The intention is to develop various new JEV-infected mouse models based on potential new research targets and provide novel ideas for animal models for JEV research.

Abstract:Alzheimer’s disease (AD) is a common neurodegenerative disease. Traditional Chinese medicine is effective in treating AD. The combination of disease and syndrome with animal model is the basis and premise of related research. In this paper, studies on non-transgenic AD animal models and AD disease and syndrome combined animal models are summarized. It was found that, there are eight common non-transgenic AD animal models,including aging type (natural aging type, rapid aging type and induced aging type) and injection-induced injury type (Aβ injection animal model, Tau damage type, damage model of cholinergic system, neuroinflammatory model and aluminum poisoning induction model). AD disease and syndrome combined with animal models include kidney deficiency / kidney deficiency and essence deficiency / kidney deficiency and pulp emptying, phlegm turbidity blocking orifice, blood stasis blocking collaterals, phlegm stasis interlocking, liver depression and phlegm fluid stagnation heat. In this paper, the advantages and disadvantages of each model replication method are reviewed, in order to provide reference and support for the future study of AD disease and syndrome combined with animal models.