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Objective　To investigate the protective effect of Genistein against nonalcoholic fatty liver disease （NAFLD） in ovariectomized （OVX） mice and its mechanism. Methods　A total of 40 female C57BL/6 mice, aged 6 weeks, were used to establish an OVX mouse model, and then they were randomly divided into blank group, 4-week model group, 6-week model group, 8-week model group, and 10-week model group, with 8 mice in each group. Under the same environmental conditions, the mice were given high-fat diet for modeling, and pathological examination showed that NAFLD was successfully induced by 10-week high-fat diet. Another 40 female C57BL/6 mice, aged 6 weeks, were randomly divided into blank group, sham operation group （Sham group), OVX group, OVX+L-Genistein （4 mg/kg body weight） group, and OVX+H-Genistein （8 mg/kg body weight） group. The mice in the Sham group were given the same procedure of OVX, without the ligation of the ovarian artery and the resection of the ovary. The mice in the blank group were given normal diet, and those in the other groups were given high-fat diet. Genistein was dissolved in DMSO, and the mice in the Sham group and the OVX group were treated with solvent solution alone by gavage, once a day for 10 consecutive weeks. Body weight and visceral index were recorded, and the mice were sacrificed to collect serum and liver tissue. Kits were used to measure the activity of alanine aminotransferase （ALT） and aspartate aminotransferase （AST） and the serum levels of triglyceride （TG） and total cholesterol （TC), and HE staining and oil red O staining were used to observe liver histopathology； Western blot was used to measure the protein expression levels of sterol regulatory element-binding protein 1c （SREBP-1c） and peroxisome proliferator-activated receptor alpha （PPARα） associated with lipid metabolism in liver tissue. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the Dunnett-t test was used for further comparison between two groups. Results　After 10 weeks of high-fat diet, the OVX+L-Genistein group and the OVX+H-Genistein group had significantly lower body weight, liver index, and liver tissue weight （all P<0.05). In addition, Genistein significantly downregulated the serum levels of TC and TG （P<0.05） and reduced the activities of serum AST and ALT （P< 0.05). HE and oil red O staining showed that compared with the OVX group, the OVX+L-Genistein group and the OVX+HGenistein group had a significant reduction in the accumulation of lipid droplets. Western blot showed that after Genistein intervention, there was a significant reduction in the protein expression level of SREBP-1c and a significant increase in the protein expression level of PPARα （P<0.05). Conclusion　Genistein exerts a protective effect against NAFLD in OVX mice possibly by regulating the expression of SREBP-1c and PPARα, thereby promoting fatty acid oxidation and inhibiting liver lipid synthesis. [ABSTRACT FROM AUTHOR]

Objective　To investigate the mechanism of action of Xiayuxue decoction in inhibiting nonalcoholic fatty liver disease （NAFLD） induced by high-fat diet in mice by regulating nucleotide binding oligomerization domain like receptor containing pyrin domain protein 6 （NLRP6). Methods　A total of 15 male C57BL/6 mice were randomly divided into low-fat diet （LFD） group, high-fat diet （HFD） group, and Xiayuxue decoction-HFD group （XYXD group）, with 5 mice in each group. Liver function parameters （alanine aminotransferase ［ALT］ and aspartate aminotransferase ［AST］） and blood lipid metabolic indicators （triglycerides ［TG］ and total cholesterol ［TC］） were measured； HE staining and oil red O staining were performed for liver tissue to observe histomorpholoty and lipid droplet deposition； quantitative real-time PCR was used to measure the expression levels of inflammatory factors （tumor necrosis factor-α ［TNF-α］, interleukin-1β ［IL-1β］, interleukin-18 ［IL-18］, and NLRP6） in liver tissue； Western blot was used to measure the protein expression levels of NLRP6, nuclear factor-kappa B （NF-κB）, and NF-κB p65； immunohistochemistry was used to measure the expression of NLRP6 and CD68. Mouse Raw264.7 cells were treated with palmitic acid （PA）, lipopolysaccharide, and serum containing Xiayuxue decoction to observe inflammation. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for further comparison between two groups. Results　Compared with the LFD group, the HFD group had significant increases in the serum levels of ALT, AST, TC, and TG （all P<0.05). Liver histopathological examination showed that the HFD group had marked hepatic steatosis and a signficant increase in NAS score （P<0.05）, and quantitative real-time PCR showed significant increases in the inflammatory factors such as IL1β and IL-18 and a significant reduction in the expression of NLRP6 （all P<0.05). Immunohistochemistry showed that the expression of NLRP6 showed a similar trend as that of the macrophage marker CD68. Western blot showed that after the downregulation of NLRP6 expression, there was a significant increase in phosphorylated NF- κB p65 （P< 0.05). Compared with the HFD group, Xiayuxue decoction effectively improved liver inflammation, upregulated the expression of NLRP6, and downregulated phosphorylated NF- κB p65 in HFD mice （all P<0.05). After Raw264.7 cells were treated with PA, NLRP6 was downregulated to promote the progression of inflammation （P<0.05）, and treatment with Xiayuxue decoction could upregulate NLRP6 and inhibit inflammation NF- κB （P<0.05). Conclusion　Xiayuxue decoction can effectively improve hepatic steatosis and liver inflammation in a mouse model of NAFLD, possibly by regulating NLRP6/NF-κB to alleviate macrophage activation. [ABSTRACT FROM AUTHOR]

Objective To investigate the role and mechanism of action of Yinchenhao Decoction in inhibiting ferroptosis of hepatocytes in mice with autoimmune hepatitis. Methods A total of 18 specific pathogen-free female C57BL/6 mice were selected and divided into normal group, model group, and treatment group using a random number table, with 6 mice in each group. The mice in the model group and the treatment group were injected with concanavalin A (Con A) via the caudal vein to establish a mouse model of autoimmune hepatitis, and those in the normal group were injected with normal saline. The mice in the treatment group were given prophylactic treatment with Yinchenhao Decoction (4.68 g crude drug/kg) by gavage at 14 days before modeling, and Con A was injected after the last gavage. The levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), interferon gamma (IFN-γ), tumor necrosis factor-α (TNF-α), iron ion, glutathione (GSH), reactive oxygen species (ROS), adenosine triphosphate (ATP), and malondialdehyde (MDA) were measured； liver index and spleen index were calculated； the expression levels of GPX4 and SLC7A11 were measured； liver histopathological changes were compared between groups. A oneway analysis of variance was used for comparison of normally distributed continuous data between three groups, and the least significant difference t-test was used for further comparison between two groups. Results Compared with the normal group, the model group had significant increases in liver index, spleen index, ALT, AST, IFN-γ, TNF-α, iron ion, ROS and MDA (all P< 0.05) and significant reductions in the content of GSH and ATP and the protein expression levels of GPX4 and SLC7A11 (all P< 0.05). Compared with the model group, the treatment group had significant reductions in liver index, spleen index, ALT, AST, IFN-γ, TNF-α, iron ion, ROS and MDA (all P<0.05) and significant increases in the content of GSH and ATP and the protein expression levels of GPX4 and SLC7A11 (all P<0.05). HE staining showed that compared with the normal group, the model group showed massive hepatocyte degeneration and necrosis and inflammatory cell aggregation at the portal area, and compared with the model group, the treatment group had alleviation of liver necrosis and inflammatory infiltration. Conclusion Liver injury induced by Con A may be associated with ferroptosis. Yinchenhao Decoction can increase the protein expression levels of SLC7A11 and GPX4 protein and thus inhibit ferroptosis of hepatocytes induced by Con A. [ABSTRACT FROM AUTHOR]

Objective To investigate the therapeutic effect of Qingjie Huagong decoction (QJHGD) on a mouse model of severe acute pancreatitis (SAP) and the mechanism of action of QJHGD against inflammatory response. Methods A total of 36 male C57BL/6J mice were randomly divided into blank group, model group, Western medicine group (ulinastatin), and low-, middle-, and high-dose QJHGD groups, with 6 mice in each group. All mice except those in the blank group were given 5% sodium taurocholate by retrograde pancreaticobiliary injection to establish a model of SAP. After modeling, the mice in the low-, middle-, and high-dose groups were given QJHGD (1，2, and 4 g/kg, respectively) by gavage, and those in the Western medicine group were given intraperitoneal injection of ulinastatin (5×104 U/kg), for 7 days in total. HE staining was used to observe the histopathological changes of the pancreas； ELISA was used to measure the levels of α-amylase, lipase, interleukin-1β（IL-1β), interleukin-6（IL-6), interleukin-8（IL-8), interleukin-18（IL-18), and tumor necrosis factor-α（TNF-α) in mice； RT-qPCR was used to measure the mRNA expression levels of NOD-like receptor protein3 (NLRP3), Toll-like receptor 4（TLR4), and nuclear factor-kappa B (NF-κB) in pancreatic tissue； immunohistochemistry was used to measure the positive expression rates of NLRP3, TLR4, and NF-κB in pancreatic tissue； Western blot was used to measure the protein expression levels of NLRP3, TLR4, NF-κB, IL-1β, and IL-6. An analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for further comparison between two groups. Results Compared with the blank group, the model group had diffuse destruction of pancreatic tissue structure, focal dilatation of pancreatic lobular septum, pancreatic acinar atrophy, and massive inflammatory cell infiltration, as well as significant increases in the content of α-amylase, lipase, IL-1β, IL-6, IL-8, IL-18, and TNF-α（all P<0.05), the mRNA expression levels and positive expression rates of NLRP3, TLR4, and NF-κB (all P< 0.05), and the protein expression levels of NLRP3, TLR4, NF-κB, IL-1β, and IL-6（all P<0.05) . Compared with the model group, the low-, middle-, and high-dose QJHGD groups and the Western medicine group had slightly tighter and more intact structure of pancreatic tissue, ordered arrangement of pancreatic acinar cells, a small amount of inflammatory cell infiltration, and hemorrhagic foci of pancreatic lobules, as well as significant reductions in the content of α-amylase, lipase, IL-1β, IL-6, IL-8, IL-18, and TNF-α（all P<0.05), the mRNA expression levels and positive expression rates of NLRP3, TLR4, and NF-κB (all P<0.05), and the protein expression levels of NLRP3, TLR4, NF-κB, IL-1β, and IL-6（all P<0.05) . Conclusion QJHGD may exert a protective effect on the pancreatic tissue of SAP mice by inhibiting the activation of NLRP3/TLR4/NF-κB signaling pathway-related proteins, reducing the release of inflammatory mediators, and preventing the enhancement of inflammatory cascade response. [ABSTRACT FROM AUTHOR]

Objective To investigate the intervention mechanism of Dendrobium officinale leaf fermentation fluid in mice with alcoholic hepatitis. Methods A total of 70 healthy male C57BL/6J mice, aged 6-8 weeks, were randomly divided into normal group, model group, liquid feed control group, silybin group, and low-, middle-, and high-dose Dendrobium officinale leaf fermentation fluid groups, with 10 mice in each group. The mice in the normal group were given normal diet, and those in the other groups were given Lieber-DeCarli classic liquid diet for 8 weeks to induce alcoholic hepatitis. During modeling, the mice in the low-, middle-, and high-dose Dendrobium officinale leaf fermentation fluid groups were given Dendrobium liquid manufactured by Warmen Pharmaceutical, and the mice in all the other groups were given pure water； the mice in the normal group, the model group, and the liquid feed control group were given normal saline by gavage, those in the silybin group were given silybin 0.25 mL/10 g by gavage, and those in the low-, middle-, and high-dose Dendrobium officinale leaf fermentation fluid groups were given Dendrobium officinale leaf fermentation fluid at a dose of 0.125 mL/10 g，0.250 mL/10 g, and 0.375 mL/10 g, respectively, by gavage, once a day. At week 8, chloral hydrate was injected intraperitoneally for anesthesia, and blood samples were collected from the eyeball. After serum was separated, the biochemical method was used to measure the levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) ； HE staining and oil red staining were used to observe liver histopathology and lipid accumulation in mice； multiplex Luminex assay was used to measure the serum levels of interleukin-6 (IL-6), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), and CCL2； quantitative real-time PCR, Western blot, and immunofluorescence assay were used to measure the protein expression levels of NLRP3, caspase-1, caspase-11, gasdermin D (GSDMD), N-terminal gasdermin D (GSDMD-N) in liver tissue. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for further comparison between two groups. Results Compared with the normal group, the model group had significant increases in the serum levels of AST, ALT, IL-6, IL-1β, TNF- α, and CCL2 (all P<0.05), and compared with the model group, the high-dose Dendrobium officinale leaf fermentation fluid group had significant reductions in the serum levels of AST, ALT, IL-6, IL-1β, TNF-α, and CCL2 (all P<0.05) . HE staining showed that the model group had disordered structure of hepatic lobules, with a large number of steatosis vacuoles and massive cell necrosis, and compared with the model group, the high-dose Dendrobium officinale leaf fermentation fluid group had alleviation of liver histopathological injury, intact structure of most hepatic lobules, and a small amount of inflammatory cell infiltration. Oil red staining showed that the model group had accumulation of large and small lipid droplets in the liver and a significant increase in liver fat content, and compared with the model group, the high-dose Dendrobium officinale leaf fermentation fluid group had significant alleviation of hepatic steatosis, with the presence of sporadic small lipid droplets. Immunofluorescence assay of liver tissue showed that compared with the normal group, the model group had a significant increase in the ratio of GSDMD-positive staining area in hepatocyte cytoplasm (P<0.001), and compared with the model group, the high-dose Dendrobium officinale leaf fermentation fluid group had a significant reduction in such ratio in hepatocyte cytoplasm (P<0.001) . Quantitative real-time PCR showed that compared with the normal group, the model group had significant increases in the protein expression levels of NLRP3, caspase-1, caspase-11, GSDMD, GSDMD-N, interleukin-18 (IL-18), and IL-1β in liver tissue (all P<0.05), and compared with the model group, the high-dose Dendrobium officinale leaf fermentation fluid group had significant reductions in the protein expression levels of NLRP3, caspase-1, caspase-11, GSDMD, GSDMD-N, IL-18, and IL-1 (all P<0.05) . Compared with the model group, the high-dose Dendrobium officinale leaf fermentation fluid group had significant reductions in the protein expression levels of caspase-1 and caspase-11 (both P<0.05), with a relative expression level of caspase-1 of 1.757 (reduced by 26.6% compared with the model group) and a relative expression level of caspase-11 of 0.455 (reduced by 70.3% compared with the model group), suggesting that caspase-11 showed a greater reduction than caspase-1. Conclusion Dendrobium officinale leaf fermentation fluid can alleviate alcoholic hepatitis in mice, possibly by inhibiting the non-classical cell pyroptosis pathway mediated by caspase-11. [ABSTRACT FROM AUTHOR]

Objective To investigate the effect of Xuefu Zhuyu decoction on nonalcoholic fatty liver disease (NAFLD) and its material basis. Methods In experiment 1 for exploring the effect of Xuefu Zhuyu decoction on mice with NAFLD induced by high-fat diet, 50 healthy male C57 BL/6 J mice were randomly divided into normal group, model group, high-and low-dose Xuefu Zhuyu decoction groups, and obeticholic acid control group, with 10 mice in each group. The mice in the normal group were given control diet, and those in the other groups were given high-fat diet. Gastric administration was started at week 13, and related samples were collected at the end of week 16. Food intake and body weight were recorded, enzyme-linked immunosorbent assay was used to measure the serum level of fasting insulin, fasting blood glucose was measured, and insulin resistance index was calculated. HE staining and NAFLD activity score (NAS) were used to observe liver histopathology in mice, oil red O staining was used to observe lipid deposition, and triglyceride (TG) level in liver tissue and serum alanine aminotransferase (ALT) level were measured. In experiment 2 for exploring the effect of different extracts of Xuefu Zhuyu decoction on mice with NAFLD induced by high-fat diet, the methods of water decocting, water extraction and alcohol precipitation, and petroleum ether extraction were used to obtain the extracts 1, 2, and 3 of Xuefu Zhuyu decoction, and 54 healthy male C57 BL/6 J mice were randomly divided into normal group, model group, Xuefu Zhuyu decoction extract 1, 2, and 3 groups, and Xuefu Zhuyu decoction control group, with 9 mice in each group. The mice in the normal group were given control diet, and those in the other groups were given high-fat diet. Gastric administration was started at week 13, and related samples were collected at the end of week 16. Food intake and body weight were recorded, and enzyme-linked immunosorbent assay was used to measure the serum level of fasting insulin, fasting blood glucose was measured, and insulin resistance index was calculated. HE and NAS were used to observe liver histopathology in mice, oil red O staining was used to observe lipid deposition, and the levels of TG and gamma-glutamyl transpeptidase (GGT) in liver tissue and the serum level of ALT were measured. The t-test was used for comparison of normally distributed continuous data between two groups; a one-way analysis of variance was used for comparison between multiple groups, and the least significant difference t-test was used for further comparison between two groups. The Kruskal-Wallis H test was used for comparison of non-normally distributed continuous data between multiple groups and further comparison between two groups. Results In experiment 1, compared with the model group, the high-and low-dose Xuefu Zhuyu decoction groups and the obeticholic acid control group had significant reductions in body weight, insulin resistance index, the distribution of vacuolar lipid droplets in liver tissue, intralobular inflammation, the ballooning degeneration of hepatocytes, NAS score, the level of TG in liver tissue, and the serum level of ALT (all P<0.05). Compared with obeticholic acid, high-and low-dose Xuefu Zhuyu decoction had a significantly better effect in reducing body weight, insulin resistance index, and total NAS score (all P<0.05), and low-dose Xuefu Zhuyu decoction had a significantly better effect in improving serum ALT (P<0.05). In experiment 2, compared with the model group, the Xuefu Zhuyu decoction extract 1, 2, and 3 groups had significant reductions in fasting blood glucose, insulin resistance index, the distribution of lipid droplets in liver tissue, intralobular inflammation lesions, the ballooning degeneration of hepatocytes, total NAS score, and the level of TG in the liver (all P<0.05). Compared with the model group, the extract 1 group had a significant reduction in body weight (P<0.05); the extract 2 and 3 groups had a significant reduction in the serum level of ALT (P<0.05); the extract 2 group had a significant reduction in the level of GGT in liver tissue (P<0.05). The extract 2 of Xuefu Zhuyu decoction had the closest effect to compound Xuefu Zhuyu decoction. Conclusion Xuefu Zhuyu decoction and its extracts can help to achieve varying degrees of improvement in NAFLD induced by high-fat diet in mice, and the extract 2 of Xuefu Zhuyu decoction might be the main material basis for Xuefu Zhuyu decoction. [ABSTRACT FROM AUTHOR]

Objective: Capsaicin, the pungent analgesic substance of hot peppers which produces a burning sensation and pain is known to affect Substance P and central opioid activities. This experiment was designed to test the effect of capsaicin on alcohol consumption in C57BL/6 and DBA/2 mice. These two strains are known to differ in both their alcohol consumption and their endogenous opioid distribution and response to alcohol. It is hypothesized that this effect may be mediated by both increases Substance P and decreases beta-endorphin. Methods: After i.p. administration of 0.01 and 0.001 mg/kg of capsaicin with a vehicle or the vehicle alone as the control for eight days in C57BL/6 and DBA/2 mice on limited access alcohol model, Capsaicin's effects on 2-hour alcohol, 22-hours water, 24-hours food intake and body weight were studied. Results: In this study, as expected, C57BL/6 mice drank significantly more alcohol than DBA/2 mice under baseline conditions. Capsaicin at both doses tested significantly reduced baseline alcohol consumption in C57BL/6 but not DBA/2 mice. These effects were selective for alcohol as capsaicin did not disrupt food or water consumption. Conclusion: These results demonstrate that capsaicin differentially affects those mechanisms underlying alcohol consumption in two strains of mice known to differ in their preference for and consumption of alcohol. This effect is hypothesized to be related to differences in the response of the endogenous opioid system. [ABSTRACT FROM AUTHOR]

Objective To investigate the effect and mechanism of fenofibrate (Fen) on acute lung injury (ALI). Methods (1) In vivo experiment: thirty male C57BL/6 mice were divided into 6 groups by random number table method including the normal group, the model group (LPS group), the positive drug (dexamethasone) group, and the fenofibrate (Fen) low, medium and high dose (20, 40 and 80 mg/kg) groups. After 12 h of administration, the rats were sacrificed, wet/dry ratio of lung tissue was recorded. The contents of TNF- α, IL-1β and IL-6 in lung tissue were detected by enzyme linked immunosorbent assay (ELISA). The total protein content and immune cell number in the bronchoalveolar larage fluid (BALF) were determined by BCA colorimetry and Wright-Giemsa staining. Hematoxylin-eosin (HE) staining was used to observe lung lesions and pathological scores. Western blotting was used to detect the expressions of B-cell lymphoma-2 (Bcl-2), bcl- 2-Associated X (Bax), c-Jun N-terminal kinase (JNK) and phosphorylated c-Jun N-terminal kinase (p-JNK) in lung tissues. (2) In vitro experiment: the experiment consisted of 7 groups including the control group, the LPS (10 mg/L) group, the LPS+Fen (5 μmol/L) group, the LPS+Fen (10 μmol/L) group, the LPS+Fen (20 μmol/L) group. A549 cells were treated with LPS (10 mg/L) and Fen (5, 10 and 20 μmol/L) for 12 h, respectively. DCFH-DA, Annexin V-FITC/PI and Western blot assay were used to detect effects of Fen on ROS levels, apoptosis and expression of apoptosis-related proteins Bcl-2, Bax and p-JNK in A549 cells. After LPS (10 mg/L) +Fen (20 μmol/L) treatment, H2O2 (100 μmol/L) was added to observe the changes of ROS, apoptosis rate and expression of apoptosis-related proteins Bcl-2 and Bax. In addition, LPS (10 mg/L) and Fen (20 μmol/L) were treated and JNK agonist anisomycin (3 μmol/L) was added to observe the expression changes of Bcl-2 and Bax. Results (1) Compared with the normal group, the lung wet-dry ratio, lung histopathological score, the contents of TNF- α, IL-1β, IL-6, the number of total proteins and immune cells in alveolar lavage fluid, p-JNK and Bax expression were significantly increased in the model group, and the expression of Bcl-2 was significantly decreased. Compared with the model group, the changes of above indexes were significantly reversed in the Fen low, medium and high dose groups (20, 40, 80 mg/kg). (2) Fenofibrate significantly reduced ROS content, decreased apoptosis rate, inhibited p-JNK and Bax expression and promoted Bcl-2 expression in LPS-induced A549 cells, which were reversed by H2O2 treatment. Up-regulation of Bcl-2 and down-regulation of Bax expression induced by Fen in LPS-induced A549 cells were reversed by anisomycin treatment. Conclusion Fenofibrate inhibits apoptosis through ROS/JNK signaling, so as to reduce ALI. [ABSTRACT FROM AUTHOR]

Objective To explore the protective effect of exosomes derived from M2 macrophages (M2 Exos) on concanavalin (Con) A induced autoimmune hepatitis in mice. Methods RAW264.7 macrophages were treated with IL-4 (20 μg/L) for 24 h to obtain cell culture supernatant containing M2 Exos. M2 Exos were separated by ultracentrifugation and identified by transmission electron microscope, nanoparticle tracking analysis and Western blot assay. Immunofluorescence was used to detect whether M2 Exos were uptaken by RAW264.7 macrophages. Twelve C57BL/6J mice were randomly divided into PBS group, DiR-M0 Exos group, DiR-M2 Exos group and DiR dye control group. PBS solution, DiR-M0 Exos (100 μg), DiR-M2 Exos (100 μg) and DiR dye were given to corresponding groups through tail veins after the injection of Con A (15 mg/kg) for 1h. The distribution of Exos in liver, spleen, heart, lung, kidney and intestine of mice with autoimmune hepatitis was observed by vivo imaging system. Twenty C57BL/6J mice were randomly divided into control group (PBS solution), Con A group (15 mg/kg Con A), M0 Exos group (15 mg/kg Con A + 200 μg M0 Exos) and M2 Exos group (15 mg/kg Con A+200 μg M0 Exos), 5 mice in each group. Peripheral blood and liver tissues were collected 12 h after Con A injection. The serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were determined by automated biochemistry analyzer. The histological profiles of liver tissue were observed by hematoxylin-eosin (HE) staining. The mRNA expression levels of liver TNF-α and IL-6 were detected with quantitative real-time PCR (qPCR). The changes of liver macrophage subpopulations were detected with flow cytometric analysis. Results M2 Exos were successfully induced and isolated, which can be taken up by RAW264.7 macrophages in large quantities. M2 Exos were mainly absorbed by liver and spleen of mice through tail vein injection. Compared with Control group, the serum levels of ALT and AST were significantly increased in Con A group (both P＜0.05). The disordered liver structure and increased necrosis areas were found in Con A group. Meanwhile, the mRNA expression of TNF- α and IL-6 in liver increased (both P＜0.05). The infiltration of liver monocyte-derived macrophages (MoMFs) increased (P＜0.05). Compared with Con A group, the serum levels of ALT and AST were significantly decreased in M2 Exos group (both P＜0.05). M2 Exos group also showed lower liver TNF-α, IL-6 mRNA expression levels (both P＜0.05) and lower MoMFs infiltration. HE staining showed that the necrosis areas of liver cells were relieved in M2 Exos group. Conclusion M2 Exos plays a protective role in autoimmune hepatitis in model mice，and its mechanism may be related to the decreased production of liver inflammatory cytokines and decreased recruitment of MoMFs. [ABSTRACT FROM AUTHOR]

Objective To investigate the protective effect of fecal microbiota transplantation (FMT) on mice with acute - on - chronic liver failure (ACLF) and its effect on intestinal flora. Methods A total of 40 mice were randomly divided into control group (CON group), model group (MOD group), FMT group (feces of the mice in the CON group were used as fecal microbiota donor), and FMT model group (ANFMT group, with feces of the mice in the MOD group as fecal microbiota donor), with 10 mice in each group. All mice were observed in terms of body weight, death, liver histopathology, and changes in aspartate aminotransferase (AST), alanine aminotransferase (ALT), and intestinal flora. A one - way analysis of variance was used for comparison of normally distributed continuous data between multiple groups, and the SNK - q test was used for further comparison between two groups. Results Compared with the CON group, the MOD group had a significant reduction in body weight and significant increases in AST and ALT (all P <0. 05), as well as large patchy necrosis of hepatocytes, significant increases in Verrucomicrobia, Akkermansia, and Erysipelatoclostridium, and significant reductions in Dubosiella and Duncaniella (all P <0. 05). Compared with the CON group, the ANFMT group had a significant increase in AST (P <0. 05), hepatocyte swelling and mild ballooning degeneration, significant increases in Unclassified and Faecalibaculum, and significant reductions in Patescibacteria, Deferribacteres, Muribaculum, Candidatus_Saccharimonas, Rikenella, Odoribacter, Mucispirillum, and Lachnospiraceae_unclassified (all P <0. 05). Compared with the MOD group, the FMT group had significant reductions in AST and ALT (both P < 0. 05), mild hepatocellular necrosis and marked ballooning degeneration, significant increases in Paramuribaculum and Bilophila, and significant reductions in Firmicutes, Rikenella, and Absiella (all P <0. 05). Conclusion Intestinal flora disturbance is observed in ACLF mice, and dysbacteriosis may lead to liver injury. FMT can alleviate liver inflammation in ACLF mice and thus exert a protective effect. [ABSTRACT FROM AUTHOR]

Objective To study the effects of different stimulation patterns on the pattern visual evoked potentials (PVEP) in mice with implanted electrodes onto the brain surface. Methods Twenty-two 6-week-old C57BL/6J mice were anesthetized and their heads were fixed on a stereotactic instrument. Recording electrodes were implanted onto the surface of the binocular area of the primary visual cortex, and the reference electrodes were implanted below the nasal bone. Three different forms of pattern visual stimulation were performed on each anaesthetized mouse, which were horizontal grating stimulation, vertical grating stimulation and reversal checkerboard stimulation. P100 waveform amplitudes and peak times of PVEP recorded under different pattern visual stimuli were compared. Results Under the same spatial frequency and temporal frequency, there were significant differences in the amplitudes (F=18.670, P＜0.01) and peak times (F=21.170， P＜0.01) of PVEP P100 waveforms evoked by horizontal grating stimulation (173.18 μV±33.00 μV, 87.59 ms±12.40 ms), vertical grating stimulation (189.05 μV±31.79 μV, 78.68 ms±9.50 ms) and reversal checkerboard stimulation (142.68 μV± 32.64 μV, 96.55 ms±17.57 ms). There were statistically significant differences in all the pairwise comparisons of the amplitudes and peak times of the PVEP P100 waveforms (P＜0.05). The coefficients of variation of the three groups of data were 19.06%, 16.82% and 22.87%, and 14.16%, 12.07% and 18.19%, respectively. Conclusion Vertical grating stimulation can be used to evoke pattern visual evoked potentials more sensitive and more stable than reversal checkerboard stimulation and horizontal grating stimulation. [ABSTRACT FROM AUTHOR]

Objective To explore the role of the age-related alteration of dopamine D1 receptor in nucleus accumbens (NAc) in the delayed emergence from isoflurane anesthesia in aged mice. Methods Firstly 8 aged mice (18-22 months) and 8 young mice (5-8 months) were extracted from 26 mice in each group. After exposure to 1.4% isoflurane, differences in the emergence time from isoflurane anesthesia and power spectrum of cortical electroencephalogram (EEG) were compared between the two groups. The rest of the mice in each group were used for microinjection. The effect of microinjecting dopamine D1 receptor agonist, antagonist and normal saline into the NAc on the emergence time and power spectrum were investigated during the recovery period after isoflurane anesthesia. Finally, the differences of expression levels of the NAc dopamine D1 receptor in aged and young mice were detected to analyze the potential mechanism underlying the delayed emergence after general anesthesia in aged mice. Results Compared with young mice, the emergence time from isoflurane anesthesia was notably prolonged in aged mice (P＜0.01). Power of δ band (＞1-4 Hz) in the cortical EEG was elevated in aged mice (P＜0.01), whilst the power of α band (＞8-12 Hz), β band (＞12-25 Hz) and γ band (＞25-60 Hz) were strikingly reduced (P＜0.05). Comparing with normal saline controls, microinjecting D1 receptor agonist significantly shortened the emergence time, reduced the power of δ band and enhanced the power of γ band in EEG of young mice (P＜ 0.01). Meanwhile, microinjecting D1 receptor antagonist prolonged the emergence time and increased the power in δ band in young mice (P＜0.01). However, microinjecting D1 receptor agonist/antagonist affected neither the emergence time nor cortical EEG in aged mice (P＞0.05). In addition, the expression level of D1 receptor in the NAc remarkably decreased in aged mice compared with that of young mice (P＜0.01). Conclusion The age-related downregulation of D1 receptor in the NAc attenuates its arousal-promoting ability for general anesthesia, which is probably a crucial reason for the delayed emergence in aged animals. [ABSTRACT FROM AUTHOR]

Objectives: We aimed to evaluate the biological effects of high static magnetic field (HiSMF, 2-12 Tesla [T]) exposure on mice in a stable and effective breeding environment in the chamber of a superconducting magnet.Methods: C57BL/6 mice were bred in the geomagnetic field and HiSMF with different magnetic field strengths (2-4 T, 6-8 T, and 10-12 T) for 28 days. The body weight, blood indices, organ coefficients, and histomorphology of major organs were analyzed.Results: The results showed that the HiSMF had no significant effect on the body weight, organ coefficients, or histomorphology of major organs in mice. The HiSMF had no effect on most routine blood and biochemical indices, but the value of the mean corpuscular hemoglobin (MCH) was increased in the 2-4 T group compared with that of the other groups, and the uric acid level (UA) was decreased in the three HiSMF groups compared with that of the control group.Conclusion: The C57BL/6 mice were not affected when they were exposed to different HiSMF environments for 28 days.Key Points: • No physiological problems were observed in mice with long-term whole-body exposure to HiSMF. [ABSTRACT FROM AUTHOR]

Obesity and other lifestyle diseases in modern society can be related to historical dietary changes from diets balanced in omega-6 and omega-3 to the unbalanced “Western-type” diet. It is recognized that diet influences the murine and human gut microbiome, and most research indicates that microbial diversity and composition are altered by high-fat diets (HFDs). However, good knowledge about the effects of early exposure to HFD on the maturation and structure of the bacterial community is limited. Using mice as model, we hypothesized that an HFD alters the early dynamic of the gut bacterial community toward an unstable/unhealthy state. By sequencing the V3 and V4 regions of the 16S ribosomal ribonucleic acid gene, we investigated the bacterial community in fecal samples of mice fed a control diet and an HFD at weaning (sampling time 1) and after 8 weeks of dietary intervention (11 weeks of age; sampling time 2). Natural temporal microbiome maturation was evidenced by a general increase in microbial diversity and shifts in microbial community between sampling times 1 and 2 toward a mature community. However, the HFD led to significant structural segregation of the microbiome compared with controls; the HFD diet repressed health-enhancing bacteria (eg, Bifidobacterium and Akkermansia ) and promoted health-detracting bacteria (ie, those associated with gut disorders, eg, Dorea ). We suggest that early-life consumption of HFD negatively impacts the natural gut bacterial community maturation leading toward a potentially persistent unhealthy stage. [ABSTRACT FROM AUTHOR]

Objective To investigate the effects of iridoid on the spermatogenesis in mice of type-2 diabetes induced by nicotinamide and streptozotocin. Methods Fifty-six male C57BL mice were randomly divided into seven groups (n=8 for each group): control group, diabetic group, diabetic and glibenclamide (0.25 mg/kg) group, diabetic and iridoid (20 and 40 mg/kg) group, and iridoid (20 and 40 mg/kg) group. Diabetic model was induced by intraperitoneal injection of streptozotocin (65 mg/kg) and nicotinamide (120 mg/kg). Glibenclamide and iridoid were orally fed for 28 days. Twenty-four hours after the last treatment, blood samples, testis and the epididymis were collected for detecting blood hormones, testicular biopsy, and the evaluation of sperm parameters. Results The body weights of diabetic mice were recovered from the reduction after treatment with 40 mg/kg of iridoid and glibenclamide（P＜0.05). The sperm count and viability were significantly decreased in diabetic group compared with those of control group (P＜0.05). The sperm count was significantly increased in diabetic and glibenclamide group than that of diabetic group (P＜0.05). The serum levels of lutein hormone (LH), follicle stimulating hormone (FSH) and testosterone (T) were significantly decreased in diabetic group compared with those of control group (P＜0.05). The serum levels of LH, FSH and T were increased after treatment with 40 mg/kg of iridoid and glibenclamide (P＜0.05). Conclusion These results suggest that iridoid can reduce the level of oxidative damage in diabetic mice, and effectively improve the fertility of mice. [ABSTRACT FROM AUTHOR]

Objective To explore the expression and significance of Rac1 and WAVE2 protein in glomerulus of high-fat diet induced C57BL/6J model mice. Methods Thirty-two male C57BL/6J mice (3-week old) were randomly assigned into two groups(16 in each group). The control group was fed with basic diet (10% fat) for 4 weeks. The high-fat diet group was fed with high-fat diet (60% fat) for 4 weeks. The kidney morphological changes were examined by HE and PAS staining. The expressions of Rac1 and WAVE2 protein were examined by Western blot and immunohistochemistry analysis. Results HE and PAS results showed that there were glomeruli mesangial matrix hyperplasia and exudation in high- fat diet group compared with control group. The immunohistochemistry and Western blotting results showed that expressions of Rac1 and WAVE2 in glomerulus were both increased in high-fat diet group compared with those of control group. Conclusion Rac1 and WAVE2 protein may be involved in glomerular injuries induced by high-fat diet. [ABSTRACT FROM AUTHOR]

Abstract: Aim: To gain insights into inter-strain differences in radiosensitivity. Methods: Mice of inbred strains, A/J, C57BL/6J, and C3H/HeMs, were irradiated at graded doses ranging from 20 to 60Gy. Skin reaction and leg contraction were observed for a period of 230 days and between 175 and 350 days, respectively. Gene expressions in leg skin tissue were quantified by quantitative RT-PCR assay at 1, 12 and 72h after 30Gy irradiation. Mice were locally irradiated by using a Cs-137 source. Results: The three strains showed various degrees of susceptibility to irradiation has evaluated by skin scores. Large inter-strain differences were also detected in the lengths of contraction. Expressions of several genes such as Per3 and Rad51ap1 displayed inter-strain differences. Conclusions: The continuum model of tissue injury revealed that genetic factor, which varies among strains, is one of the causes of variances in severity of damage after irradiation. [Copyright &y& Elsevier]

Objective To explore a simple and effective way of establishing a mice orthotopic tracheal transplantation model in studying bronchiolitis obliterans to reduce operation time. Methods Tracheas from C57BL/6 mice (n=30) were implanted into Balb/c mice (n=30). Grafts and hosts were matched according to body weight."Orotracheal intubation and endo-stent"was used for trachea anastomose and animal model was establish. Operating time was also noted. Grafts were harvested on Days 15, 30 and 60 after transplantation, using HE staining to observe pathological changes of allograft. Trachea blocking rate was also calculated. Results Until survival of trachea transplantation, the operative time of donor′s and the recipient′ tracheas resection were (7.0±1.0) min and (7.0±1.0) min respectively."Orotracheal intubation the endo-stent"establishment cost(1 ± 0.5)min. Trachea anastomose took(20.0 ± 4.0)min and the operation time last(43.0 ± 10.0)min. Conclusion The improvement of orthotopic tracheas transplantation using"Orotracheal intubation the endo-sten"for trachea anastomose is esay-operated and did required complicate skills and instruments with high successful rate. So it is an ideal and stable model in studying bronchiolitis obliterans. [ABSTRACT FROM AUTHOR]

Rat models of duodenogastric reflux have been used to study gastric stump cancer (GSC), but the underlying molecular mechanisms are poorly understood. Unlike rats, mice can be genetically modified, providing a superior model for studying the molecular mechanisms underlying GSC development, which is associated with duodenogastric reflux. This study aimed at developing a mouse model of duodenogastric reflux. C57BL/6 mice were randomly assigned to the control (n = 6), sham operation (n = 9), or gastrojejunostomy group (n = 12). Mice were sacrificed at 1, 3, and 6 months after surgery. Stomach tissue was stained with hematoxylin and eosin. Lesions were classified as chronic inflammation, intestinal metaplasia, or atypical hyperplasia. Nine mice underwent gastrojejunostomy without mortality. The animals in the gastrojejunostomy group exhibited chronic inflammation at 1, 3, and 6 months after surgery, showing intestinal metaplasia (n = 2) and atypical hyperplasia (n = 1) at 3 months and intestinal metaplasia (n = 2) and atypical hyperplasia (n = 2) at 6 months. The mice in the control group did not exhibit chronic inflammation or intestinal metaplasia, whereas those in the sham operation group exhibited chronic inflammation at 1, 3, and 6 months after surgery, without intestinal metaplasia or atypical hyperplasia. Intestinal metaplasia or atypical hyperplasia were more common in the gastrojejunostomy group than in the sham operation group (p = 0.012). A duodenogastric reflux mouse model can be created using gastrojejunostomy without gastrectomy. [ABSTRACT FROM AUTHOR]