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 liver 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 liver injury involve pathways related to xenobiotic metabolism and act via cytochrome P450,cytokine-cytokine receptor interaction,chemokine signaling,glutathione metabolism,and retinol metabolism.

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 liver 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 liver injury involve pathways related to xenobiotic metabolism and act via cytochrome P450,cytokine-cytokine receptor interaction,chemokine signaling,glutathione metabolism,and retinol metabolism.

Objective To investigate the effects and mechanisms of five-element music on the social behavior of the children of mothers with fear stress during pregnancy and provide a basis for the early prevention and treatment of clinical fetogenic affective disorders.Methods Forty-five pregnant mice were randomly divided into three groups:a control group,model group,and five-element music group.The model and five-element music group models were established using the bystander electric shock method.Additionally,the five-element music group was exposed to Palace Tune five-element music daily from 17:00 to 19:00 during pregnancy.On the 19th day of pregnancy,ELISA was employed to assess the levels of adrenocorticotropin(ACTH)and cortisol(CORT)in the serum of pregnant mice in each group for modeling evaluation.The offspring were subsequently grouped with their mother and underwent an 8-week-old three-box social experiment to observe their social behavior.We used the immunofluorescence double-labeling method to detect glutamatergic neuron activity in the medial prefrontal cortex(mPFC)of the offspring.High-performance liquid chromatography was employed to measure the total glutamate(Glu)content in the mPFC,while Gorky staining was used to observe changes in the dendritic spines of mPFC neurons in the offspring.Results Compared to those in the blank group,pregnant mice in the model group exhibited a significant increase in the levels of ACTH and CORT in their serum,and there was a significant decrease in the social interaction time and social novelty preference index of their offspring.There was also a significant decrease in glutamate neuron activity,glutamate content,and neuronal dendritic spine density.In contrast,compared with those in the model group,pregnant mice in the five-element music group demonstrated a reduction in the levels of ACTH and CORT in the serum,and there were improvements in the social behavior,glutamate neuron activity,glutamate content,and condition of neuronal dendritic spines in the offspring.Conclusions Intervention with five-element music effectively ameliorated the offspring's social behavior disorder result ing from prenatal fear stress;the mechanism was potentially linked to enhanced glutamate neuron activity in the mPFC region.

Objective: To examine the causal relationship between inflammatory factors and breast cancer using two-sample Mendelian randomization (MR) approach, so as to provide the basis for the prevention and treatment of breast cancer. Methods: Data of 91 inflammatory cytokines (n=14 824) and 5 subtypes of breast cancer (n=247 173) were collected from genome-wide association studies (GWAS). Single nucleotide polymorphism (SNP) associated with 91 inflammatory factors were selected as instrumental variables. MR analyses were performed using the inverse-variance weighted (IVW) method with inflammatory factors as exposure factors and breast cancer as outcome variables. The risk of type I error and the effect of multiple testing were reduced using the FDR correction method. The stability and reliability of the results were verified using Steiger test of directionality, MR-Egger regression, MR-PRESSO test and leave-one out method. Results: Twenty-three inflammatory factors, including β nerve growth factor, interleukin-5, cystatin D and C-X-C chemokine ligand 1 were statistically associated with breast cancer (all P<0.05). After FDR adjustment, only evaluated abundance of oncostatin-M was found to be statistically associated with an increased risk of Basal-like (triple-negative) breast cancer (OR=1.186, 95%CI: 1.081-1.302, P=0.001, q=0.029), and the other 22 inflammatory factors had a high risk of type I error (all q>0.1). The sensitivity analysis indicated that the results were robust. No instrumental variables were found to have a significant impact on the results, which could exclude the influence of heterogeneity, horizontal pleiotropy, and reverse causality on the outcome. Conclusion The increased abundance of oncostatin-M may increase the risk of Basal-like (triple-negative) breast cancer.

Ankylosing spondylitis is an autoimmune disease with a high incidence rate in clinic. It is characterized by abnormal ossification and ankylosis of the spine and sacroiliac joints. With the further development of the disease, the quality of life of patients will eventually decline sharply. At present, there are many treatment methods that can be chosen, and early intervention and comprehensive treatment are key, including drug therapy, rehabilitation training, surgical treatment, etc. Although there are many treatment methods to choose from, its pathogenesis is not fully understood, and it is still a difficult point in clinical treatment. This article systematically reviews the literature on the treatment of ankylosing spondylitis in recent years, providing reference for further optimizing comprehensive treatment strategies.

Objective:To explore the application and diagnostic value of low-dose scan technique of chest computed tomography(CT)combined with three dimensional(3D)reconstruction for ribs in chest trauma.Methods:A total of 118 patients with highly suspected rib fracture who admitted to the 904th Hospital of People's Liberation Army Joint Service Support Force were selected,and all cases underwent low-dose scan technique of chest CT combined with 3D reconstruction for ribs.The image qualities of chest CT scans with different low-doses combined with 3D reconstruction for ribs were analyzed,and the diagnostic accuracies among low dose scan technique of chest CT,3D reconstruction for ribs and the combination of them for chest trauma were compared.Results:Both the sharpness and clarity of the edges of the bronchi,blood vessels,lung parenchyma,interlobular septum,mediastinum and ribs were poorer,and the artifacts of soft tissue were more and the noise were more when the tube current of CT scan was 50 Ma.Both the sharpness and clarity of the edges of the bronchi,blood vessels,lung parenchyma,interlobular septum,mediastinum and ribs were general,and a part of soft tissues existed artifacts and the noise amounts were less when the tube current of CT scan was 70 Ma,which did not affect the diagnosis.The radiation dose as 50 mA was significantly higher than that as 70 mA,with a statistically significant difference(t=10.969,P<0.05).In 118 patients with chest trauma,the examination of low-dose scan technique of chest CT combined with 3D reconstruction for ribs indicated that there were 112 cases of rib fractures and 7 cases of costal cartilage fractures.In the examined 388 fractures of rib and costal cartilage,355 fractures(91.49%)were rib fractures and 33 fractures(8.51%)were costal cartilage fractures.In 118 patients with chest trauma,76 cases(64.41%)complicated with pulmonary contusions and lacerations,and 41 cases(35.75%)complicated with pleural effusion,and 10 cases(8.47%)complicated with thoracic vertebral fractures,and 6 cases(5.08%)complicated with splenic contusions and lacerations,and 5 cases(4.24%)complicated with mediastinal and subcutaneous emphysemas.The most direct imaging sign of rib fracture was visible and transparent low-density shadow.Chest CT scan can generally better display dislocation of the fractured end.The 3D reconstruction image showed a visibly line-like shadow on one side of rib if only one side of ribs fractured and the other side was intact.A total of 395 rib and costal cartilage fractures were confirmed by 3D reconstruction,which included 363 rib fractures(91.90%)and 32 costal cartilage fractures(8.10%).A total of 410 rib and costal cartilage fractures were confirmed by low-dose scan technique of chest CT combined with 3D reconstruction for ribs,which included 375 rib fractures(91.46%)and 35 costal cartilage fractures(8.54%).In the comparison of 418 rib injuries that were confirmed during surgery,the accuracy of low-dose scan technique of chest CT was 92.82%(388/418)in diagnosing rib and costal cartilage fractures,and the accuracy of 3D reconstruction for ribs was 94.50%(395/418)in diagnosing that,and the accuracy of low-dose scan technique of chest CT combined with 3D reconstruction for ribs was 95.69%(410/418)in diagnosing that.There was a significant difference in accuracy among the three types of examinations(x2=13.062,P<0.05).Conclusion:Low dose scan technique of chest CT combines with 3D reconstruction for ribs can be used in the diagnosis of chest trauma,which has higher accuracy and can provide reliable imaging information for clinical diagnosis and treatment.

Objective Transcriptome sequencing technology(RNA-seq)was used to analyze the mechanism of compound Dancao granules as an intervention for high-fat feed combined with carbon tetrachloride(CCl4)-induced non-alcoholic steatohepatitis.Methods 45 male C57BL/6J mice were split into two groups at random:normal control group,model control group,obeccholic acid group 10 mg/(kg·d),and compound Dancao granules low-and high-dose groups 3.74 g/(kg·d)and 7.48 g/(kg·d),with 9 mice in each group.Normal diet was made available to the control group,and the mice in the model group were given a high-fat diet combined with the subcutaneous injection of CC14,with 100％CC14 solution(4 mL/kg)in the first application,and 40％CC14-olive oil solution(2 mL/kg)in the second application,twice a week for a total of 6 weeks.Each drug group was administered the respective drug from week 3 for a total of 4 weeks.12 h after the last administration,the serum and liver tissues of mice in each group were collected,and a biochemical kit was used to detect serum liver function.Hematoxylin-eosin(HE),sirius scarlet,and oil red O staining were used to examine histopathological changes to the liver.The levels of IL-6,IL-10,TNF-α and TGF-β in mice liver were detected via ELISA,and the expression of α-SMA was observed by immunohistochemistry.Differential gene expression was analyzed by RNA-seq and functional enrichment analysis.To verify the differential expression of mRNA,quantitative reverse transcription PCR(qRT-PCR)was used.TDT-mediated dUTP nick-end labeling(TUNEL)staining was employed to identify apoptosis.Results The model control groups had significantly higher levels of serum alanine aminotransferase(ALT),aspartate aminotransferase(AST),total cholesterol(TC),and triglycerides(TG)than normal control group(P＜0.01).Additionally,there was obvious inflammatory cell infiltration in the liver tissue,collagen deposition in the sink and interlobule areas,and a significant increase in lipid droplet area(P＜0.01).The levels of IL-6 and TNF-α in liver tissue were significantly increased(P＜0.01),the levels of IL-10 and TGF-β were decreased(P＜0.01),and the expression of α-SMA was significantly increased(P＜0.01).The levels of TC,TG,ALT,and AST were significantly lower in groups that received compound Dancao granules and obeccholic acid than the model control group(P＜0.01),and inflammatory cell infiltration,collagen deposition,and fat accumulation in the sink and interlobule areas were improved(P＜0.01).The levels of IL-6 and TNF-α in liver tissue were significantly decreased(P＜0.01),the levels of IL-10 and TGF-β were increased(P＜0.05,P＜0.01),and the expression of α-SMA was significantly decreased(P＜0.01).RNA-seq sequencing result showed that 2819 genes in the normal control group were differentially expressed compared with the model control group,with 543 up-regulated and 2276 down-regulated genes.In a comparison of the model control group and compound Dancao granules group,240 genes were differentially expressed,including 206 up-regulated genes and 34 down-regulated genes.There were 221 genes with overlapping expression in the 2 groups and functional enrichment highlighted cell cycle(Cdt1,Plk1,Bub1b,Ttk,Knl1,Esco2,Cdc6,Ndc80,Cdc25b,Sgo1,Ccnb2,Espl1,Ccne1,Mcm4,Mcm5,Fbxo5,Bub1,Mcm2),apoptosis(Caspase3,Bax,P53,Apaf1,Bak,Caspase8),the P53 signaling pathway(P53,Ccnb2,Apaf1,Bak,Bax,Gtse1,Caspase3,Ccne1),arachidonic acid metabolism(Hpgds,Cyp2c54,Cyp2b10,Tbxas1,Cyp2c50),galactose metabolism(Hk3,Gla,Hk2,Akr1b7)and other signaling pathway genes.RNA-seq sequencing analysis showed that compound Danicao granules mainly regulated the apoptosis signaling pathway,and qRT-PCR confirmed that the mRNA expression of Caspase3,Bax,P53,Apaf1,Bak and Caspase8 in the liver tissue of the model control group was increased compared with that of the normal control group(P＜0.01).Compared with the model control group,the compound Dancao granules group showed decreased mRNA expression of Caspase3,Bax,P53,Apaf1,Bak and Caspase8 in liver tissue(P＜0.01).TUNEL staining showed that the number of cells showing nuclear shrinkage and apoptotic bodies decreased in the compound Dancao granule administration group.Conclusions Compound Dancao granules had a significant protective effect against non-alcoholic steatohepatitis induced by high-fat feed combined with CCl4,and its mechanism might be connected to the control of genes linked to apoptosis.

Objective Transcriptome sequencing technology(RNA-seq)was used to analyze the mechanism of compound Dancao granules as an intervention for high-fat feed combined with carbon tetrachloride(CCl4)-induced non-alcoholic steatohepatitis.Methods 45 male C57BL/6J mice were split into two groups at random:normal control group,model control group,obeccholic acid group 10 mg/(kg·d),and compound Dancao granules low-and high-dose groups 3.74 g/(kg·d)and 7.48 g/(kg·d),with 9 mice in each group.Normal diet was made available to the control group,and the mice in the model group were given a high-fat diet combined with the subcutaneous injection of CC14,with 100％CC14 solution(4 mL/kg)in the first application,and 40％CC14-olive oil solution(2 mL/kg)in the second application,twice a week for a total of 6 weeks.Each drug group was administered the respective drug from week 3 for a total of 4 weeks.12 h after the last administration,the serum and liver tissues of mice in each group were collected,and a biochemical kit was used to detect serum liver function.Hematoxylin-eosin(HE),sirius scarlet,and oil red O staining were used to examine histopathological changes to the liver.The levels of IL-6,IL-10,TNF-α and TGF-β in mice liver were detected via ELISA,and the expression of α-SMA was observed by immunohistochemistry.Differential gene expression was analyzed by RNA-seq and functional enrichment analysis.To verify the differential expression of mRNA,quantitative reverse transcription PCR(qRT-PCR)was used.TDT-mediated dUTP nick-end labeling(TUNEL)staining was employed to identify apoptosis.Results The model control groups had significantly higher levels of serum alanine aminotransferase(ALT),aspartate aminotransferase(AST),total cholesterol(TC),and triglycerides(TG)than normal control group(P＜0.01).Additionally,there was obvious inflammatory cell infiltration in the liver tissue,collagen deposition in the sink and interlobule areas,and a significant increase in lipid droplet area(P＜0.01).The levels of IL-6 and TNF-α in liver tissue were significantly increased(P＜0.01),the levels of IL-10 and TGF-β were decreased(P＜0.01),and the expression of α-SMA was significantly increased(P＜0.01).The levels of TC,TG,ALT,and AST were significantly lower in groups that received compound Dancao granules and obeccholic acid than the model control group(P＜0.01),and inflammatory cell infiltration,collagen deposition,and fat accumulation in the sink and interlobule areas were improved(P＜0.01).The levels of IL-6 and TNF-α in liver tissue were significantly decreased(P＜0.01),the levels of IL-10 and TGF-β were increased(P＜0.05,P＜0.01),and the expression of α-SMA was significantly decreased(P＜0.01).RNA-seq sequencing result showed that 2819 genes in the normal control group were differentially expressed compared with the model control group,with 543 up-regulated and 2276 down-regulated genes.In a comparison of the model control group and compound Dancao granules group,240 genes were differentially expressed,including 206 up-regulated genes and 34 down-regulated genes.There were 221 genes with overlapping expression in the 2 groups and functional enrichment highlighted cell cycle(Cdt1,Plk1,Bub1b,Ttk,Knl1,Esco2,Cdc6,Ndc80,Cdc25b,Sgo1,Ccnb2,Espl1,Ccne1,Mcm4,Mcm5,Fbxo5,Bub1,Mcm2),apoptosis(Caspase3,Bax,P53,Apaf1,Bak,Caspase8),the P53 signaling pathway(P53,Ccnb2,Apaf1,Bak,Bax,Gtse1,Caspase3,Ccne1),arachidonic acid metabolism(Hpgds,Cyp2c54,Cyp2b10,Tbxas1,Cyp2c50),galactose metabolism(Hk3,Gla,Hk2,Akr1b7)and other signaling pathway genes.RNA-seq sequencing analysis showed that compound Danicao granules mainly regulated the apoptosis signaling pathway,and qRT-PCR confirmed that the mRNA expression of Caspase3,Bax,P53,Apaf1,Bak and Caspase8 in the liver tissue of the model control group was increased compared with that of the normal control group(P＜0.01).Compared with the model control group,the compound Dancao granules group showed decreased mRNA expression of Caspase3,Bax,P53,Apaf1,Bak and Caspase8 in liver tissue(P＜0.01).TUNEL staining showed that the number of cells showing nuclear shrinkage and apoptotic bodies decreased in the compound Dancao granule administration group.Conclusions Compound Dancao granules had a significant protective effect against non-alcoholic steatohepatitis induced by high-fat feed combined with CCl4,and its mechanism might be connected to the control of genes linked to apoptosis.

Alcoholic liver disease (ALD) is a growing global health concern, and its early pathogenesis includes steatosis and steatohepatitis. Inhibiting lipid accumulation and inflammation is a crucial step in relieving ALD. Evidence shows that puerarin (Pue), an isoflavone isolated from Pueraria lobata, exerts cardio-protective, neuroprotective, anti-inflammatory, antioxidant activities. However, the therapeutic potential of Pue on ALD remains unknown. In the study, both the NIAAA model and ethanol (EtOH)-induced AML-12 cell were used to explore the protective effect of Pue on alcoholic liver injury in vivo and in vitro and related mechanism. The results showed that Pue (100 mg·kg^-1) attenuated EtOH-induced liver injury and inhibited the levels of SREBP-1c, TNF-α, IL-6 and IL-1β, compared with silymarin (Sil, 100 mg·kg^-1). In vitro results were consistent within vivo results. Mechanistically, Pue might suppress liver lipid accumulation and inflammation by regulating MMP8. In conclusion, Pue might be a promising clinical candidate for ALD treatment.

Background::Sepsis is a systemic inflammatory syndrome induced by several infectious agents. Multiple organs are affected by sepsis, including the liver, which plays an important role in metabolism and immune homeostasis. Fibroblast growth factors (FGFs) participate in several biological processes, although the role of FGF5 in sepsis is unclear. Methods::In this study, lipopolysaccharide (LPS) was administrated to mice to establish a sepsis-induced liver injury. A similar in vitro study was conducted using L-02 hepatocytes. Western blot and immunohistochemistry staining were performed to evaluate the FGF5 expression level in liver tissues and cells. Inflammatory cell infiltrations, cleaved-caspase-3 expressions, reactive oxygen species and levels of inflammatory cytokines were detected by immunofluorescence, dihydroethidium staining, and reverse transcription quantitative polymerase chain reaction analysis, respectively. Flow cytometry was used to detect the apoptosis level of cells. In addition, ribonucleic acid (RNA)-sequencing was applied to explore the possible mechanism by which FGF5 exerted effects. Results::LPS administration caused FGF5 down-regulation in the mouse liver as well as in L-02 hepatocytes. Additionally, with FGF5 overexpression, liver injury and the level of hepatocyte apoptosis were ameliorated. Further, RNA sequencing performed in hepatocytes revealed the phosphoinositide-3-kinase/protein kinase B (PI3K/AKT) pathway as a possible pathway regulated by FGF5. This was supported using an inhibitor of the PI3K/AKT pathway, which abrogated the protective effect of FGF5 in LPS-induced hepatocyte injury. Conclusion::The anti-apoptotic effect of FGF5 on hepatocytes suffering from LPS has been demonstrated and was dependent on the activation of the PI3K/AKT signaling pathway.