Based on network pharmacology,molecular docking,and experimental validation,this study explored the mechanisms of Geranium wilfordii Maxim(GWM)in the treatment of liver injury.Mice were randomly divided into a control group(CON group),a model group(CCl4 group),a high-dose drug group(GWM-H group),and a low-dose group(GWM-L group).The liv-er injury model in mice was induced by CCl4,and liver tissue pathological morphology was ob-served,along with the measurement of the relative gene expression levels of liver inflammatory factors.Active ingredients of traditional Chinese medicine and target information of Chinese medi-cine and diseases were obtained through databases such as TCMSP,PubChem,Swiss Target Pre-diction,Super-PRED,Gene Cards,and DisGeNET.Intersecting the targets of liver injury,necropto-sis,and drugs yielded potential drug targets.String database was used for protein-protein interac-tion(PPI)analysis of the potential targets.Furthermore,Cytoscape was utilized to construct a net-work diagram of"drug-disease-active ingredient-intersection target,"Wei Sheng Xin was used for GO and KEGG pathway analysis.Molecular docking was performed using MOE software,and the results of molecular docking were experimentally validated to detect the expression of key targets in the RIPK1/RIPK3/MLKL signaling pathway.Animal experiments showed that compared to the CON group,the CCl4 group of mice exhibited a significant increase in liver organ index(P＜0.05),markedly elevated serum AST activity(P＜0.05),and a highly significant increase in ALT activity(P＜0.01).Pathological examination revealed chaotic liver lobules,severe hepatocyte steatosis,ex-tensive hepatocyte necrosis,and inflammatory cell infiltration in the livers of mice in the CCl4 group.In comparison to the CCl4 group,the GWM-H group showed a significant decrease in liver organ index(P＜0.05),while the GWM-L group displayed a downward trend.The GWM-H group exhibited a significant reduction in serum AST activity(P＜0.05),the GWM-L group showed a decreasing trend in serum AST activity,the GWM-H group demonstrated a highly significant de-crease in serum ALT activity(P＜0.01),and the GWM-L group displayed a significant decrease in serum ALT activity(P＜0.05).Histopathological examination revealed that the drug treatment groups could improve CCl4-induced liver injury,with the GWM-H group showing better efficacy than the GWM-L group.RT-qPCR results of liver tissues showed that compared to the CON group,the CCl4 group exhibited a highly significant increase in the relative expression of IL-1βand PGE2 mRNA(P＜0.01),while the mRNA relative expression of COX2 showed an increasing trend.In contrast,compared to the CCl4 group,the GWM-H group showed a remarkably significant decrease in the relative expression of IL-1βmRNA(P＜0.01),a significant decrease in PGE2 mR-NA expression(P＜0.05),and a decreasing trend in COX2 mRNA expression.Through network pharmacology,56 potential targets related to GWM in ameliorating necroptosis-induced liver injury were identified.Key targets,based on degree value,include TNF,Bcl2,HSP90AA1,and Caspase8,while the key components are quercetin,luteolin,kaempferol,and ellagic acid.Functional enrich-ment analysis yielded 2 173 entries for GO and 146 biological pathways for KEGG.Molecular doc-king results indicated a strong binding capacity between the main components of GWM and key targets.RT-qPCR experimental results showed that compared to the CON group,the CCl4 group exhibited a extremely significantly increase in the mRNA relative expression of TNF-α,TNFR1,MLKL(P＜0.01),significantly increase in the mRNA relative expression of FAS,RIPK1,RIPK3 mRNA(P＜0.05),and a significant decrease in Caspase 8 mRNA expression(P＜0.05).The addi-tion of GWM successfully reversed this trend;compared to the CCl4 group,the GWM-H group showed a highly significant decrease in the mRNA relative expression of TNF-α,TNFR1,FAS and MLKL mRNA(P＜0.01),significant decrease in RIPK1,RIPK3 mRNA expression(P＜0.05),and an increasing trend in CASPASE8 mRNA expression.GWM exerts hepatoprotective effects through multiple components and pathways,among which inhibition of the RIPK1/RIPK3/MLKL pathway reduces hepatocyte necroptosis,potentially serving as one of the essential mechanisms for its protective effects.

Based on network pharmacology,molecular docking,and experimental validation,this study explored the mechanisms of Geranium wilfordii Maxim(GWM)in the treatment of liver injury.Mice were randomly divided into a control group(CON group),a model group(CCl4 group),a high-dose drug group(GWM-H group),and a low-dose group(GWM-L group).The liv-er injury model in mice was induced by CCl4,and liver tissue pathological morphology was ob-served,along with the measurement of the relative gene expression levels of liver inflammatory factors.Active ingredients of traditional Chinese medicine and target information of Chinese medi-cine and diseases were obtained through databases such as TCMSP,PubChem,Swiss Target Pre-diction,Super-PRED,Gene Cards,and DisGeNET.Intersecting the targets of liver injury,necropto-sis,and drugs yielded potential drug targets.String database was used for protein-protein interac-tion(PPI)analysis of the potential targets.Furthermore,Cytoscape was utilized to construct a net-work diagram of"drug-disease-active ingredient-intersection target,"Wei Sheng Xin was used for GO and KEGG pathway analysis.Molecular docking was performed using MOE software,and the results of molecular docking were experimentally validated to detect the expression of key targets in the RIPK1/RIPK3/MLKL signaling pathway.Animal experiments showed that compared to the CON group,the CCl4 group of mice exhibited a significant increase in liver organ index(P＜0.05),markedly elevated serum AST activity(P＜0.05),and a highly significant increase in ALT activity(P＜0.01).Pathological examination revealed chaotic liver lobules,severe hepatocyte steatosis,ex-tensive hepatocyte necrosis,and inflammatory cell infiltration in the livers of mice in the CCl4 group.In comparison to the CCl4 group,the GWM-H group showed a significant decrease in liver organ index(P＜0.05),while the GWM-L group displayed a downward trend.The GWM-H group exhibited a significant reduction in serum AST activity(P＜0.05),the GWM-L group showed a decreasing trend in serum AST activity,the GWM-H group demonstrated a highly significant de-crease in serum ALT activity(P＜0.01),and the GWM-L group displayed a significant decrease in serum ALT activity(P＜0.05).Histopathological examination revealed that the drug treatment groups could improve CCl4-induced liver injury,with the GWM-H group showing better efficacy than the GWM-L group.RT-qPCR results of liver tissues showed that compared to the CON group,the CCl4 group exhibited a highly significant increase in the relative expression of IL-1βand PGE2 mRNA(P＜0.01),while the mRNA relative expression of COX2 showed an increasing trend.In contrast,compared to the CCl4 group,the GWM-H group showed a remarkably significant decrease in the relative expression of IL-1βmRNA(P＜0.01),a significant decrease in PGE2 mR-NA expression(P＜0.05),and a decreasing trend in COX2 mRNA expression.Through network pharmacology,56 potential targets related to GWM in ameliorating necroptosis-induced liver injury were identified.Key targets,based on degree value,include TNF,Bcl2,HSP90AA1,and Caspase8,while the key components are quercetin,luteolin,kaempferol,and ellagic acid.Functional enrich-ment analysis yielded 2 173 entries for GO and 146 biological pathways for KEGG.Molecular doc-king results indicated a strong binding capacity between the main components of GWM and key targets.RT-qPCR experimental results showed that compared to the CON group,the CCl4 group exhibited a extremely significantly increase in the mRNA relative expression of TNF-α,TNFR1,MLKL(P＜0.01),significantly increase in the mRNA relative expression of FAS,RIPK1,RIPK3 mRNA(P＜0.05),and a significant decrease in Caspase 8 mRNA expression(P＜0.05).The addi-tion of GWM successfully reversed this trend;compared to the CCl4 group,the GWM-H group showed a highly significant decrease in the mRNA relative expression of TNF-α,TNFR1,FAS and MLKL mRNA(P＜0.01),significant decrease in RIPK1,RIPK3 mRNA expression(P＜0.05),and an increasing trend in CASPASE8 mRNA expression.GWM exerts hepatoprotective effects through multiple components and pathways,among which inhibition of the RIPK1/RIPK3/MLKL pathway reduces hepatocyte necroptosis,potentially serving as one of the essential mechanisms for its protective effects.

OBJECTIVE:Researches show that a combination of platelet-rich plasma and adipose-derived stem cells can accelerate the healing of skin lesions.However,systematic evidence for the combination of the two is still lacking.The purpose of this study was to assess the efficacy of a combination of two interventions in a clinical rodent skin wound model.METHODS:We searched PubMed,Embase,Cochrane,and CNKI and selected the studies of platelet-rich plasma,adipose-derived stem cell transplantation,or their combination on skin wounds in experimental animals published until July 2023.Wound healing and wound transformation growth factor β,CD31,type Ⅰ collagen,and vascular endothelial growth factor were used as indicators.RevMan 5.3 and Stata 15.0 were used to analyze the data.RESULTS:A total of 12 studies were included,of which 8 studies used rats as experimental subjects and 4 studies used mice as experimental subjects.The experimental group was treated with platelet-rich plasma combined with adipose-stem cell transplantation,and the control group was treated with platelet-rich plasma alone.The results of meta-analysis showed that the wound healing rate of the experimental group at 3,7,and 10 days after treatment was greater than that of the control group[SMD=2.65,95％CI(1.29,4.01),Z=3.81,P=0.0001;SMD=3.38,95％CI(2.47,4.30),Z=7.24,P<0.00001;SMD=2.62,95％CI(1.50,3.73),Z=4.61,P<0.00001].The wound healing time of the experimental group was shorter than that of the control group[SMD=-2.12,95％CI(-3.5,-0.74),P=0.003].The expression of transforming growth factor β,positive rate of CD31,expression of type Ⅰ collagen,and vascular endothelial growth factor in wound of experimental group were higher than those of control group[SMD=5.65,95％CI(1.22,10.08),Z=2.50,P=0.01;SMD=2.49,95％CI(1.96,3.02),Z=9.28,P<0.00001;SMD=3.44,95％CI(0.72,6.17),Z=2.48,P=0.01;SMD=2.38,95％CI(0.97,3.79),Z=3.30,P=0.0010].CONCLUSION:Our results show that platelet-rich plasma+adipose-derived stem cells combined treatment can improve the wound healing rate,shorten the wound healing time,and at the same time increase the expression of transforming growth factor β,CD31,type Ⅰ collagen,and vascular endothelial growth factor to accelerate healing.Due to the limitations of the model,more animal testing and clinical trials are needed.

Objective:This study aimed to compare short-chain fatty acid (SCFA) levels in saliva between patients with laryngopharyngeal reflux disease (LPRD) and healthy controls, and to explore the relationship between these SCFAs and the salivary microbiota.Methods:A retrospective case-control study was conducted, enrolling 36 patients with laryngopharyngeal reflux disease (LPRD) who visited the Department of Otorhinolaryngology Head and Neck Surgery, the Eighth Medical Center, Chinese PLA General Hospital between February and November 2023. All patients were diagnosed via pharyngeal pH monitoring. The LPRD group included 30 males and 6 females, aged 20-53 years (30.61±7.83 years). In addition, 39 healthy volunteers were recruited as the control group, comprising 25 males and 14 females, aged 18–58 years (28.64±7.97 years). Unstimulated mixed saliva samples were collected from all participants. Concentrations of eight SCFAs (acetic acid, propionic acid, isobutyric acid, butyric acid, valeric acid, isovaleric acid, hexanoic acid, and heptanoic acid) in saliva were quantified using gas chromatography-mass spectrometry (GC-MS). Salivary DNA was extracted, followed by amplification and sequencing of the 16S rRNA gene to analyze the microbiota composition at the genus level. The SCFA concentrations and the differences in bacterial species between the LPRD and control groups were compared, and the correlation between SCFA concentrations and the relative abundance of different bacterial genera in the salivary microbiota was analyzed. All statistical analyses were performed using R version 3.6.1 and SPSS version 26.0, while, microbiome analyses were conducted using R language.Results:Salivary hexanoic acid concentration was significantly higher in the LPRD group than in the control group [(29.50±19.61) ng/ml vs. (10.15±3.65) ng/ml; t=-2.72, P<0.05]. Significant differences in the relative abundance of 17 bacterial genera were observed between the two groups ( P<0.05), including Prevotella, Butyrivibrio, Streptococcus, and Actinomyces. Correlation analysis revealed that hexanoic acid concentration was significantly positively correlated with the abundance of Butyrivibrio (γ=0.73, P<0.05) and Streptococcus (γ=0.78, P<0.05), while showing a significant negative correlation with Actinomyces (γ=-0.73, P<0.05). Conclusion:Elevated salivary hexanoic acid levels may be associated with the development of LPRD. Dysbiosis of the salivary microbiota might contribute to LPRD pathogenesis by altering the concentrations of SCFA, particularly hexanoic acid.

OBJECTIVE:Researches show that a combination of platelet-rich plasma and adipose-derived stem cells can accelerate the healing of skin lesions.However,systematic evidence for the combination of the two is still lacking.The purpose of this study was to assess the efficacy of a combination of two interventions in a clinical rodent skin wound model.METHODS:We searched PubMed,Embase,Cochrane,and CNKI and selected the studies of platelet-rich plasma,adipose-derived stem cell transplantation,or their combination on skin wounds in experimental animals published until July 2023.Wound healing and wound transformation growth factor β,CD31,type Ⅰ collagen,and vascular endothelial growth factor were used as indicators.RevMan 5.3 and Stata 15.0 were used to analyze the data.RESULTS:A total of 12 studies were included,of which 8 studies used rats as experimental subjects and 4 studies used mice as experimental subjects.The experimental group was treated with platelet-rich plasma combined with adipose-stem cell transplantation,and the control group was treated with platelet-rich plasma alone.The results of meta-analysis showed that the wound healing rate of the experimental group at 3,7,and 10 days after treatment was greater than that of the control group[SMD=2.65,95％CI(1.29,4.01),Z=3.81,P=0.0001;SMD=3.38,95％CI(2.47,4.30),Z=7.24,P<0.00001;SMD=2.62,95％CI(1.50,3.73),Z=4.61,P<0.00001].The wound healing time of the experimental group was shorter than that of the control group[SMD=-2.12,95％CI(-3.5,-0.74),P=0.003].The expression of transforming growth factor β,positive rate of CD31,expression of type Ⅰ collagen,and vascular endothelial growth factor in wound of experimental group were higher than those of control group[SMD=5.65,95％CI(1.22,10.08),Z=2.50,P=0.01;SMD=2.49,95％CI(1.96,3.02),Z=9.28,P<0.00001;SMD=3.44,95％CI(0.72,6.17),Z=2.48,P=0.01;SMD=2.38,95％CI(0.97,3.79),Z=3.30,P=0.0010].CONCLUSION:Our results show that platelet-rich plasma+adipose-derived stem cells combined treatment can improve the wound healing rate,shorten the wound healing time,and at the same time increase the expression of transforming growth factor β,CD31,type Ⅰ collagen,and vascular endothelial growth factor to accelerate healing.Due to the limitations of the model,more animal testing and clinical trials are needed.

Objective:This study aimed to compare short-chain fatty acid (SCFA) levels in saliva between patients with laryngopharyngeal reflux disease (LPRD) and healthy controls, and to explore the relationship between these SCFAs and the salivary microbiota.Methods:A retrospective case-control study was conducted, enrolling 36 patients with laryngopharyngeal reflux disease (LPRD) who visited the Department of Otorhinolaryngology Head and Neck Surgery, the Eighth Medical Center, Chinese PLA General Hospital between February and November 2023. All patients were diagnosed via pharyngeal pH monitoring. The LPRD group included 30 males and 6 females, aged 20-53 years (30.61±7.83 years). In addition, 39 healthy volunteers were recruited as the control group, comprising 25 males and 14 females, aged 18–58 years (28.64±7.97 years). Unstimulated mixed saliva samples were collected from all participants. Concentrations of eight SCFAs (acetic acid, propionic acid, isobutyric acid, butyric acid, valeric acid, isovaleric acid, hexanoic acid, and heptanoic acid) in saliva were quantified using gas chromatography-mass spectrometry (GC-MS). Salivary DNA was extracted, followed by amplification and sequencing of the 16S rRNA gene to analyze the microbiota composition at the genus level. The SCFA concentrations and the differences in bacterial species between the LPRD and control groups were compared, and the correlation between SCFA concentrations and the relative abundance of different bacterial genera in the salivary microbiota was analyzed. All statistical analyses were performed using R version 3.6.1 and SPSS version 26.0, while, microbiome analyses were conducted using R language.Results:Salivary hexanoic acid concentration was significantly higher in the LPRD group than in the control group [(29.50±19.61) ng/ml vs. (10.15±3.65) ng/ml; t=-2.72, P<0.05]. Significant differences in the relative abundance of 17 bacterial genera were observed between the two groups ( P<0.05), including Prevotella, Butyrivibrio, Streptococcus, and Actinomyces. Correlation analysis revealed that hexanoic acid concentration was significantly positively correlated with the abundance of Butyrivibrio (γ=0.73, P<0.05) and Streptococcus (γ=0.78, P<0.05), while showing a significant negative correlation with Actinomyces (γ=-0.73, P<0.05). Conclusion:Elevated salivary hexanoic acid levels may be associated with the development of LPRD. Dysbiosis of the salivary microbiota might contribute to LPRD pathogenesis by altering the concentrations of SCFA, particularly hexanoic acid.

Objective To investigate the potential significance of FOXP3 expression in BRCA1/2-mutant breast cancer.Methods A total of 48 BRCA mutation carriers(16 with BRCA1 and 32 with BRCA2)and 78 age-matched non-carriers were included in this study.Immunohistochemistry was used to detect the expression of FOXP3 in breast cancer tissues.The FOXP3 RNA expression in 39 BRCA1,36 BRCA2,and 948 non-carrier breast cancer patients from TCGA-BRCA and the correlation with homologous recombin-ation deficiency scores were evaluated to validate the immunohistochemistry results.Results The FOXP3 positive rate was 43.8%(7/16)in BRCA1 mutation carriers,59.4%(19/32)in BRCA2 mutation carriers,and 9.0%(7/78)in non-carriers.The FOXP3 positive rates in patients with BRCA1/2 mutant breast cancer were significantly higher than those in non-carriers(P=0.002;P<0.001).TCGA-BRCA results showed that the FOXP3 RNA level in BRCA1/2 mutant breast cancer was significantly higher than that in non-carriers(P=0.02,P=0.004).The FOXP3 RNA level was positively correlated with the homologous recombination deficiency score(Spearman R=0.30,P<2.2e-16).Conclusion Patients with BRCA1/2 mutant breast cancers have higher FOXP3expression than non-carriers,and may be more sensitive to immunotherapy.

Objective To invesigate the feasibility of low radiation dose and low contrast dosage in triple-rule-out computed tomo-graphy angiography(TRO-CTA)on the 320-row detector CT.Methods A total of 120 patients who underwent CTA were prospec-tively selected.All patients were divided into control group(n=90)and experimental group(n=30).The control group employed standard-doses protocol of pulmonary CTA(120 kV tube voltage,45 mL contrast dosage),coronary CTA(120 kV,50-60 mL),and aortic CTA(120 kV,75 mL),while the experimental group received TRO-CTA with 100 kV and 70-80 mL.The peak time of contrast dosage at the pulmonary artery and aorta was measured by low-dose detection method in the experimental group,and the contrast examination was performed sequentially in the control group.Subjective scores and objective image quality of the pulmonary artery,coronary artery,and aorta in the experimental group and the control group were measured and compared,respectively.The effective dose(ED)between the two groups were recorded and compared.Independent samples t-test and Fisher exact probability were used to analyze the statistical differences between the above measures.Results There were no significant differences in CT values,con-trast-to-noise ratio(CNR),signal-to-noise ratio(SNR)of pulmonary artery,coronary artery and aorta between the two groups(P＞0.05).The mean subjective scores of pulmonary artery,coronary artery and aorta segments in the two groups were not less than 3 points,meeting the requirement of clinical diagnosis.There was no statistical difference in subjective scores between the two groups(P＞0.05).There was statistically significant difference in ED between the two groups(P＜0.05).The ED of pulmonary artery,coronary artery,and aorta in the experimental group were 11.49％,13.33％,and 11.46％significantly lower than those in the control group,respec-tively.Conclusion It is feasible to obtain TRO-CTA images used by the low radiation dose and low contrast dosage on the 320-row detector CT,and radiation dose and contrast dosage can be reduced reasonably without alterations of TRO-CTA images quality in clinical practice.

Objective:To investigate the prognosis and the factors that influence it in patients with non-gastric gastrointestinal stromal tumors (GISTs) who are at low risk of recurrence.Methods:This was a retrospective cohort study. Clinicopathologic and prognostic data from patients with non-gastric GISTs and at low risk of recurrence (i.e., very low-risk or low-risk according to the 2008 version of the Modified NIH Risk Classification), who attended 18 medical centers in China between January 2000 and June 2023, were collected. We excluded patients with a history of prior malignancy, concurrent primary malignancy, multiple GISTs, and those who had received preoperative imatinib. The study cohort comprised 1,571 patients with GISTs, 370 (23.6%) of whom were at very low-risk and 1,201 (76.4%) at low-risk of recurrence. The cohort included 799 (50.9%) men and 772 (49.1%) women of median age 57 (16–93) years. Patients were followed up to July 2024. The prognosis and its influencing factors were analyzed. Receiver operating characteristic curves for tumor diameter and Ki67 were established, and the sensitivity, specificity, area under the curve (AUC) and optimal cut-off value with 95% confidence intervals were calculated. Propensity score matching was implemented using the 1:1 nearest neighbor matching method with a matching tolerance of 0.02.Results:With a median follow-up of 63 (12–267) months, the 5- and 10-year overall survival (OS) rates of the 1,571 patients were 99.5% and 98.0%, respectively, and the 5- and 10-year disease-free survival (DFS) rates were 96.3% and 94.4%, respectively. During postoperative follow-up, 3.8% (60/1,571) patients had disease recurrence or metastasis, comprising 0.8% (3/370) in the very low-risk group and 4.7% (57/1,201) in the low-risk group. In the low-risk group, recurrence or metastasis occurred in 5.5% (25/457) of patients with duodenal GISTs, 3.9% (25/645) of those with small intestinal GISTs, 9.2% (6/65) of those with rectal GISTs, and 10.0% (1/10) of those with colonic GISTs. Among the 60 patients with metastases, 56.7% (34/60) of the metastases were located in the abdominal cavity, 53.3% (32/60) in the liver, and 3.3% (2/60) in bone. During the follow-up period, 13 patients (0.8%) died of disease. Receiver operating characteristic curves were plotted for tumor diameter and Ki67 and assessed using the Jordon index. This showed that the difference in DFS between the two groups was statistically significant when the cutoff value for tumor diameter was 3.5 cm (AUC 0.731, 95% CI: 0.670–0.793, sensitivity 77.7%, specificity 64.1%). Furthermore, the difference in DFS between the two groups was statistically significant when the cutoff value for Ki67 was 5% (AUC 0.693, 95% CI: 0.624–0.762, sensitivity 60.7%, specificity 65.3%). Multifactorial analysis revealed that tumor diameter ≥3.5 cm, Ki67 ≥5%, and R1 resection were independent risk factors for DFS in patients with non-gastric GISTs at low risk of recurrence (all P<0.05). Furthermore, age >57 years, Ki67 ≥5%, and R1 resection were also independent risk factors for OS in patients with non-gastric GISTs at low risk of recurrence (all P<0.05). We also grouped the patients according to whether they had received postoperative adjuvant treatment with imatinib for 1 or 3 years. This yielded 137 patients in the less than 1-year group, 139 in the 1-year plus group; and 44 in both the less than 3 years and 3-years plus group. After propensity score matching for age, tumor diameter, Ki67, and resection status, the differences in survival between the two groups were not statistically significant (all P>0.05). The 10-year DFS and OS were 87.5% and 95.5%, respectively, in the group treated with imatinib for less than 1 year and 88.5% and 97.8%, respectively, in the group treated for more than 1 year. The 10-year DFS and OS were 89.6% and 92.6%, respectively, in the group treated with imatinib for less than 3 years and 88.0% and 100.0%, respectively, in the group treated with imatinib for more than 3 years. Conclusion:The overall prognosis of primary, non-gastric, low recurrence risk GISTs is relatively favorable; however, recurrences and metastases do occur. Age, tumor diameter, Ki67, and R1 resection may affect the prognosis. For some patients with low risk GISTs, administration of adjuvant therapy with imatinib for an appropriate duration may help prevent recurrence and improve survival.

Objective To explore the effects of long non-coding RNA(lncRNA)HEM2M overexpression on liver injury in mice with non-alcoholic fatty liver disease(NAFLD).Methods Wild-type C57BL/6(WT)mice and myeloid cell-specific HEM2M knock-in(MYKI)mice were fed normal(ND)or high-fat diet(HFD)for 12 weeks.After intraperitoneal glucose tolerance and insulin tolerance tests,the mice were euthanized for detection of liver function indicators in the serum and liver tissue.HE staining and F4/80 immunohistochemical staining were used to examine liver pathologies,and the levels of IL-6,IL-1β,and TNF-α in the liver tissues were determined with ELISA.The mRNA expressions of HEM2M and the markers of M1 macrophages(TNF-α,iNOS,and IL-6)and M2 macrophages(Arg-1,YM-1,and IL-10)were detected using qRT-PCR,and the protein expressions of P-AKT,T-AKT,NLRC4,caspase-1 and GSDMD were assayed using immunoblotting.Caspase-1 activity in the liver tissues was determined with colorimetric measurement and immunofluorescence assay.Results Compared with HFD-fed WT mice,MYKI mice with HFD feeding showed milder liver function damage(P<0.01),alleviated hepatic steatosis,and reduced liver macrophage infiltration,glucose tolerance impairment and insulin resistance(P<0.01).The levels of IL-6,IL-1β,and TNF-α and mRNA expressions of M1 type macrophage markers were significantly decreased(P<0.01)and those of M2 type markers increased(P<0.01)in the liver tissues of HFD-fed MYKI mice,which also showed reduced NLRC4 inflammasome activity,caspase-1 activation,and GSDMD-N protein expression compared with their WT counterparts(P<0.05).Conclusion Overexpression of HEM2M reduces the production of hepatic inflammatory factors,improves insulin resistance and inhibits hepatic NLRC4 inflammasome activation,which leads to reduced hepatic pyroptosis and liver injury in NAFLD mice.