1.Clinical features and lymphocyte subtypes in patients with IgG 4-related diseases
Liping GUO ; Weirong WANG ; Jiangpeng LIU ; Bangmao WANG ; Lu ZHOU
Chinese Journal of Internal Medicine 2024;63(4):394-400
Objective:To deepen understanding of IgG 4-related diseases (RDs), we analyzed the associated lymphocyte subtypes, and explored the pathogenesis and potential immunotherapeutic targets. Methods:Eighty-six patients with IgG 4-RDs were enrolled, and their clinical characteristics, peripheral lymphocyte subtypes, and disease course were analyzed. Results:The mean age of the participants was 36-87(62±11) years; 51 were male (59.3%) and 35 were women (40.7%); and 34.9% had a history of allergy. Follow-up lasted 4.8 (0.4, 14.1) months. The most common symptoms were abdominal pain, and submandibular gland and lacrimal gland swelling (each 20.9%). Sixty-five (75.6%) participants had multiple organ involvement, and the most frequently affected organs were the pancreas (52.3%), submandibular gland (51.2%), and lacrimal gland (34.9%). A high eosinophil count; high IgE, IgG, IgG 1, and IgG 4 concentrations; and low complement C3 and C4 concentrations were present in 18.8% (16/85), 30.0% (24/80), 72.9% (62/85), 58.3% (28/48), 89.5% (77/86), 61.2% (52/85), and 50.0% (42/84), respectively, of the participants. In addition, 64.7% (55/85) were positive for autoantibodies, and the most frequent was anti-nuclear antibody (63.5%). The proportion of CD4 +T lymphocytes increased in 25.7% (9/35) of the participants, which was accompanied by an increase in the ratio of CD4 +/CD8 +T lymphocytes (22.9%, 8/35). Importantly, most participants (90.0%, 18/20) had a high proportion of regulatory T (Treg) cells. High interleukin (IL)-2, IL-6, and IL-10 concentrations were present in 50.0% (11/22), 33.3% (10/30), and 16.7% (5/30), respectively, of the participants. Substantial lymphoplasmacytic infiltration, fibrosis, IgG 4-positive plasma cell infiltration, and lymphoid follicle hyperplasia or ectopic formation were present in 79.2% (42/53), 67.9%(36/53), 35.8%(19/53) and 30.2% (16/53), respectively, of the participants. Fifty-three participants with detailed pathologic data were also further evaluated, of whom 24.5% (13/53), 3.8% (2/53), and 67.9% (36/53) had definite, probable, and possible diagnoses; and 3.8% (2/53) could not be diagnosed. Compared with baseline, the percentage of eosinophils and the IgE, IgG, and IgG 4 concentrations decreased significantly; and the complement C3 and C4 concentrations had increased significantly after 6 months of treatment (all P<0.05). The IgG 4 concentration after 6 months of treatment negatively correlated with that of C4, and positively correlated with the baseline concentration of IgE and the IgG 4/IgG ratio. Conclusion:IgG 4-RDs are a group of diseases characterized by male predisposition; multiple organ involvement; a high eosinophil count; high IgE, IgG, IgG 1, and IgG 4 concentrations; and a low C3 concentration. Peripheral CD4 +T cells and Treg cells are also more abundant. The diseases can be controlled with glucocorticoids and immunosuppressive drugs in the majority of instances. The IgG 4 concentration after 6 months of treatment negatively correlates with the baseline complement C4 concentration and positively correlates with the IgE concentration and IgG 4/IgG ratio, which suggests that IgG 4/IgG, IgE, and complement should be closely monitored to evaluate disease activity and the efficacy of treatment in such patients.
2.Single hydrogen-methane breath test for the diagnosis of small intestinal bacterial growth
Huan HUANG ; Huizhen LI ; Yanrong WANG ; Yan SONG ; Bangmao WANG ; Hailong CAO ; Kui JIANG
Chinese Journal of Internal Medicine 2023;62(11):1335-1340
Objective:To investigate the diagnostic value of a single hydrogen-methane breath test (SHMBT) for small intestinal bacterial overgrowth (SIBO).Method:The current investigation was a cross-sectional study. Questionnaires and SHMBTs were administered to 162 patients with gastrointestinal symptoms (case group) and 69 healthy volunteers (control group). Differences in SHMBT results between the two groups were assessed,and cut-off values of CH 4 (methane) and H 2 (hydrogen) were analyzed via receiver operating characteristic (ROC) curves. Lastly,archived SHMBT data from 2 655 patients with gastrointestinal symptoms (validation set) were used to evaluate the diagnostic value of the SHMBT with respect to SIBO. The Chi-square test,the Mann-Whitney U test,Spearman′s Rank correlation analysis,and the Z test were used for statistical analysis. Results:Based on the international recommended diagnostic criteria for SIBO,which are fasting CH 4 ≥10 ppm (parts per million) or H 2 ≥20 ppm,the SHMBT-positive rate in the case group was significantly higher than that of control group (35.2% vs. 21.7%, χ2=4.08, P=0.043). Levels of CH 4 and H 2 were higher in the case group than in the control group [CH 4: 3(2,7) vs. 3(1,3) ppm, H 2: 11(4,22) vs. 10(5,15) ppm],and the difference in CH 4 levels was statistically significant ( Z=6.22, P=0.001). ROC curves were generated based on whether the subjects had gastrointestinal symptoms. The areas under the ROC curves were 0.633 for CH 4 alone,0.531 for H 2 alone, and 0.620 for CH 4 combined with H 2. The cut-off values were fasting CH 4≥4 ppm,fasting H 2≥13 ppm,and fasting CH 4 ≥5 ppm (or CH 4≥4 ppm and H 2≥24 ppm),respectively. Measuring CH 4 alone and CH 4 combined with H 2 was effective for determining the presence of gastrointestinal symptoms ( P<0.05). When CH 4 alone or CH 4 combined with H 2 were used as diagnostic indicators of SIBO, the respective SHMBT-positive rates in the validation set were 34.2% and 30.4%. These rates did not significantly differ from the SIBO-positive rate of 32.0% obtained via the international recommended diagnostic criteria ( P>0.05). The specificity of CH 4 alone was 79.9%,and the accuracy of CH 4 alone was 68.8%. The specificity of CH 4 combined with H 2 was 85.0%,and the accuracy of CH 4 combined with H 2 was 71.7%. Conclusion:Rapid one-time determination of CH 4 and H 2 in exhaled breath may a viable diagnostic method for SIBO, and using CH 4 combined with H 2 ( i.e.,fasting CH 4≥5 ppm, or CH 4 ≥4 ppm and H 2 ≥24 ppm) as cutoff values may be feasible.
3.Effect of depression on response to ursodeoxycholic acid and the occurrence of liver cirrhosis in patients with primary biliary cholangitis
Haifeng LIU ; Simin ZHOU ; Jiwen LI ; Jiangpeng LIU ; Bangmao WANG ; Lu ZHOU
Journal of Clinical Hepatology 2023;39(12):2817-2823
ObjectiveDepression is common in patients with primary biliary cholangitis (PBC), but the role of depression in disease progression remains unclear. This study aims to investigate the association between depression and treatment response and the impact of depression on liver cirrhosis in PBC patients. MethodsA retrospective analysis was performed for the clinical data of 141 patients with PBC who attended the outpatient service of autoimmune liver diseases in General Hospital of Tianjin Medical University from January 2018 to December 2020 and received standard ursodeoxycholic acid (UDCA) monotherapy for 1 year, and 170 healthy controls, matched for age and sex, who underwent physical examination in Physical Examination Center were enrolled as healthy control group. Patient Health Questionnaire-9 (PHQ-9) was used to evaluate depressive state in the patients with PBC and the healthy controls. The independent-samples t test was used for comparison of normally distributed continuous data between two groups, and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups; the chi-square test was used for comparison of categorical data between two groups. The binary logistic regression model and the decision tree model were used to analyze the influencing factors for liver cirrhosis in patients with PBC, as well as the influence of depression and the HLA-DRB1 gene on liver cirrhosis. The receiver operating characteristic (ROC) curve, the area under the ROC curve (AUC), and goodness of fit were used to evaluate model performance. All 13 variables were used to establish a classification and regression tree (CART) model, i.e., age, sex, PHQ-9 score, the DRB1*03∶01 gene, and the serum levels of alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase (ALP), gamma-glutamyl transpeptidase (GGT), total bilirubin, immunoglobulin G, immunoglobulin M (IgM), C3, and C4. The indications including AUC, sensitivity, and specificity were used to evaluate the performance of CART model in the model cohort. ResultsCompared with the normal control group, the PBC group had a significantly higher proportion of the patients with depression (53.9% vs 15.3%, χ2=57.836, P<0.001). Compared with the PBC patients without depression, the PBC patients with depression had a significantly poorer response to UDCA treatment (χ2=7.549, P=0.006) and significant increases in the serum levels of ALP (Z=-2.157, P=0.031), GGT (Z=-2.180, P=0.029), and IgM (Z=-2.000, P=0.046). Compared with the PBC patients without depression, the PBC patients carrying the HLA-DRB1*03∶01 allele had a significant increase in the risk of liver cirrhosis (P<0.001). The binary logistic regression model analysis showed that PHQ-9 score (OR=1.148, 95%CI: 1.050 — 1.255, P=0.002), the HLA-DRB1*03∶01 gene (OR=5.150, 95%CI: 1.362 — 19.478, P=0.016), age (OR=1.057, 95%CI: 1.009 — 1.106, P=0.018), and serum ALP level (OR=1.009, 95%CI: 1.001 — 1.017, P=0.020) were independent risk factors for liver cirrhosis in patients with PBC. The decision tree analysis showed that PHQ-9 score ≥3.5 was also a risk factor for liver cirrhosis in PBC patients. ConclusionDepression is associated with poor treatment response in patients with PBC, and it is an independent risk factor for liver cirrhosis in patients with PBC. This study highlights the important clinical significance of the identification and early management of depressive state in patients with PBC.
4.Exploring the links between gut microbiome changes and irritable bowel syndrome in Han populations in the Tibetan Plateau.
Xingguang ZHANG ; Wei XU ; Weilong ZHONG ; Wencheng ZHANG ; Cheng YANG ; Lisa DUAN ; Haiyan NIU ; Yanmei DONG ; Taotao LIU ; Shihai XIA ; Bangmao WANG
Journal of Zhejiang University. Science. B 2023;():1-16
The gut microbiome shows changes under a plateau environment, while the disbalance of intestinal microbiota plays an important role in the pathogenesis of irritable bowel syndrome (IBS); however, the relationship between the two remains unexplored. In this work, we followed up a healthy cohort for up to a year before and after living in a plateau environment and performed 16S ribosomal RNA (rRNA) sequencing analysis of their fecal samples. Through evaluating the participants' clinical symptoms, combined with an IBS questionnaire, we screened the IBS sub-population in our cohort. The sequencing results showed that a high-altitude environment could lead to changes in the diversity and composition of gut flora. In addition, we found that the longer the time volunteers spent in the plateau environment, the more similar their gut microbiota composition and abundance became compared to those before entering the plateau, and IBS symptoms were significantly alleviated. Therefore, we speculated that the plateau may be a special environment that induces IBS. The taxonomic units g_Alistipes, g_Oscillospira, and s_Ruminococcus_torques, which had been proved to play important roles in IBS pathogenesis, were also abundant in the IBS cohort at high altitudes. Overall, the disbalance of gut microbiota induced by the plateau environment contributed to the high frequency of IBS and the psychosocial abnormalities associated with IBS. Our results prompt further research to elucidate the relevant mechanism.
5.A comparative study of gastrointestinal gas volume in patients with functional dyspepsia based on abdominal computed tomography and gas volume score
Bin WANG ; Wei ZHAO ; Lili ZHANG ; Hong JIN ; Xin CHEN ; Yi WANG ; Bangmao WANG
Chinese Journal of Digestion 2023;43(10):677-682
Objective:To compare the difference between gas volume score (GVS) method and combination of computed tomography (CT) image and calculation formula method (hereinafter referred to as CT method) in the detection of gastrointestinal gas volume in functional dyspepsia (FD) patients.Methods:From December 1, 2021 to June 30, 2022, 27 FD patients (FD group) who visited the Department of Gastroenterology and Hepatology of Tianjin Medical University General Hospital were enrolled. At the same period, 30 healthy controls were selected from the database of check-up center as the healthy control group. All the participants of the two groups underwent erect plain abdominal X-ray and abdominal CT scan. The GVS and CT methods were used to calculate and compare gastrointestinal gas volume between two groups of patients, as well as patients with FD subtypes (postprandial distress syndrome (PDS), epigastric pain syndrome (EPS), and PDS overlapping with EPS). Independent sample t-test and one-way ANOVA were used for statistical analysis. Results:Based on the GVS method, the gas volume of gastric cavity, small intestine and colorectum of FD group were 0.04±0.01, 0.06±0.01 and 0.06±0.01, respectively; and those of the healthy control group were 0.04±0.01, 0.05±0.01 and 0.05±0.01, respectively. The gas volume of small intestine and colorectum of FD group were higher than those of the healthy control group, and the differences were statistically significant( t=3.48 and 4.40, P=0.001 and <0.001). The gas volume of gastric cavity, small intestine and colorectum of FD patients with subtypes of PDS, EPS, and PDS overlapping with EPS were 0.04±0.01, 0.04±0.01 and 0.05±0.00, 0.06±0.01, 0.06±0.01 and 0.05±0.00, and 0.06±0.01, 0.06±0.01 and 0.06±0.01, respectively. There were no significant difference in the gas volume of gastric cavity, small intestine and colorectum among different subtypes ( all P>0.05 ). Based on the CT method, the gas volume of gastric cavity, small intestine and colorectum of FD group were (17 090.89±4 437.40) mm 3, (32 597.53±7 865.86) mm 3 and (49 010.20±12 972.42) mm 3, respectively; and those of the healthy control group were (13 424.43±5 211.86) mm 3, (33 567.93±9 157.23) mm 3 and (39 036.22±6 343.27) mm 3, respectively. The gas volume of gastric cavity and colorectum of FD group were higher than those of the healthy control group, and the differences were statistically significant( t=2.84 and 3.75, P=0.006 and 0.001). The gas volume of gastric cavity, small intestine and colorectum of FD patients with subtypes of PDS, EPS, and PDS overlapping with EPS were (18 464.03±4 088.57) mm 3, (14 560.97±3 771.26) mm 3 and (16 806.17±4 299.60) mm 3, (31 820.79±7 022.77) mm 3, (30 604.84±8 343.10) mm 3 and (37 140.05±8 276.58) mm 3, and (47 447.66±14 047.00) mm 3, (49 645.73±9 527.73) mm 3 and (51 181.96±16 836.97) mm 3, respectively. The gastric gas volume of FD patients with subtype of PDS was higher than that of FD patients with subtype of EPS, and the difference was statistically significant ( t=2.24, P=0.038). Conclusions:The volume of gastrointestinal gas of FD patients is higher than that of healthy controls. Gas accumulation in the gastric cavity of FD patients with subtype of PDS is more significant than that of FD patients with subtype of EPS. The CT method may assist physicians in calculating the gas volume in the gastrointestinal tract (especially in stomach) of FD patients more accurately.
6.Expert consensus on recombinant B subunit/inactivated whole-cell cholera vaccine in preventing infectious diarrhea of enterotoxigenic Escherichia coli
Chai JI ; Yu HU ; Mingyan LI ; Yan LIU ; Yuyang XU ; Hua YU ; Jianyong SHEN ; Jingan LOU ; Wei ZHOU ; Jie HU ; Zhiying YIN ; Jingjiao WEI ; Junfen LIN ; Zhenyu SHEN ; Ziping MIAO ; Baodong LI ; Jiabing WU ; Xiaoyuan LI ; Hongmei XU ; Jianming OU ; Qi LI ; Jun XIANG ; Chen DONG ; Haihua YI ; Changjun BAO ; Shicheng GUO ; Shaohong YAN ; Lili LIU ; Zengqiang KOU ; Shaoying CHANG ; Shaobai ZHANG ; Xiang GUO ; Xiaoping ZHU ; Ying ZHANG ; Bangmao WANG ; Shuguang CAO ; Peisheng WANG ; Zhixian ZHAO ; Da WANG ; Enfu CHEN
Chinese Journal of Clinical Infectious Diseases 2023;16(6):420-426
Enterotoxigenic Escherichia coli(ETEC)infection can induce watery diarrhea,leading to dehydration,electrolyte disturbance,and even death in severe cases. Recombinant B subunit/inactivated whole-cell cholera(rBS/WC)vaccine is effective in preventing ETEC infectious diarrhea. On the basis of the latest evidence on etiology and epidemiology of ETEC,as well as the effectiveness,safety,and health economics of rBS/WC vaccine,National Clinical Research Center for Child Health(The Children’s Hospital,Zhejiang University School of Medicine)and Zhejiang Provincial Center for Disease Control and Prevention invited experts to develop expert consensus on rBS/WC vaccine in prevention of ETEC infectious diarrhea. It aims to provide the clinicians and vaccination professionals with guidelines on using rBS/WC vaccine to reduce the incidence of ETEC infectious diarrhea.
7.Oral sulfate solution versus polyethylene glycol for colonoscopy bowel preparation: a randomized controlled study in phase Ⅲ
Ye ZONG ; Fandong MENG ; Yongdong WU ; Bangmao WANG ; Xizhong SHEN ; Yi CUI ; Guoxin ZHANG ; Aiming YANG ; De'an TIAN ; Jianting CAI ; Huahong WANG ; Shihua CUI ; Min CUI ; Shutian ZHANG
Chinese Journal of Digestive Endoscopy 2022;39(4):261-266
Objective:To compare the efficacy of oral sulfate solution (OSS) and polyethylene glycol (PEG) electrolyte powder for colonoscopy bowel preparation.Methods:A total of 283 randomized patients from 9 centers in China taking OSS ( n=143) or PEG ( n=140) using two-day split bowel preparation regimen received colonoscopy and assessment. The primary index was the bowel preparation success rate [global Boston bowel preparation scale (BBPS)≥ 6 by independent assessment center]. Secondary indices included BBPS global and segmental scores, investigator satisfaction (5-point Likert scale) with the quality of bowel preparation, patient satisfaction assessed by questionnaires, and patient tolerance assessed by Sharma scale. Compliance and safety were compared between the two groups. Results:The bowel preparation success rates were 100.0% for OSS and 99.3% for PEG [adjusted difference 0.7% (95% CI: -5.3% - 6.7%), P<0.001 for non-inferiority]. The BBPS global score in OSS group was significantly higher than that in PEG group (8.1 VS 7.7, P<0.001). The segment BBPS scores were also higher in OSS group than those in PEG group for all 3 segments (right colon: 2.4 VS 2.3, P=0.002; transverse colon: 2.8 VS 2.7, P=0.018; left colon: 2.8 VS 2.7, P=0.007). Investigator Likert score in the OSS group was significantly higher than that in the PEG group (2.6 VS 2.3, P<0.001). There was no significant difference in compliance between OSS and PEG, except for the second dose (90.9% VS 82.6%, P=0.039). There was no significant difference in patient satisfaction, Sharma score or proportion of patients with tolerance-related symptoms between the two groups. Safety was comparable between the two groups, and all adverse events were mild to moderate. Conclusion:OSS has comparable efficacy with PEG, with higher BBPS scores in all segments, better investigator satisfaction, better compliance in split dose, and comparable patient tolerance and safety.
8.Inhibition of receptor-interacting protein 3 improves experimental autoimmune hepatitis
Man LIU ; Hongxia ZHANG ; Lu ZHANG ; Jingwen ZHAO ; Lu ZHOU ; Bangmao WANG
Chinese Journal of Hepatology 2022;30(11):1231-1236
Objective:To evaluate the potential of receptor-interacting protein 3 (RIP3) as a therapeutic target for autoimmune hepatitis (AIH).Methods:Immunofluorescence assay was used to observe the activated expression levels of RIP3 and its downstream signal mixed lineage protein kinase domain-like protein (MLKL) in the liver tissues of patients with AIH and hepatic cyst. Concanavalin A (ConA) was injected into the tail vein to induce acute immune-mediated hepatitis in mice. Intervention was performed by intraperitoneal injection of RIP3 inhibitor GSK872 or solvent carrier. Peripheral blood and liver tissues were collected. Serum transaminases level, qPCR and flow cytometry were analyzed. The intergroup comparison was performed with an independent sample t-test. Results:The expression level of p-RIP3 (the activated forms of RIP3) and phosphorylated p-MLKL (MLKL after phosphorylation) downstream signal were significantly higher in the liver tissue of AIH patients than those of controls. Compared with the control group, the expression levels of RIP3 and MLKL mRNA were significantly increased in the liver tissue of AIH patients (relative expression levels 3.28±0.29 vs. 0.98±0.09, 4.55±0.51 vs. 1.06±0.11), and the differences were statistically significant ( t=6.71 and 6.77, respectively, and P<0.01). The expression levels of RIP3 and MLKL mRNA were significantly higher in the mice liver tissue of ConA-induced immune hepatitis than those in the control group (relative expression levels 2.35±0.09 vs. 0.89±0.11,2.77±0.22 vs. 0.73±0.16, t=10.4,6.33, P<0.01). RIP3 inhibitor GSK872 had significantly attenuated ConA-induced immune liver injury and inhibited the expression of tumor necrosis factor-α, interleukin-6, interleukin-1β and NLRP3 in liver. Compared with the control group, the proportions of CD45 +F4/80 + macrophages, CD4 + IL-17 + Th17 cells, CD4 + CD25 + regulatory T (Treg) cells and CD11b + Gr-1 + myeloid derived suppressor cells (MDSCs) were significantly increased in the liver of ConA + Vehicle group. Compared with ConA + Vehicle group, the proportion of CD45 +F4/80 + macrophages and CD4 + IL-17 + Th17 cells were significantly decreased, while the proportion of CD4 + CD25 +Treg cells and CD11b + Gr-1 + MDSCs with immunomodulatory functions were significantly increased in mice liver of ConA+GSK872 group. Conclusion:AIH patients and ConA-induced immune hepatitis mice have activated RIP3 signal in liver tissues. Inhibition of RIP3 reduces the expression and proportion of proinflammatory factors and cells, and promotes the accumulation of CD4 + CD25 + Treg cells and CD11b + Gr-1 + MDSCs with immunomodulatory functions in the liver of mice with immune hepatitis, thereby alleviating liver inflammation and injury. Therefore, the inhibition of RIP3 is expected to be a new approach for the treatment of AIH.
9.Effect of dietary intervention on blood glucose in colonoscopy patients during intestinal preparation: a meta-analysis
Li LIANG ; Weilong ZHONG ; Bangmao WANG
Chinese Journal of Digestive Endoscopy 2021;38(4):310-314
Objective:To systematically evaluate the effect of dietary intervention on blood glucose during intestinal preparation for colonoscopy.Methods:Randomized controlled trials (RCTs) on the effects of different interventions on blood glucose during bowel preparation in colonoscopy patients were retrieved on PubMed, CNKI, and WanFang, and RevMan 5.3 software was applied for meta-analysis based on the intervention groups.Results:A total of 10 RCTs in 8 papers, including 3 345 patients, 1 603 patients in the control group and 1 742 patients in the intervention group, were reviewed. Meta-analysis results showed that 10 studies reported incidence of hypoglycemia and interventions during intestinal preparation, and heterogeneity among researches was statistical ( P<0.000 01, I2=80%). Under random effects model combined with effect quantity, compared with the control group, dietary intervention could significantly reduce the incidence of hypoglycemia during bowel preparation, the difference was significant ( OR=0.19, 95% CI: 0.09-0.40, P<0.05). Conclusion:Dietary intervention during intestinal preparation for colonoscopy can prevent the occurrence of hypoglycemia and avoid relative adverse reactions and parasympathetic nerve activity. However, due to the limited number and quality of included researches, the above conclusion need to be verified by more high-quality RCTs.
10.Receptor interaction protein 3 mediated the recruitment of hepatic monocytes/macrophages in autoimmune hepatitis
Man LIU ; Hongxia ZHANG ; Lu ZHOU ; Bangmao WANG
Chinese Journal of Digestion 2021;41(1):35-42
Objective:To explore the role of receptor-interaction protein 3 (RIP3) in regulating the infiltration of monocytes/macrophages into the liver in autoimmune hepatitis (AIH).Methods:From January to June in 2018, at Department of Gastroenterology and Hepatology, Tianjin Medical University General Hospital, 10 AIH patients who underwent liver biopsy were enrolled, and at the same time, 5 age and gender matched individuals with normal liver function and hepatic cyst were selected as control. The infiltration of monocytes/macrophages in the liver tissues was observed by immunofluorescence detection in the patients with AIH and controls. Raw264.7 macrophages were divided into control group, lipopolysaccharide group and lipopolysaccharide+ RIP3 inhibitor GSK872 (GSK872) group. The expression of RIP3, mixed lineage kinase domain like pseudokinase ( MLKL), tumor necrosis factor ( TNF)- α, interleukin ( IL)-6, IL-1 β, nod-like receptor protein 3 ( NLRP3), CC motif chemokine ligand ( CCL)2 and CCL5 at mRNA levels were detected by quantitative polymerase chain reaction (qPCR). Raw264.7 macrophages were also divided into control group, lipopolysaccharide group and lipopolysaccharide + dexamethasone group. The relative expression of TNF- α, NLRP3, RIP3 and MLKL at mRNA level in macrophage were detected by qPCR. Twenty-four 6-week-old female C57BL/6 mice were chosen to establish AIH mice model and were randomly divided into control group, concanavalin A (ConA) group, ConA+ dexamethasone group and ConA+ GSK872 group (6 mice in each group). After the mice were executed, the peripheral blood and liver tissues were collected. The histopathology of mice liver were observed and the serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were measured. The expression of CCL2 and CC motif chemokine receptor 2 ( CCR2) at mRNA level were detected by qPCR. The proportion of macrophages in mice livers were analyzed by flow cytometry. The independent sample t test and one-way analysis of variance were performed for statistical analysis. Results:The percentages of CD68 positive macrophages and MAC387 positive infiltrated mononuclear macrophages in livers of AIH patients were both higher than those of controls ((0.84±0.21)% vs. (0.09±0.03)%, (0.79±0.13)% vs. (0.03±0.01)%), and the differences were statistically significant ( t=3.00 and 4.84; all P<0.05). The expression of RIP3, MLKL, TNF- α, IL-6, IL-1 β, NLRP3, CCL2 and CCL5 at mRNA level of lipopolysaccharide group were all higher than those of control group and lipopolysaccharide+ GSK872 group (1.64±0.16 vs. 1.07±0.07 and 0.63±0.11; 10.45±1.37 vs. 1.10±0.33 and 1.51±0.63; 5.43±0.59 vs. 0.94±0.06 and 2.59±0.45; 204.20±30.73 vs. 1.26 ±0.19 and 111.40±11.62; 20 848.00±362.00 vs. 1.09 ±0.26 and 10 940.00±566.60; 7.47±1.17 vs. 1.09±0.09 and 3.79±0.89; 68.03±5.15 vs. 1.14±0.19 and 14.09±2.62; 5 935.12±96.20 vs. 1.43±0.46 and 673.50±49.10), and the differences were all statistically significant ( t=3.11, 5.21, 6.65, 6.55, 7.57, 3.96, 6.60, 3.06, 8.83, 4.08, 5.46, 2.56, 12.97, 10.16, 25.34 and 14.99; all P<0.05). The expression of TNF- α, NLRP3, RIP3 and MLKL at mRNA level of lipopolysaccharide group were all higher than those of control group and lipopolysaccharide+ dexamethasone group (8.85±1.43 vs. 1.44±0.43 and 3.63±0.63; 6.42±0.86 vs. 0.99±0.12 and 2.07±0.17; 1.72±0.21 vs. 0.93±0.09 and 0.43±0.07; 6.87±0.85 vs. 1.62±0.31 and 1.41±0.29), and the differences were all statistically significant ( t=4.95, 3.33, 6.24, 4.95, 3.04, 5.11, 5.77 and 6.07, all P<0.05). The mice liver of ConA group showed obviously inflammatory cells infiltration and hepatocytes necrosis. The serum ALT and AST levels of ConA group were both higher than those of control group, ConA+ dexamethasone group and ConA+ GSK872 group ((2 569.00±45.44) U/L vs. (49.38±9.07), (103.00±14.07) and (759.30±34.99) U/L; (3 335.00±88.79) U/L vs. (108.50±18.10), (460.00±97.40) and (1 573.85±36.06) U/L), the serum ALT and AST levels of ConA+ dexamethasone group were both lower than those of ConA+ GSK872 group, and the differences were all statistically significant ( t=5.54, 5.42, 3.90, 4.63, 4.16, 3.79, 6.70 and 2.71; all P<0.05). The expression of CCL2 and CCR2 at mRNA levels in mice liver of ConA group were both higher than those of control group, ConA+ dexamethasone group and ConA+ GSK872 group (92.64±10.57 vs. 0.78±0.15, 5.64±1.00 and 9.47±2.06; 5.73±0.39 vs. 0.98±0.22, 2.18±0.22 and 2.98±0.33), and the differences were all statistically significant ( t=7.66, 7.24, 5.87, 8.71, 8.58 and 5.45; all P <0.01). The proportion of CD45 + CD11b + F4/80 + total macrophages and CD45 + CD11b hiF4/80 lo infiltrated macrophages in mice livers of ConA group were both higher than those of control group, ConA+ dexamethasone group and ConA+ GSK872 group (0.86±0.02 vs. 0.73±0.03, 0.68±0.02 and 0.72±0.03; 0.56±0.02 vs. 0.08±0.02, 0.11±0.01 and 0.08±0.01), however the proportion of CD45 + CD11b loF4/80 hi liver macrophages (Kupffer cells) was lower than those that of control group, ConA+ dexamethasone group and ConA+ GSK872 group (0.24±0.03 vs. 0.58±0.04, 0.52±0.07 and 0.56±0.07), and the differences were all statistically significant ( t=4.27, 5.90, 3.89, 18.70, 19.87, 20.52, 7.35, 3.82 and 3.87, all P<0.05). Conclusions:The number of macrophages incread in the livers of AIH patients. RIP3 signaling mediates the migration of monocytes/macrophages infiltration in immune hepatitis, which may be a potential therapeutic target for AIH.

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