Objective To investigate the effects of interleukin(IL)-34 on the polarization and migration ability of macrophages derived from human peripheral blood monocytes.Methods The CD14+monocytes were isolated from human peripheral blood monocytes by immunomagnetic bead sorting,and the purity of CD14+monocytes was detected by flow cytometry.The CD14+monocytes were divided into M1 type group,IL-34-M1 type group,M2 type group and IL-34-M2 type group.The cells in the M1 type group and IL-34-M1 type group were added granulocyte-macrophage colony stimulating factor(GM-CSF)to induce for 5 days,and half of the fluid was changed,and then the interferon-γ,lipopolysaccharides,IL-6 and GM-CSF were added for another 4-day induction;the cells in the M2 type group and IL-34-M2 type group were added macrophage colony stimulating factor(M-CSF)to induce for 5 days,and half of the fluid was changed,and M-CSF,IL-4,IL-6,and IL-13 were added for another 4-day induction.The cells in IL-34-M1 group and IL-34-M2 group were co-induced with IL-34 at the beginning of induction and on the 5th day of induction.On the 9th day of induction,the proportion of CD14+CD86+cells(M1 type macrophages)and CD14+CD163+cells(M2 type macrophages)in each group was detected by flow cytometry,and the migration ability of cells in the M2 type group and IL-34-M2 type group was detected by the Transwell chamber experiments.Results High purity CD14+monocytes were obtained through magnetic bead sorting,with a CD14 positive rate of(96.77± 2.72)％,which could be used for macrophage induction.The proportion of CD14+CD86+cells in the M1 type group and IL-34-M1 type group was(43.20±7.59)％and(27.87±2.06)％,respectively.The proportion of CD14+CD163+cells in the M2 type group and IL-34-M2 type group was(47.70±4.49)％and(58.95±3.65)％,respectively;the proportion of CD14+CD86+cells in the IL-34-M1 type group was significantly lower than that in the M1 type group(P＜0.05),while the proportion of CD14+CD163+cells in the IL-34-M2 type group was significantly higher than that in the M2 type group(P＜0.05).The number of migrating cells of macrophages in the M2 type group and IL-34-M2 type group was 97.8±9.0 and 205.6±21.9,respectively;the number of migrating cells of macrophages in the IL-34-M2 type group was significantly higher than that in the M2 type group(P＜0.05).Conclusion IL-34 can inhibit the polarization of macrophages derived from human monocytes cells towards M1 type,promote the polarization of macrophages towards M2 type,and enhance the migration ability of M2 type macrophages.

The gastrointestinal tract is an extremely complex ecosystem,and a healthy and intact intestinal mucosal barrier is the basic defense against the translocation of harmful substances.The impaired function of the intestinal barrier in acute-on-chronic liver failure(ACLF)is further aggravated by the dysregulation of intestinal microecology,which leads to bacterial translocation and endotoxemia.Maintaining the normal function of the intestinal mucosal barrier is important for the treatment of ACLF.This paper reviews the function of the normal intestinal mucosal barrier,the relationship between the damaged intestinal barrier and ACLF,and the impact of the damaged intestinal barrier on ACLF progression.

Background: Hepatic encephalopathy (HE) is one of the main causes of death in patients with end-stage liver disease. Early evaluation of the condition of patients with liver cirrhosis complicated with HE is the key for improving the prognosis. Aims: To investigate the effects of controlling nutritional status (CONUT) and CTP score on predicting the short term prognosis of liver cirrhosis patients complicated with HE. Methods: The clinical data of 168 liver cirrhosis patients complicated with HE initially diagnosed in Nantong Third People's Hospital from January 2018 to December 2021 were enrolled and analyzed retrospectively. HE patients were divided into survival group and death group. Cox regression analysis was used to evaluate the risk factors affecting the prognosis. A new prediction model was established, and ROC curve was used to evaluate the values of different models for short⁃term prognosis of HE patients. Survival was evaluated by Kaplan⁃Meier method. Results: Compared with survival group, ALT, AST, TBIL, INR, white blood cell count, neutrophil were significantly increased in death group (P<0.05), while ALB, total cholesterol, serum Na, fibrinogen, lymphocyte were significantly decreased (P<0.05). CONUT (OR=1.499, 95% CI: 1.092⁃2.057, P=0.012), CTP score (OR=1.474, 95% CI: 1.178⁃1.844, P=0.001) were independent risk factors for 90⁃day mortality in liver cirrhosis patients complicated with HE. AUC of CTP, CONUT and CONUT⁃CTP scoring models were 0.90, 0.94 and 0.95, respectively. Kaplan⁃Meier analysis showed that the survival rate in CONUT⁃CTP≥7.50 group was significantly lower than that in <7.50 group (P<0.000 1). Conclusions: CONUT⁃CTP score has good predictive value for the prognosis of liver cirrhosis patients complicated with HE, CONUT⁃CTP ≥7.50 indicates poor prognosis.

Background: Interleukin-34 (IL-34) is an important immunomodulatory factor that plays a crucial role in a variety of autoimmune diseases. Aims: To investigate the expression of IL-34 in primary biliary cholangitis (PBC) and its influence on intrahepatic inflammation and bile duct damage. Methods: Liver tissues were obtained from 26 PBC patients and 10 hepatic hemangioma patients without PBC. Expression and localization of IL-34 were detected by immunohistochemistry. In animal experiment, Poly I:C intraperitoneal injection was used to construct PBC model in wild-type and IL-34-knockout C57BL/6 mice (WT-PBC group and IL-34KO-PBC group). Subsequently, the intrahepatic inflammation and bile duct damage were evaluated pathologically, and the expressions of IL-34 and associated cytokines in liver tissues were detected by real-time PCR and Western blotting. Results: Expression of IL-34 in liver tissues of PBC patients and PBC model mice was significantly higher as compared with those of the controls (all P<0.05). No morphological changes in hepatic pathological evaluation were observed in IL-34KO mice receiving intraperitoneal saline injection. In IL-34KO-PBC mice, the portal area inflammation and biliary epithelial cell damage were more severe than those in WT-PBC mice (all P<0.05). Expressions of proinflammatory cytokine interleukin-1β (IL-1β) and monocyte chemotactic protein-1 (MCP-1) in liver tissues of IL-34KO-PBC mice were significantly increased than those of WT-PBC mice, whereas expressions of antiinflammatory cytokine IL-10 and CD163, the surface marker of M2 macrophages, were significantly reduced (all P<0.05). Conclusions: IL-34 expression is increased in liver tissues of PBC patients and animals. It might reduce the portal area inflammation and bile duct damage via modulating cytokines expression and driving macrophages polarization to the M2 phenotype. IL-34 might act as a self-rescue factor which negatively regulates hepatic immune microenvironment and prevents disease progression.

Objective:To explore the role of macrophages in non-alcoholic steatohepatitis (NASH) in order to provide directions for the therapeutic target of metabolic liver disease.Methods:Twenty C57BL/6 wild-type male mice at 6-8 weeks were randomly divided into two groups: 5 in the control group, methionine-and choline-deficient diet (MCD); 15 in the experimental group, MCD diet + intraperitoneal injection of disodium chlorophosphonate liposomes (to clear macrophages). Mice were fed for 4 weeks to establish NASH model. Blood, liver and spleen were collected to analyze the body mass index, liver index, spleen index, and serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels. Non-alcoholic steatosis (NAS) activity score was evaluated by HE and Oil Red O staining. The relative expression level of F4/80 mRNA was compared by RT-PCR. Data comparison between groups was analyzed by t-test.Results:NASH model was successfully established by feeding the mice with MCD for four week. The expression of F4/80 mRNA ( t = 4.167, P < 0.01), hepatic steatosis ( t = 10.70, P < 0.05), interlobular inflammatory infiltration ( t = 3.08, P < 0.05), and NAS score were decreased ( t = 8.06, P < 0.05) in the experimental group. At the same time, ALT level [(817.00 ± 128.90) U/L vs. (231.20 ± 36.28) U/L, t = 5.71, P < 0.01], AST level [(1 211.00 ± 248.90) U/L vs. (505.30 ± 88.20) U/L, t = 3.32, P < 0.01] was decreased significantly. However, the spleen volume and spleen index of the experimental group were larger (0.24 ± 0.01 and 0.32 ± 0.02, t = 2.41, P < 0.05), and there was no significant effect on liver ballooning, body mass index and liver index. Conclusion:In NASH, phosphonate can consume macrophages to inhibit liver inflammation and protect the damaged liver.

Thirty mice were used to establish a sepsis model with cecal ligation and puncture. 15 mg/kg methylene blue or isotonic saline were injected intraperitoneally to observe liver tissue pathological changes. Changes in macrophage frequency and expressional condition of M1 and M2-type hepatic inflammatory factors were detected. After LPS stimulation, the expression level of macrophage inflammatory factor were detected. The results showed that the pathological liver injury was significantly reduced in the MB mice group ( P < 0.05), and the frequency of liver macrophage was not statistically significantly different ( P > 0.05). MB elevation had promoted the expression of M2-type hepatic inflammatory factor ( P < 0.05) and macrophage inflammatory factor ( P < 0.05). MB can play a role in preventing septic liver injury by inducing macrophages polarization to M2-type.

Objective:To investigate the correlation between the expression of programmed death ligand-1 (PD-L1) and clinicopathological parameters and prognosis in hepatocellular carcinoma (HCC) tissues.Methods:From January 2008 to December 2016, 344 HCC patients underwent surgery in Nantong Third Hospital Affiliated to Nantong University were enrolled. The expression of PD-L1 in paraffin HCC tissues was detected by tissue microarray and immunohistochemistry. The correlation between the expression of PD-L1 in tumor cells, tumor-infiltrating immune cells and the clinicopathological parameters and prognosis of HCC patients were analyzed. And the related factors affecting the prognosis of patients were explored. Chi-square test, log-rank test and univariate and multivariate Cox regression analysis were used for statistical analysis.Results:Positive PD-L1 located in the membrane and/or cytoplasm of HCC tumor cells and tumor-infiltrating immune cells. The positive rate of PD-L1 expression in tumor cells was 21.8%(75/344). The expression of PD-L1 in tumor cells was related to histological grade and microvascular invasion, the positive rates of PD-L1 expression of patients with histological gradeⅠ, Ⅱ and Ⅲ were 7.7% (2/26), 16.5% (19/115) and 26.6% (54/203), respectively, the positive rates of PD-L1 expression of patients with or without microvascular invasion were 29.3% (34/116) and 18.0% (41/228), respectively, and the differences were statistically significant ( χ2=7.659 and 5.787, P=0.022 and 0.016). The positive expression rate of PD-L1 in tumor-infiltrating immune cells was 47.1% (162/344). The expression of PD-L1 in tumor-infiltrating immune cells was related with microvascular invasion, the positive rates of PD-L1 expression in patients with or without microvascular invasion were 56.9% (66/116) and 42.1% (96/228), respectively, and the differences were statistically significant ( χ2=6.751, P=0.009). The median survival time of patients with negative expression of PD-L1 in HCC tumor cells was 61 months (30 months, 92 months), while that of patients with positive PD-L1 expression was 16 months(6 months, 44 months), and the difference was statistically significant ( χ2=55.722, P<0.01). The expression of PD-L1 in HCC tumor cells was an independent risk factor affecting overall survival time (hazard ratio=3.090, P<0.01). Conclusions:The expression of PD-L1 in HCC tumor cells may be related to the malignancy and invasion of HCC, and may be a potential risk factor for prognosis.

INTRODUCTION: Paraquat (PQ) intoxication is frequently associated with a high mortality rate. No specific treatment has been shown to reduce mortality in victims within the first 72 hours. We investigated the protective effects of rapamycin (Rapa) against PQ-induced toxicity in a zebrafish model. METHODS: To determine the maximum nonlethal concentration (MNLC) and lethal concentration 50 (LC50) of Rapa, zebrafish were treated at 2-5 days post fertilisation (dpf) and their mortality was recorded every 24 hours. At 5 dpf, the zebrafish were treated with PQ 100 µg/mL or PQ+Rapa (MNLC, 1/3 MNLC or 1/9 MNLC) for 72 hours, and the rate of survival was recorded every 24 hours. Reverse transcription-polymerase chain reaction was used to test the signalling pathway of mTOR (mammalian target of rapamycin). RESULTS: MNLC and LC50 of Rapa were determined to be 6.7 µg/mL and 28.9 µg/mL, respectively. At 48 hours, the PQ+Rapa groups had much lower mortality than the PQ group. The rates of survival of the PQ+Rapa groups were 43.33% (MNLC), 53.89% (1/3 MNLC) and 44.45% (1/9 MLNC), as compared to 19.45% in the PQ group, with the 1/3 MNLC group showing the highest rate of survival (p < 0.001). atg1 was slightly activated in the PQ group. In the PQ+Rapa groups, the expression of atg1 was markedly increased, suggesting strengthening of the autophagy process. CONCLUSION Rapa can increase the rate of survival of PQ-intoxicated zebrafish by inhibiting mTOR complex 1 and activating autophagy. Rapa could be an alternative first-line drug in the treatment of PQ poisoning.

Objective: To probe into the mechanism and interventional effects of silybin-phospholipid complex on amiodarone-induced steatosis in mice. Methods: Eight-week-old male C57BL/6 mice were divided into three groups (5 mice in each group): a control group (WT) with normal diet, a model group with amiodarone 150mg/kg/d by oral gavage (AM), and an intervention group on amiodarone 150mg/kg/d combined with silybin-phospholipid complex(AM+SILIPHOS. All mice were fed their assigned diet for one week. Then, one week later, serum alanine aminotransferase, aspartate aminotransferase, triglyceride, total cholesterol and high-density lipoprotein were detected of each group. A liver pathological change was observed by oil red O and H&E staining. Ultrastructural pathological changes of hepatocytes were observed to evaluate the intervention effect by transmission electron microscopy. RT-q PCR was used to detect the expression of peroxisome proliferator-activated receptor alpha and its regulated lipid metabolism genes CPTI, CPTII, Acot1, Acot2, ACOX, Cyp4a10 and Cyp4a14 in liver tissues. Intra-group comparison was done by paired t-test. One-way ANOVA was used for comparison between groups and semi-quantitative data were tested using Mann-Whitney U test. Results: Oil Red O and H&E staining results of liver tissue in the intervention group showed that intrahepatic steatosis was significantly reduced when compared to model group. Transmission electron microscopy showed that the model group had pyknotic nuclei, mitochondrial swelling, structural damage, and lysosomal degradation whereas the intervention group had hepatic nucleus without pyknosis, reduced mitochondrial swelling and slight structural damage than that of model group. RT-q PCR results showed that the expression of peroxisome proliferator-activated receptor alpha, CPTI, CPTII, Acot1, Acot2, ACOX, Cyp4a10 and Cyp4a14 were increased in the model group but the expression of CPTI, Cyp4a14, Acot1 and peroxisome proliferator-activated receptor alpha were decreased in the intervention group (P < 0.05). Conclusion Silybin-phospholipid complex can alleviate amiodarone-induced steatosis, and its mechanism may play a role in protecting mitochondrial function and regulating fatty acid metabolism. Thus, silybin-phospholipid complex has potential intervention effect on amiodarone-induced fatty liver.

Drug-induced liver injury ( DILI) and autoimmune hepatitis ( AIH) have many similar clinical and histological manifestations , which brings difficulties to clinicians in differential diagnosis .This article elaborates on the association between DILI and AIH and their simi-larities and differences in clinical and histological manifestations , in order to help clinicians with the differential diagnosis of DILI and AIH . This article reviews the selection of therapeutic regimens for DILI and AIH and introduces how to select therapeutic strategies based on patient conditions and make a definite diagnosis when there are difficulties in differential diagnosis .