[Comparison of serum GP73 and p62 measurement to predict short-term prognosis in patients with HBV-related acute-on-chronic liver failure].

Objective: To analyze the clinical value and predictive difference of serum Golgi protein 73 (GP73) and serum autophagy-related protein p62 levels in the short-term prognosis of patients with hepatitis B virus-related acute-on-chronic liver failure (ACLF). Methods: Clinical data of admitted cases to our hospital from October 2018 to April 2020 were retrospectively analyzed. Simultaneously, there were 32 cases with HBV-related ACLF in group A, 65 cases with hepatitis B virus-related cirrhosis in group B and C (Child-Pugh Class A, 34 cases as B group, and Child-Pugh B/C class, 31 cases as group C), and another 30 healthy subjects served as the control group (group D). The serum GP73 and p62 levels of the four selected groups were measured. ACLF group patients were followed up for 3 months to analyze the prognosis of the patients. The serum GP73 and p62 levels of patients who died and survived during hospitalization were compared. The data were analyzed by one-way analysis of variance, independent sample t-test, and Pearson's correlation analysis. Receiver operating characteristic curve (ROC) was used to analyze the predictive value of GP73 and p62 levels in surviving patients. Results: GP73 levels in the four groups A, B, C and D were (284.30 ± 70.55) ng/ml, (125.33 ± 20.57) ng/ml, (159.82 ± 31.20) ng/ml, and (45.46 ± 10.22) ng/ml, respectively. The p62 levels were (1.30 ± 0.35) ng/ml, (2.88 ± 0.58) ng/ml, (2.02 ± 0.545) ng/ml, and (4.68 ± 1.03) ng/ml, respectively. GP73 detection value was significantly higher in group A than the other three groups (P < 0.05). Group D had significantly lower value than the other three groups (P < 0.05), and group C had significantly higher value than group B (P < 0.05). The detection value of p62 in group A was significantly lower than the other three groups (P < 0.05). Group D had significantly higher value than the other three groups (P < 0.05), and group B had slightly higher value than group C, and the differences were statistically significant (P < 0.05). There was a negative correlation between GP73 and p62 (r = -0.695, P < 0.001). Survived patients GP73 level in the ACLF group was significantly lower than dead patients [(212.17 ± 22.47) ng/ml and (340.08 ± 32.91) ng/ml, t = 12.493, P < 0.05], and p62 level was significantly higher than dead patients [(1.46 ± 0.28) ng/ml and (1.18 ± 0.35) ng/ml, t = 2.445, P < 0.05]. According to the ROC curve analysis results, the area under the curve (AUC) of GP73 was 0.865, the AUC of p62 was 0.750, and the combined AUC of the both was 0.968. Conclusion: Both GP73 and p62 have a certain predictive value for the short-term prognosis of HBV-related ACLF patients, but the combination of the two indicators has a higher predictive value.

Eicosapentaenoic acid reduces inflammation and apoptosis by SREBP1/TLR4/MYD88.

AIM: Podocytes dysfunction including the cell integrity, apoptosis and inflammation plays crucial role in diabetic nephropathy. Current exploration evaluated the protective role of eicosapentaenoic acid (EPA) in high glucose-treated podocytes and the underlying mechanisms. METHOD: MPC5 cell were stimulated by high glucose or treated by EPA of different concentrations. CCK8 assay was utilized to assess MPC5 cell viability, flow cytometry analyzed cell apoptosis. RESULTS: Data showed that EPA prominently alleviated the high glucose-induced apoptosis and inflammation. Besides, the disruption of the podocytes structure certifying by podocin and synaptopodin induced by hyperglycemia was hindered by EPA administration. In addition, overexpression of the sterol regulatory element-binding protein-1 (SREBP-1) reversed the protective effects of EPA in high glucose-treated podocytes. EPA inhibits the SREBP-1/TLR4/MYD88 signaling in high glucose treated cells. CONCLUSIONS: This study suggests that EPA protects against podocytes dysfunction by regulating SREBP-1 and these findings provide a better understanding for diabetic nephropathy and a novel therapeutic strategy (Fig. 7, Ref. 24).

[Post-marketing multi-center safety surveillance of inactivated enterovirus A71 vaccine (Vero cell)].

Objective: To evaluate the post-marketing safety profiles of the inactivated enterovirus type 71 (EV-A71) vaccine (Vero cell) after routine inoculation. Methods: Eleven cities of Zhejiang Province, Fengtai district of Beijing, Qinnan district, two counties as Pingle and Pingguo of Guangxi Zhuang Autonomous Region, and Dongtai city of Jiangsu Province were selected as the field sites. A total of 45 239 subjects were enrolled in this study from children who seeked the vaccination of EV-A71 vaccine during the period from July, 2016 to June, 2018. Different sampling method were adopted in different sites. All vaccinated children were invited to participate in the study in Fengtai and Dongtai, however, systematic sampling method were adopted in other sites. Active surveillance was conducted and information about adverse reactions (ARs) occurred in 30 min, 3 d and 30 d following each dose of EV-A71 immunization was collected by field observation, phone-call or face-to-face interview. The incidence of ARs in different types, symptoms and grades were described. Results: In total, there were 45 239 children who received 71 243 doses EV-A71 vaccine. The overall incidence of ARs was 1.079% (769 doses), with the highest incidence of 1.182% (177/14 973) in 5-11 month group and the lowest incidence of 0.849% (18/2 119) in ≥ 36 month group among different age groups. There was a higher incidence in solicited ARs, which was 1.047% (746 doses). The incidences of grade 1 and grade 2 ARs were also higher, which were 0.404% (288 doses) and 0.554% (395 doses), respectively. No grade 4 ARs occurred. The doses of the first and the second vaccination was 40 736 and 30 507, respectively, and the incidences of ARs were 1.281% (522 doses) and 0.810% (247 doses). Also, the incidences of ARs were 0.091% (37 doses) and 0.043% (13 doses) in local, and 1.168% (476 doses) and 0.760% (232 doses) in system. The symptoms of ARs after the two doses of vaccination were basically the same. Redness at the injection site was the most common local ARs after each dose vaccination, with doses of 24 and 11, while fever was the most common systemic ARs, with doses of 362 and 190. Moreover, ARs mainly occurred in 30 min to 3 d after each dose vaccination, with incidence of 1.016% (414 doses) and 0.698% (213 doses) in the first and second dose, respectively. Conclusion: The ARs had a low incidence after vaccination in children and most were mild or moderate. EV-A71 vaccine with good safety is suitable for inoculation in a large scale.

[Role of endoplasmic reticulum stress-mediated glycogen synthase kinase 3ß activity in pathogenesis of acute liver failure in mice].

Objective: To study the role of endoplasmic reticulum (ER) stress-mediated glycogen synthase kinase 3ß (GSK3ß) activity in the pathological process of liver injury in acute liver failure (ALF) mice. Methods: ALF model was established by intraperitoneal injection of D-galactosamine/lipopolysaccharide (D-GalN/LPS) in C57BL/6 mice. The mice were divided into control group (n=10), ALF model group (n=18), 4-phenylbutyrate (4-PBA, an ER stress inhibitor) group (n=18) and SB216763 (a specific inhibitor of GSK3ß) group (n=16). The serum alanine transaminase (ALT) and aspartate transaminase (AST) levels were measured to reflect the liver function, liver injury was assessed by observing pathological changes of liver tissue, the levels of proteins in liver tissue were analyzed by Western blotting, the activity of GSK3ß in liver tissue was detected using GSK3ß activity assay kit, and the survival rate of hepatocyte was measured by methyl thiazolyl tetrazolium (MTT) assay. Results: In in vivo experiment, the expression levels of ER stress markers, glucose-regulated protein 78 (GRP78) and CCAAT/enhancer-binding protein homologous protein (CHOP), were upregulated during the progression of D-GalN/LPS-induced ALF, indicating activation of severe ER stress and increased activity of GSK3ß. Compared with the model group, inhibition of ER stress by 4-PBA improved liver function[ALT: (365.4±58.6) U/L vs (1 094.5±201.5) U/L, P<0.05; AST: (555.1±60.8) U/L vs (1 444.3±533.7) U/L, P<0.05)and pathological injury, also decreased the activity of GSK3ß (2.6±0.3 vs 4.6±1.3, P<0.05). Inhibition of GSK3ß activity was shown to alleviate liver injury in ALF by reducing the expression levels of GRP78 and CHOP. The in vitro experiment of tunicamycin-induced hepatocyte apoptosis showed that inhibition of GSK3ß activity increased the cell survival rate. Conclusion: In ALF induced by D-GalN/LPS, severe ER stress may accelerate the development and progress of ALF by upregulating the activity of GSK3ß.