ObjectiveTo investigate the functional role and underlying molecular mechanisms of fumarylacetoacetate hydrolase （FAH） in the progression of glioblastoma （GBM）. MethodsDifferential expression analysis was performed on the TCGA-GBM， GSE4290， and GSE116520 datasets. Weighted gene co-expression network analysis （WGCNA） was used to identify key modules， and Cox regression and risk modeling were used to screen prognostic genes. Immune infiltration analysis of prognostic genes was carried out by using single-cell RNA sequencing panels. The clinical expression signature of FAH in GBM was analyzed in the TCGA and HPA databases. The functional role of FAH was validated by in vitro and in vivo experiments， and pathway analysis was performed to explore the underlying mechanisms. ResultsA total of 152 overlapping genes were identified across the three GBM datasets （P<0.05）. WGCNA revealed that the turquoise module was most strongly associated with tumor purity， stromal score， immune score， and ESTIMATE score （P<0.001）. Compared with normal tissues， three prognostic genes （CTSD， FAH， and THBD） were upregulated in GBM and correlated with immune infiltration （P<0.05）. FAH mRNA and protein levels were elevated in GBM tissues relative to normal tissues， and its expression was significantly associated with age stratification and TP53 mutation （P<0.05）. CCK-8 assay results showed that， compared with the shNC group， the proliferative activity of GBM cells in the shFAH group was reduced （P<0.001）. Transwell migration and invasion assays demonstrated that， relative to the shNC group， the numbers of migrated and invaded cells in the shFAH group decreased （P<0.05）. Western blot analysis revealed that the protein expression levels of PI3K， p-AKT， and p-mTOR in the shFAH group decreased compared with those in the shNC group （P<0.05）. In vivo subcutaneous xenograft experiments further confirmed that tumor volume and weight significantly decreased in the shFAH group compared with the shNC group （P<0.001）. ConclusionFAH promotes GBM progression by activating the PI3K/AKT/mTOR signaling pathway and may serve as a potential therapeutic target for GBM.

BACKGROUND: Arsenic trioxide (ATO) is indicated as a broad-spectrum medicine for a variety of diseases, including cancer and cardiac disease. While the role of ATO in hepatic ischemia/reperfusion injury (HIRI) has not been reported. Thus, the purpose of this study was to identify the effects of ATO on HIRI. METHODS: In the present study, we established a 70% hepatic warm I/R injury and partial hepatectomy (30% resection) animal models in vivo and hepatocytes anoxia/reoxygenation (A/R) models in vitro with ATO pretreatment and further assessed liver function by histopathologic changes, enzyme-linked immunosorbent assay, cell counting kit-8, and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay. Small interfering RNA (siRNA) for extracellular signal-regulated kinase (ERK) 1/2 was transfected to evaluate the role of ERK1/2 pathway during HIRI, followed by ATO pretreatment. The dynamic process of autophagic flux and numbers of autophagosomes were detected by green fluorescent protein-monomeric red fluorescent protein-LC3 (GFP-mRFP-LC3) staining and transmission electron microscopy. RESULTS: A low dose of ATO (0.75 μmol/L in vitro and 1 mg/kg in vivo ) significantly reduced tissue necrosis, inflammatory infiltration, and hepatocyte apoptosis during the process of hepatic I/R. Meanwhile, ATO obviously promoted the ability of cell proliferation and liver regeneration. Mechanistically, in vitro  studies have shown that nontoxic concentrations of ATO can activate both ERK and phosphoinositide 3-kinase-serine/threonine kinase (PI3K-AKT) pathways and further induce autophagy. The hepatoprotective mechanism of ATO, at least in part, relies on the effects of ATO on the activation of autophagy, which is ERK-dependent. CONCLUSION Low, non-toxic doses of ATO can activate ERK/PI3K-AKT pathways and induce ERK-dependent autophagy in hepatocytes, protecting liver against I/R injury and accelerating hepatocyte regeneration after partial hepatectomy.
Animals ; Arsenic Trioxide ; Autophagy/physiology* ; Reperfusion Injury/prevention & control* ; Mice ; Male ; Proto-Oncogene Proteins c-akt/physiology* ; Arsenicals/therapeutic use* ; Oxides/therapeutic use* ; Liver/metabolism* ; Extracellular Signal-Regulated MAP Kinases/metabolism* ; Mice, Inbred C57BL

Purpose: Although obesity is associated with numerous diseases, the risks of disease may depend on metabolically healthy status. Nevertheless, it is unclear to whether metabolically healthy status affects risk of gastrointestinal (GI) cancer in general Chinese population. Materials and Methods: A total of 114,995 participants who met the criteria were included from the Kailuan Study. The study participants were divided into four groups according to body mass index (BMI)/waist circumference (WC) and metabolic status. Incident of GI cancer (esophageal cancer, gastric cancer, liver cancer, biliary cancer, pancreatic cancer, and colorectal cancer) during 2006-2020 were confirmed by review of medical records. The Cox proportional hazard regression models were used to assess the association metabolically healthy status with the risk of GI cancer by calculating the hazard ratios (HR) and 95% confidence interval (CI). Results: During a mean 13.76 years of follow-up, we documented 2,311 GI cancers. Multivariate Cox regression analysis showed that compared with the metabolically healthy normal-weight group, metabolically healthy obese (MHO) participants demonstrated an increased risk of developing GI cancer (HR, 1.54; 95% CI, 1.11 to 2.13) by BMI categories. However, such associations were not found for WC category. These associations were moderated by age, sex, and anatomical site of the tumor. Individuals with metabolic unhealthy normal-weight or metabolic unhealthy obesity phenotype also have an increased risk of GI cancer. Conclusion MHO phenotype was associated with increased risk of GI cancer. Moreover, individuals who complicated by metabolic unhealthy status have an increased risk of developing GI cancer. Hence, clinicians should consider the risk of incident GI cancer in people with abnormal metabolically healthy status and counsel them about metabolic fitness and weight control.

T cells, including both CD4⁺ and CD8⁺ T cells, play a pivotal role in mediating various inflammation and immune disorders. A long-standing challenge in T cell-based immunotherapy is to precisely inactivate or delete the pathogenic T cells in inflammation and autoimmune diseases, or to selectively expand the immunocompetent T cell in tumor or other immune compromised situations, without inducing global immunosuppression or zealous immune activation respectively. To achieve this, a specific marker is needed to differentiate the pathogenic or immunocompetent T cell among the rests. Indeed, recent progress of immunology strongly suggests that CXC chemokine receptor 6 (CXCR6, CD186) is such a kind of marker. Here, we review the emerging role of CXCR6 as a novel target for immunotherapy and discuss the underlying mechanism. We propose that CXCR6-based immunotherapy will play a significant role in autoimmune, nonalcoholic steatohepatitis (NASH), tumor, coronavirus disease 2019 (COVID-19) and even ageing-related inflammatory infliction.

Objective:To observe any effect of electroacupuncture (EA) on the expression of phosphorylated extracellular signal-regulated protein kinase (p-ERK1/2) and phosphorylated cyclic adenosine monophosphate response element binding protein (p-CREB) in the spinal dorsal horns of diabetics experiencing neuropathic pain.Methods:Eight rats were randomly selected from 30 healthy male Sprague-Dawley rats as the normal group (N), and the remaining twenty-two rats were treated with a single high-dose intraperitoneal injection of streptozotocin (STZ) to establish a neuropathic pain model. The rats modeled successfully were randomly divided into a model group (M, n=8) and an EA group ( n=8). In the EA group, electroacupuncture was applied at the bilateral Hou san li and Kunlun acupoints starting on the 15th day after the STZ injection. The daily sessions lasted 30 minutes for 1 week. Body weight (BW), fasting blood glucose (FBG) and paw withdrawal latency (PWL) were observed before the STZ injection and on the 7th, 14th, and 21st days afterward. The expression of p-ERK1/2 and p-CREB in the dorsal horns of the rats′ spinal cords was detected using western blotting. The count of p-CREB-positive cells in the dorsal horns and their co-localization with neurons was detected using immunofluorescence. Results:In comparison with the N group, the average BW of the M group on the 7th, 14th and 21st days after the STZ injection was significantly lower, while the average FBG was significantly higher. There was no significant difference between the M and N groups in the average PWL on the 7th day after the STZ injection, but it had decreased significantly in the M group on the 14th and 21st days. Compared with the M group, the average PWL of the EA group was significantly longer on the 21st day after the injection. The expression of p-ERK1/2 and p-CREB protein in the spines of the M group was significantly higher than in the N group. p-CREB positive cells were more numerous in the M group compared with the N group, while in the EA group they were fewer. P-CREB was co-located with neurons in the spinal dorsal horn.Conclusion:EA can alleviate neuropathic pain effectively, perhaps by inhibiting the expression of p-ERK1/2 and p-CREB in the dorsal horns of the spinal cord.

[Abstract] Objective：To investigate the expression of ITGB2 (integrinβ2) in renal cell carcinoma (RCC) tissues and cells (ACHN cells) and its effects on the proliferation, migration, invasion and cell cycle of ACHN cells. Methods: The expression level of ITGB2 in RCC tissues and para-cancerous tissues was analyzed by GEPIA database. The tissue samples of 66 RCC patients retained in the biological specimen bank of the Fourth Hospital of Hebei Medical University from 2016 to 2020 were selected for this study. The expression level of ITGB2 in 66 cases of RCC tissues and para-cancerous tissues was detected by immunohistochemical SP and qPCR, and the relationship between ITGB2 and clinical parameters was analyzed. The shRNA with ITGB2 knockdown was constructed and transfected into ACHN cells for functional experiments to detect its effect on the malignant biological behaviors of ACHN cells, and its effect on the cell cycle was detected by flow cytometry. WB was used to detect the effect of ITGB2 knockdown on ITGB2 protein expression in ACHN cells. Results: The relative expression of ITGB2 in RCC tissues was significantly higher than that in para-cancerous tissue (P<0.01), and the expression was related to the clinical stage of RCC (P<0.05). Transfection of shITGB2 into ACHN cells could knock down the gene and protein expression of ITGB2 (all P<0.01). Knockdown of ITGB2 could significantly inhibit the proliferation (P<0.05), migration(P<0.01) and invasion (P<0.05) of ACHN cells but had no significant effect on cell cycle (P>0.05). Conclusion: ITGB2 is highly expressed in RCC tissues and cells and is associated with the clinical stage of RCC. Knockdown of ITGB2 can inhibit the malignant biological behaviors of ACHN cells.

Objective:To explore the relationship between the changes of total cholesterol (TC), C-reactive protein (CRP), vascular endothelial growth factor (VEGF) and the degree of false lumen thrombosis after thoracic endovascular aortic repair (TEVAR) and its clinical significance.Methods:A total of 95 patients with aortic dissection admitted to the Affiliated Hospital of Jining Medical College from June 2015 to July 2020 were selected for retrospective study. All patients were treated with TEVAR. According to the disappearance of false lumen detected by computed tomography (CT) angiography six months after operation, 95 patients were divided into complete disappearance group ( n=43) and incomplete disappearance group ( n=52). The levels of plasma TC, CRP and VEGF in the two groups were compared before operation and 1 and 3 months after operation, as well as the degree of false lumen thrombosis. Spearman′s method was used to analyze the relationship between the levels of plasma TC, CRP and VEGF and the degree of postoperative false lumen thrombosis; multivariate logistic regression was used to analyze the factors affecting the disappearance of false lumen after TEVAR; The receiver operating characteristic (ROC) curve and area under the curve (AUC) were used to analyze the value of each index in predicting the complete disappearance of false lumen. Results:The plasma levels of TC, CRP and VEGF in the complete disappearance group were lower than those in the incomplete disappearance group 1 and 3 months after operation (all P<0.05). The degree of false lumen thrombosis in the complete disappearance group was significantly different from that in the incomplete disappearance group 1 and 3 months after operation (all P<0.05). The plasma levels of TC, CRP and VEGF 1 and 3 months after TEVAR were negatively correlated with the degree of false lumen thrombosis (all P<0.05). Multivariate logistic regression analysis showed that the plasma levels of TC, CRP and VEGF 1 and 3 months after operation were correlated with the disappearance of false lumen (all P<0.05). With the passage of time, the AUC of each index to predict the complete disappearance of false lumen gradually increased. At 3 months after operation, the AUC of TC, CRP, VEGF and combined prediction of the complete disappearance of false lumen were 0.706, 0.899, 0.781 and 0.943, respectively (all P<0.05). Conclusions:The changes of plasma TC, CRP and VEGF levels after TEVAR are related to the degree of false lumen thrombosis and the disappearance of false lumen in patients with aortic dissection. Combined examination of the three can be an effective method to predict the complete disappearance of false lumen.

Objective:To investigate the effects of TNF-α knockout on liver and spleen neutrophil responses to Vibrio vulnificus bloodstream infection in a mouse model. Methods:(1) TNF-α-knockout (TNF-α -/-) and wild-type (WT) C57BL/6J mice aged 6-8 weeks were randomly divided into four groups with six in each group: uninfected WT group, infected WT group, uninfected TNF-α -/- group and infected TNF-α -/- group. The mouse model of bloodstream infection was constructed by intraperitoneal injection of Vibrio vulnificus CGMCC1.1758 (2×10 8 CFU/200 μl), while the mice in the uninfected groups were injected intraperitoneally with equal amount of PBS. (2) Liver immune cells and splenocytes were isolated 4 h after infection and subjected to analyze the percentages and numbers of neutrophils, and the changes in cell viability, cellular reactive oxygen species (ROS) level and phagocytosis by flow cytometry. In addition, effects of Vibrio vulnificus bloodstream infection on mTOR signaling pathway in murine neutrophils were evaluated in vivo. Results:(1)Compared with the uninfected WT group, the percentages and numbers of neutrophils in liver and spleen tissues of the infected WT group increased significantly. The percentage and number of liver neutrophils were significantly higher in the infected TNF-α -/- group than in the infected WT group, but no significant difference in spleen neutrophils was detected between the two groups. (2) Compared with the infected WT group, the phagocytosis of liver neutrophils rather than that of spleen neutrophils was enhanced in the infected TNF-α -/- group. (3) The survival rates of neutrophils in both liver and spleen were decreased, while the cellular ROS level was significantly increased in the infected WT group compared with those of the uninfected WT group. Compared with the infected WT group, the infected TNF-α -/- group had increased survival rates of both liver and spleen neutrophils, but decreased level of ROS. (4) The levels of p-AKT (S473) in liver and spleen neutrophils of the infected WT group were lower than those of the uninfected WT group. Compared with the infected WT group, the infected TNF-α -/- group had lower level of p-AKT (S473) in liver neutrophils, but higher p-AKT (S473) level in spleen neutrophils. There were no significant differences in p-4E-BP1(T37/46) levels between the uninfected WT group and the infected WT group. The p-4E-BP1 (T37/46) level in liver neutrophils was lower in the infected TNF-α -/- group than in the infected WT group, but no significant difference in p-4E-BP1 (T37/46) levels in spleen neutrophils was observed between the two groups. Conclusions:TNF-α had different effects on the neutrophils in spleen and liver tissues of mice with Vibrio vulnificus bloodstream infection. It played a critical role in regulating the recruitment, phagocytic function and mTOR signaling of liver neutrophils after Vibrio vulnificus infection in vivo.

Objective To investigate the feasibility,safety and efficacy of endovascular recanalization of the symptomatic occlusion of large intracranial artery in anterior circulation.Methods From October 2015 to December 2017,13 patients with symptomatic non-acute occlusion of large intracranial artery in anterior circulation were enrolled into this study and underwent endovascular recanalization.The initial procedural results,including the rate of successful recanalization and perioprocedural complications,and angiographic and clinical follow-up results were collected.The functional outcome was evaluated at discharge and 90 days.Results Recanalization was successful in 11 out of 13 patients.Perioperative complications occurred in 8 cases,including distal embolization in 7 cases (3 with symptom and 4 without),in which intracerebral hemorrhage associated with embolectomy was found in 1 case;and distal embolization concomitant with artery dissection in 1 case.At discharge,the symptoms of 10 out of 11 patients with successful recanalization were improved and 1 was unchanged;one of 2 patients with recanalization failure was aggravated and 1 was unchanged.After the procedure,1 patient with successful recanalization,but complicated with intracerebral hemorrhage associated with embolectomy was lost at follow-up,thus angiographic follow-up was available in the remaining 10 patients.Of the 10 patients,1 patient developed in-stent restenosis at 12 months and 9 patients had no hemodynamic stenosis/reocclusion.The clinical follow-up was available in 12 patients.No recurrence of TIA or stroke was found in 9 cases with successful recanalization except for 1 case who developed in-stent stenosis and suffered from TIA.At the follow-up of 90 days,l0 patients with successful recanalization showed good function (mRS∶0-2),2 patients with recanalization failure were deteriorated.Conclusions In strictly selected patients with symptomatic non-acute occlusion of large intracranial artery in anterior circulation,endovascular recanalization was feasible and safe,which may improve patients' symptoms in a short term and reduce the recurrence rate of stroke,but its definite efficacy needs to be confirmed by studies with larger sample and longer follow-up.

Objective To explore the relationship between Toll-like receptor 2(TLR2) and autophagy marker protein in glioma. Methods Glioma tumors of a total of 74 patients from June 2012 to December 2017 were surgically resected, including WHO gradeⅠtoⅡ32 cases, grade Ⅲ 20 cases, gradeⅣ22 cases. Immunohistochemistry and Western blot were used to detect the expression of TLR2, autophagy related protein LC3B, Beclin 1 and apoptosis related protein Bax and Bcl-2. The correlation between TLR2 and autophagy related protein LC3B and Beclin 1 were analyzed. Results In high grade glioma (HGG) tissue and low grade glioma (LGG) tissue, the TLR2 positive expression rates were 92.9％(39/42) and 75.0％(24/32), and there was significant difference (P<0.05). In HGG tissue, autophagy related protein LC3B, Beclin1 protein was strongly positive and the positive expression rates were 45.2％(19/42) and 52.4％(22/42). In LGG tissue, LC3B and Beclin1 protein positive expression rates were 18.8％(6/32) and 15.6％(5/32), and there were significant differences (P<0.05). Spearman correlation analysis showed that TLR2 protein was closely related to autophagy related protein LC3B (r=0.5638, P<0.05) and Beclin1 (r=0.6101, P<0.05). Conclusions TLR2 is highly expressed in HGG tissue, and its expression level may be related to autophagy, which has potential value as a targeted therapy.