ABSTRACT
Objective To investigate the value of cytokines in bile combined with clinical indices in predicting the degree of liver injury after liver transplantation. MethodsA total of 16 patients undergoing liver transplantation who were hospitalized in Center of Organ Transplantation, The Affiliated Hospital of Qingdao University, from January to December 2018 were enrolled, and according to the level of alanine aminotransferase (ALT) on day 1 after surgery, the patients were divided into mild liver injury (ALT <500 U/L) group with 10 patients and severe liver injury (ALT >500 U/L) group with 6 patients. Bile samples were collected on days 1, 3, 5, and 7 after surgery, and MILLIPLEX assay was used to measure the levels of 17 cytokines. R software was used to perform principal component analysis (PCA) of bile cytokines and clinical indices and gene ontology (GO) enrichment analysis of bile cytokines. The two-independent-samples t-test was used for comparison of normally distributed continuous data between two groups; The Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups. A Spearman correlation analysis was performed to evaluate the correlation between clinical indices and bile cytokines. ROC curve analysis was used to evaluate the predictive value of cytokines in bile and clinical indices for liver injury after liver transplantation. ResultsCompared with the mild liver injury group, the severe liver injury group had significantly higher expression levels of bile Fractalkine (Z=-2.828, P=0.003), soluble CD40 ligand (sCD40L) (Z=-2.850, P=0.008), interleukin-4 (Z=-2.398, P=0.017), CXCL10 (Z=-2.475, P=0.023), and macrophage inflammatory protein-1α (Z=-1844, P=0.043). The correlation analysis showed that on day 1 after liver transplantation, aspartate aminotransferase, ALT, and lactate dehydrogenase were positively correlated with the levels of several cytokines in bile (all P<0.05). The area under the ROC curve of Fractalkine, sCD40L and AST were 0.933 (0.812-1.000), 0.833 (0.589-1.000) and 0.917 (0.779-1.000), respectively, suggesting that AST and Fractalkine and sCD40L in bile on the first day after liver transplantation have significant predictive value for liver injury. The results of PCA showed that bile cytokines combined with clinical indices on day 1 after liver transplantation could better distinguish the patients with mild liver injury from those with severe liver injury. GO analysis showed that bile cytokines were associated with positive feedback regulation of external stimulus, cell chemotaxis, receptor ligand activity, cytokine activity, and cytokine-cytokine receptor interaction. ConclusionFractalkine and sCD40L in bile can predict the degree of liver injury after liver transplantation.
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Objective To observe the effects of long-term low dose arsenic exposure through drinking water on learning ability of different generations of C3H and Balb/c mice.Methods Mice (C3H and Balb/c) were exposed to arsenic at 0 mg/L (control) and 85 mg/L (20 female mice and 10 male mice per group).The control group and F1,F2,F3 and F4 mice were selected and divided into 5 experimental groups,8 mice in each group.Their offsprings were detected by the Morris water maze test (the average escape latency of 1 to 5 days) and spatial probe test (the times of through target area on the sixth day).Statistical analysis was performed with SPSS 18.0 software.Results The average escape latencies of 1 to 5 days in C3H control group were (48.09 ± 2.63),(46.09 ± 3.27),(42.72 ± 3.29),(39.31 ± 2.69) and (36.75 ± 3.92) s,F1 were (49.59 ± 3.29),(47.34 ± 3.01),(44.28 ± 6.58),(44.50 ±1.67) and (42.16 ± 2.27) s,F2 were (51.41 ± 0.78),(48.88 ± 1.45),(45.54 ± 1.46),(43.94 ± 1.69) and (42.22 ± 3.27) s,F3 were (50.91 ± 4.20),(49.78 ± 5.18),(48.03 3.45),(46.16 ± 4.42) and (44.06 ± 1.04) s,F4 were (52.66 ± 4.60),(52.38 ± 5.78),(49.06 ± 1.22),(47.69 ± 2.34) and (46.47 ± 1.56) s.The average escape latencies of Balb/c control group were (50.91 ± 2.84),(47.03 ± 4.22),(45.56 ± 4.53),(39.72 ± 5.90) and (36.22 ± 4.85) s,F1 were (50.47 ±3.20),(48.25 ± 6.53),(47.13 ± 1.25),(43.72 ± 4.27) and (40.66 ± 4.52) s,F2 were (51.31 ± 4.73),(48.88 ± 1.53),(46.56 ± 1.43),(44.25 ± 1.16) and (41.20 ± 3.79) s,F3 were (51.72 ± 3.54),(50.78 ± 4.45),(45.03 ± 3.56),(41.19 ±5.63) and (42.81 ± 6.29) s,F4 were (53.34 ± 4.60),(52.34 ± 2.77),(48.72 ± 5.92),(46.97 ± 7.38) and (44.94 ± 1.75) s.On the fourth and fifth days of F1,F2,F3 and F4 generations of C3H,the escape latencies between generations were significantly different (all P < 0.05).The times of through target area in the sixth day of the C3H control group and F1,F2,F3 and F4 mice were 2.25,1.75,1.63,1.50 and 1.38,Balb/c were 2.13,1.75,1.63,1.38 and 1.13.Conclusion Arsenic accumulation due to serial passage of C3H and Balb/c through long-term low doses arsenic exposure through drinking water has resulted in decreased learning and memory ability.
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We constructed transgenic chicken bioreactor vector, driven by chicken ovalbumin promoter, lentiviral vector and cytomegalovirus (CMV) promoter control vector encoding green fluorescent protein (GFP) and luciferase (Luc) as reporter genes. The three vectors were used to transfect or infect chicken primary oviduct epithelial cells, embryo fibroblasts cells, mouse 3T3-L1 preadipocytes cells and bovine mammary epithelial cells. High efficient and specific expression vector for transgenic chicken bioreactor was determined by detecting fluorescence and luciferase activity. Reporter gene analysis showed that chicken ovalbumin promoter expression vector was not cell type-specific in these four different cells. Additionally, luciferase reporter analysis illustrated that the chicken ovalbumin promoter activity was over 100 times lower than that of the CMV promoter in four different cells. Both of these two reporter genes were expressed in those four different cells infected by lentiviral expression vectors. Similarly, the GFP reached the similar expression level in cells infected by lentivirus and cells transfected with CMV promoter plasmid vectors when the multiplicity of infection was 20. In conclusion, the transgenic chicken bioreactor vector under the control of chicken ovalbumin promoter was not highly efficient and cell type-specific. However, the efficient expression and extensiveness oflentiviral vector could be used for studying chicken oviduct bioreactor.