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ObjectiveTo explore the diagnostic value of serum YKL-40 for liver fibrosis stage, and to provide a reference for noninvasive diagnosis of liver fibrosis in patients with chronic liver disease. MethodsWe searched PubMed, EMBASE, the Cochrane Library, Web of Science, and CNKI for studies on the clinical value of YKL-40 in the diagnosis of liver fibrosis or cirrhosis. The quality of studies was evaluated using the QUADAS-2 tool to assess the risk of bias. Comprehensive quantitative evaluation of the included studies was performed using Stata 12.0. The source of heterogeneity was analyzed, and the forest plot and summary receiver operating curve (SROC) were generated. ResultsA total of nine studies involving 1592 patients were included in the meta-analysis; six studies were conducted on significant fibrosis (≥F2), and seven studies were conducted on progressive fibrosis (≥F3). In the diagnosis of significant fibrosis (≥F2), YKL-40 had pooled sensitivity, specificity, positive likelihood ratio, negative likelihood ratio, odds ratio, and area under the SROC (AUC) of 078 (95% confidence interval ［CI］: 0.69-0.85), 0.53 (95%CI: 0.33-0.72), 1.7 (95%CI: 1.0-2.7), 0.41 (95%CI: 0.21-0.76), 4 (95%CI: 1-13), and 0.76 (95%CI:0.72-0.80), respectively. In the diagnosis of progressive fibrosis (≥F3), YKL-40 had pooled sensitivity, specificity, positive likelihood ratio, negative likelihood ratio, odds ratio, and AUC of 0.83 (95%CI: 0.76-089), 0.72 (95%CI: 0.62-0.80), 3.0 (95%CI: 2.0-4.4), 0.23 (95%CI: 0.14-0.37), 13 (95%CI: 5-30), and 0.85 (95%CI: 082-0.88), respectively. ConclusionThe diagnostic value of serum YKL-40 for significant and progressive liver fibrosis is limited, so it may not be a new, effective serum marker for the staging of liver fibrosis.
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To express Pleurocidin in Escherichia coli and to enhance the secretory efficiency of the fusion protein, the gene encoding Pleurocidin was ligated with Cherry DNA sequence via blunt-end ligation. Then this fusion gene was cloned into pET22b (+) vector and the recombinant plasmid was transformed into E. coli BL21 (DE3). Lactose was used to induce expression of fusion protein. The recombinant plasmid pET22b (+) -CP was successfully constructed and high-level expression of fusion protein was induced with lactose. Statistics showed that addition of glycine after 16 h of induction significantly enhanced the secretory efficiency of the fusion protein. After hydrolysis of the fusion protein by diluted hydrochloric acid and some further purification steps, r-Pleurocidin was obtained with antibacterial activity against E. coli DH5α and Bacillus subtilis BS168. In conclusion, the fusion protein was expressed in E. coli and biologically active r-Pleurocidin was obtained after hydrochloric acid cleavage and purification.
Subject(s)
Animals , Cloning, Molecular , Escherichia coli , Metabolism , Fish Proteins , Flounder , Recombinant Fusion ProteinsABSTRACT
Objective:To observe inhibitory effrects of DNA vaccine co-expressing CEA tandem repeat epitopes and FL on cancer cells in mice.Methods:The encoding sequences for CEA tandem repeats and FL were inserted into plasmid pcDNA3.0 using gene recombinant technique.BALB/c mice were immunized intramuscularly with the co-expressing DNA vaccine.The survival time and tumor size were measured and specific CTL cytotoxicity was detected by ~(125)I-UdR release method.Results:Compared with that of the control,the survival time was prolonged (P＜0.01)and the tumors were significantly inhibited in the mice immunized with the vaccine peDNA-triCEA_(625-667)-sFL(P＜0.01).The splenic cells from mice immunized with the vaccine pcDNA-triCEA_(625-667)-sFL induced strongly cytotoxicity against tumor cells H22-CEA ~+(P＜0.01).Conclusion:The recombinant DNA vaccine co-expressing pcDNA-triCEA_(625-667)-sFL can suppress the growth of tumor expressing CEA in mice and enhance CTL response against CEA antigen.
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Objective To probe the relationship between the expression of TL1A and the level of IFN-γ secreted by T cells in the acute stage of Guillain-Barre syndrome (GBS). Methods ① Six-week female Bal b/c mice were immunized by purified recombinant human soluble TNF-like molecular 1A (rhsTL1A) protein. The polyclonal antibody against rhsTL1A was identified by immunofluorescence using human umbilical vein epithelial cells (HUVEC). ② To detect the biologic activity of rhsTL1A, the peripheral blood mononuclear cells (PBMC) from the healthy donors were separated by Ficoll gradient centrifugation and were seeded on 96-well plates with medium containing 2 μg/ml PHA (control group), 2 μg/ml PHA + 25 ng/ml rhsTL1 A, 2 μg/ml PHA + 100 ng/ml rhsTL1A and 2 μg/ml PHA + 400 ng/ml rhsTLlA respectively. T cell proliferation assay was carried out using ~3H-TdR. ③ IFN-γ productions in the sera of the children with GBS in the acute stage were detected by ELISA. ④ The ratio of CD_3~+ TL1A~+ T cells to CD_3~+ T cells in the peripheral blood of the children with GBS in acute stage was detected with flow, cytometry. ⑤PBMC from the children in acute GBS were separated and cultured in the environment adding 2 μg/ml PHA and 400 ng/ml rhsTL1A in vitro. Then, the IFN-γ in the supernatant was determined by ELISA kit after 72 hours. Results ① hTL1A A expressed by eukaryotic HUVECs was recognized by rhsTL1 A polyclonal antiserum. ② The result of T cell proliferation assay showed that SI of 25 ng/ml rhTL1A, 100 ng/ml rhTL1A A and 400 ng/ml rhTL1A group was increased compared with control group. The SI of 2 μg/ml PHA +400 ng/ml rhsTL1 A group was the highest (2. 65) among them. ③ IFN-γ productions in the sera of the children with GBS in the acute stage ((102. 25±22. 17) pg/ml) were increased significantly compared with healthy control ((28.75 ± 1.31) pg/ml, t = 3. 309, P < 0. 05). ④ The ratio of CD_3~+ TL1A~+ T cells to CD_3~+ T cells in the peripheral blood of the children with GBS in acute stage (18.22%± 1.83%) was enhanced significantly compared with healthy control (5. 17% ±0. 48%, t = 6. 884, P < 0. 01). ⑤ PBMC both in healthy control and the acute GBS secreted more IFN-γ markedly ((43.56± 4.41) pg/ml and (180.64 ± 38.39) pg/ml) after being incubated in 2 μg/ml PHA and 400 ng/ml rhsTL1A (t =4. 523 and 2. 600, P <0. 01 and 0. 05 respectively). Moreover, PBMC in acute GBS secreted more IFN-γ, than that of the healthy group markedly (t = 3. 545, P < 0. 05). Conclusions ① The mouse antiserum recognizing rhsTL1A is successfully obtained. ② In this study, 400 ng/ml rhsTL1A promotes the proliferation of T cells activated by 2 μg/ml PHA, indicating that rhsTL1A has biological activity. ③ The expression of hTL1A of activated T cells in the peripheral blood of the children with acute GBS is up-regulated. These TL1A proteins promote the secretion of IFN-γ through binding to their receptors DR_3.