Objective To express the anti-IL-1βscfv and soluble TNF receptor 1 (sTNFR1),and analyze their bio-activities.Methods sTNFR1 was obtained by RT-PCR from the total RNA of HeLa cells,and fused with IL-1βscfv by the hinge fragment of IgG molecule.The fusion gene IL-1scfv:TNFR1 was cloned into the expression vector pET27b(+).The fusion protein was expressed and purified from inclusion bodies.Results The ELISA analysis showed that the fusion protein could bind hIL-1β and hTNF-α respectively in a dose-dependent manner,indicating that scfv and sTNFR in the fusion protein can form the correct spatial configuration.The dolt-blot analysis showed that the fusion protein could concurrently bind with hIL-1β and hTNF-α,indicating that the combination of the two parts of the fusion protein does not influence each other for binding to their target molecules.The bioactivity assay showed that the fusion protein could inhibit both the cytotoxicity of hTNF-α on L929 cells and hIL-1β-induced proliferation of L929 cells,indicating that the fusion protein has the ability to neutralize hTNF-α and hIL-1β.Conclusion A bispecific fusion protein IL-1scfv:TNFR1 was successfully constructed.The fusion protein has the ability to inhibit the biological activity of hTNF-α and hIL-1β,and provides a drug candidate for the treatment of rheumatoid arthritis.

FGF21 (fibroblast growth factor 21) is a recently described member of the FGF family. It has been previously demonstrated that FGF21 is a potent regulator of glucose homeostasis. To improve stability of FGF21 for better efficacy, a new form of recombinant FGF21 was generated by fusion of a full length FGF21 gene and the Fc fragment of human IgG4 with flexible linker sequence. To examine the glucose regulation activity of FGF21-L-Fc, 3T3-L1 pre-adipocytes were differentiated into adipocytes, and glucose uptake activity of FGF21-L-Fc was examined by glucose oxidase and peroxidase (GOD-POD) assay. The results showed that in comparison with wild type FGF21, FGF21-L-Fc was more potent in stimulation of glucose uptake by 3T3-L1. In vivo studies on the modified protein demonstrated that FGF-L-Fc had a better efficacy in lowering blood glucose of the STZ-induced diabetic animals and controlled glucose level for a longer time. The results provided a sound basis for further studies.

Objective:To compare the differences in population distribution and prognosis of patients with nasopharyngeal carcinoma (NPC) treated with intensity-modulated radiotherapy (IMRT) in T staging of the Union for International Cancer Control (UICC) 7th edition and UICC 8th edition, and to analyze the prognostic factors in patients with NPC.Methods:The clinicopathologic date of 184 patients with newly diagnosed NPC treated with IMRT at the Department of Radiation Oncology of Weifang People′s Hospital of Shandong Province from June 1, 2005 to December 31, 2017 were retrospectively analyzed. All patients were restaged according to the 7th and 8th edition of the UICC staging system. The distribution of T staging of patients in the two staging systems was analyzed, and the consistency of the two staging systems was compared using the Kappa consistency test. Kaplan-Meier method was used for survival analysis, and log-rank test was used to compare the prognostic differences among T stages. Cox regression model was used to analyze the prognostic factors of patients with NPC.Results:Of all 184 patients with NPC, stage T 1, T 2, T 3 and T 4 respectively accounted for 18.5% (34/184), 16.8% (31/184), 15.2% (28/184) and 49.5% (91/184) according to the 7th edition UICC staging system. However, stage T 1, T 2, T 3 and T 4 respectively accounted for 18.5% (34/184), 34.2% (63/184), 30.4% (56/184) and 16.8% (31/184) according to the 8th edition UICC staging system. The T staging population distribution of the two staging systems showed moderate consistency (Kappa=0.58). There was a statistically significant difference in overall survival (OS) among patients with stage T 1, T 2, T 3, T 4 according to the 7th edition UICC staging system ( χ2=10.606, P=0.014). There were statistically significant differences in OS between stage T 1 and stage T 2, T 3, T 4 ( χ2=4.866, P=0.027; χ2=11.965, P=0.001; χ2=4.351, P=0.037). The OS curves of stage T 2 and T 4 could not be separated. Moreover, the OS curves of stage T 3 and T 4 were distributed in reverse order. There was a statistically significant difference in OS among patients with stage T 1, T 2, T 3, T 4 according to the 8th edition staging system ( χ2=8.663, P=0.034). There were statistically significant differences in OS between stage T 1 and stage T 3, T 4( χ2=8.746, P=0.003; χ2=7.580, P=0.006). The OS curves of stage T 1 to T 4 were distributed in order, but the curves of stage T 3 and T 4 could not be separated. There was a statistically significant difference in progression-free survival (PFS) among patients with stage T 1, T 2, T 3, T 4 according to the 7th edition UICC staging system ( χ2=11.289, P=0.010). There were statistically significant differences in PFS between stage T 1 and stage T 2, T 3, T 4 ( χ2=8.209, P=0.004; χ2=13.302, P<0.001; χ2=6.550, P=0.010). The PFS curves of stage T 2 and T 4 could not be separated. Moreover, the PFS curves of stage T 3 and T 4 were distributed in reverse order. There was a statistically significant difference in PFS among patients with stage T 1, T 2, T 3, T 4 according to the 8th edition staging system ( χ2=12.074, P=0.007). There were statistically significant differences in PFS between stage T 1 and stage T 2, T 3, T 4( χ2=5.182, P=0.023; χ2=11.217, P=0.001; χ2=10.174, P=0.001). The PFS curves of stage T 1 to T 4 were distributed in order, but the curves of stage T 3 and T 4 could not be separated. The results of Cox multivariate analysis showed that T staging of both staging systems were the independent prognostic factors of the OS ( P=0.013; P=0.026) and PFS ( P=0.031; P=0.012). However, T staging of the two editions were not the independent prognostic factors of the local recurrence-free survival (LRFS) ( P=0.351; P=0.167) and distant metastasis-free survival (DMFS) ( P=0.059; P=0.052). The age was the independent prognostic factor of the OS ( HR=2.70, 95% CI: 1.53-4.76, P=0.001; HR=2.74, 95% CI: 1.55-4.84, P=0.001), PFS ( HR=2.72, 95% CI: 1.46-5.08, P=0.002; HR=2.94, 95% CI: 1.57-5.52, P=0.001), LRFS ( HR=5.87, 95% CI: 1.62-21.27, P=0.007; HR=6.02, 95% CI: 1.61-22.49, P=0.008) and DMFS ( HR=2.40, 95% CI: 1.22-4.72, P=0.011; HR=2.63, 95% CI: 1.34-5.18, P=0.005). N staging was the independent prognostic factor of the OS ( P=0.031; P=0.028). Conclusion:The T staging population distribution of the 7th and 8th edition UICC staging system had moderate consistency, and the T staging of the 8th edition is more advantageous in predicting the prognosis of OS and PFS. In both editions, T staging is an independent prognostic factor for OS and PFS.