Analysis of the in vitro function and internalization ability of a humanized EGFR antibody AE01 expressed by Chinese hamster ovary cells.

The primary objective of this study was to obtain humanized EGFR antibody and to study it in vitro binding and endocytosis to A431 epidermoid carcinoma cells overexpressing EGFR. Firstly, humanized anti-EGFR AE01 was stably expressed in CHO system. The expression of AE01 was detected by SDS-PAGE and Western blot. The binding and endocytosis of AE01 were detected by flow cytometry and immunofluorescence assay. The results showed that: (1) Pure humanized AE01 was prepared, (2) AE01 specifically binds to A431 cells on the cell surface (EGFR-positive), but not binds to NIH 3T3 cells (EGFR-negative), (3) AE01 can effectively inhibit the proliferation of A431 cells, and (4) AE01 binds to A431 cell surface triggered internalization. The antibody is expected to be a candidate molecule for EGFR overexpressed cancer cell targeted therapeutic vectors.

The Level of HbA1c Evaluates the Extent of Coronary Atherosclerosis Lesions and the Prognosis in Diabetes with Acute Coronary Syndrome.

Background: The level of HbA1c can reflect the average level of blood glucose over 3 months, which is the gold standard indicator for monitoring blood glucose. The relationship between the level of HbA1c and the extent of coronary atherosclerosis lesions or the prognosis in diabetes with acute coronary syndrome (ACS) remains poorly understood. Aims: To explore whether the level of HbA1c can evaluate the extent of coronary atherosclerosis lesions or the prognosis in diabetes with acute coronary syndrome (ACS) using the SYNTAX score, the Global Registry of Acute Coronary Events (GRACE) score, left ventricular function (LVEF), left ventricular end-diastolic volume (LVEDV), and major adverse cardiac events (MACEs) in the hospital and 12 months after discharge. Methods: This study was a prospective, randomized, open-label, and parallel group study. Patients with diabetes with ACS were recruited into this study indiscriminately, and all the participants were divided into two groups according to the level of HbA1c: HbA1c level ≤ 7%group and HbA1c level > 7%group. The followings were used as the evaluation indicators: SYNTAX score, GRACE score, LVEF, LVEDV, and MACEs in hospital and 12 months after discharge. Results: A total of 233 patients with diabetes and ACS were enrolled and assigned to two groups according to their level of HbA1c: the HbA1c ≤ 7%group (n = 92) and the HbA1c > 7%group (n = 141). The results showed that the proportion of STEMI was higher in the HbA1c ≤7% group (p < 0.05), while the proportion of NSTEMI has not significantly higher in the HbA1c >7% group (p > 0.05). Regression analysis indicated that HbA1c level was significantly positively correlated with GRACE score (r = 0.156, F = 5.784, p = 0.017, n = 233) and SYNTAX score (r = 0.237, F = 13.788, p < 0.001, n = 233), and there were no statistically significant differences in LVEDV and LVEF between the two groups (p > 0.05). The total MACEs rate showed no significant difference between the two groups during hospitalization (p > 0.05) but showed significant differences at 12 months after discharge (p < 0.05). Conclusions: This study shows that HbA1c level was positively correlated with the extent of coronary atherosclerosis lesions and the prognosis in diabetes with ACS. The higher the HbA1c level is, the more severe the coronary atherosclerotic lesion and the worse the prognosis in diabetes with ACS are.

Effects of mercury on the structure and activity of BLM642-1290 recombinant helicase.

OBJECTIVE: Bloom's syndrome is an autosomal recessive disorder characterized by genomic instability and a predisposition to many cancers. Mutations of the BLM gene (encoding a BLM helicase) may form a structure of the etiology of this disease. As a global pollutant, mercury poses a major threat to human health. The current study was conducted to elucidate the effects of Hg(2+) on the structure and activity of BLM642-1290 recombinant helicase, and to further explore the molecular mechanisms of mercury toxicity to the DNA helicase. METHODS: The effects of Hg(2+) on biological activity and structure of BLM642-1290 recombinant helicase were determined by fluorescence polarized, ultraviolet spectroscopic, and free-phosphorus assay technologies, respectively. RESULTS: The helicase activity, the DNA-binding activity, and the ATPase activity of BLM642-1290 recombinant helicase were inhibited by Hg(2+) treatment. The LMCT (ligand-to-metal charge transition) peaks of the helicase were enhanced with the increase of the Hg(2+) level. The LMCT peaks of the same concentration of helicase gradually increased over time. CONCLUSION: The biological activity of BLM642-1290 recombinant helicase is inhibited by Hg(2+) treatment. The conformation of the helicase is significantly altered by Hg(2+). There exist two binding sites between Hg(2+) and the helicase, which are located in the amino acid residues 1063-1066 and 940-944 of the helicase, respectively.