The subjects of literature are the direct expression of the author’s research results. Mining valuable knowledge helps to save time for the readers to understand the content and direction of the literature quickly. Therefore, the co-occurrence network of high-frequency words in the bioinformatics literature and its structural characteristics and evolution were analysed in this paper. First, 242,891 articles from 47 top bioinformatics periodicals were chosen as the object of the study. Second, the co-occurrence relationship among high-frequency words of these articles was analysed by word segmentation and high-frequency word selection. Then, a co-occurrence network of high-frequency words in bioinformatics literature was built. Finally, the conclusions were drawn by analysing its structural characteristics and evolution. The results showed that the co-occurrence network of high-frequency words in the bioinformatics literature was a small-world network with scale-free distribution, rich-club phenomenon and disassortative matching characteristics. At the same time, the high-frequency words used by authors changed little in 2–3 years but varied greatly in four years because of the influence of the state-of-the-art technology.
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