Salient segmentation is a critical step in biomedical image analysis, aiming to cut out regions that are most interesting to humans. Recently, supervised methods have achieved promising results in biomedical areas, but they depend on annotated training data sets, which requires labor and proficiency in related background knowledge. In contrast, unsupervised learning makes data-driven decisions by obtaining insights directly from the data themselves. In this paper, we propose a completely unsupervised self-aware network based on pre-training and attentional backpropagation for biomedical salient segmentation, named as PUB-SalNet. Firstly, we aggregate a new biomedical data set from several simulated Cellular Electron Cryo-Tomography (CECT) data sets featuring rich salient objects, different SNR settings, and various resolutions, which is called SalSeg-CECT. Based on the SalSeg-CECT data set, we then pre-train a model specially designed for biomedical tasks as a backbone module to initialize network parameters. Next, we present a U-SalNet network to learn to selectively attend to salient objects. It includes two types of attention modules to facilitate learning saliency through global contrast and local similarity. Lastly, we jointly refine the salient regions together with feature representations from U-SalNet, with the parameters updated by self-aware attentional backpropagation. We apply PUB-SalNet for analysis of 2D simulated and real images and achieve state-of-the-art performance on simulated biomedical data sets. Furthermore, our proposed PUB-SalNet can be easily extended to 3D images. The experimental results on the 2d and 3d data sets also demonstrate the generalization ability and robustness of our method.