Traditional development of small protein scaffolds has relied on display technologies and mutation-based engineering, which limit sequence and functional diversity, thereby constraining their therapeutic and application potential. Protein design tools have significantly advanced the creation of novel protein sequences, structures, and functions. However, further improvements in design strategies are still needed to more efficiently optimize the functional performance of protein-based drugs and enhance their druggability. Here, we extended an evolution-based design protocol to create a novel minibinder, BindHer, against the human epidermal growth factor receptor 2 (HER2). It not only exhibits super stability and binding selectivity but also demonstrates remarkable properties in tissue specificity. Radiolabeling experiments with ^99mTc, ⁶⁸Ga, and ¹⁸F revealed that BindHer efficiently targets tumors in HER2-positive breast cancer mouse models, with minimal nonspecific liver absorption, outperforming scaffolds designed through traditional engineering. These findings highlight a new rational approach to automated protein design, offering significant potential for large-scale applications in therapeutic mini-protein development.

Objective:To investigate the clinical efficacy of five-element music therapy in the treatment of bipolar disorder.Methods:Eighty patients with bipolar disorder admitted to Shandong Mental Health Center from January 2018 to December 2019 were included in this study. They were randomly divided into a study group and a control group ( n = 40/group). The control group was treated with conventional drug treatment. The study group was treated with five-element music therapy based on conventional drug treatment. Before, during and after treatment, mania and depression were evaluated. Symptom self-evaluation scale and the Traditional Chinese Medicine Five State Emotion Questionnaire were completed to evaluate the curative effect. Patients were followed up by telephone call. Recurrence at 3, 6 and 12 months was recorded. Results:Before treatment, there were no significant differences in the scores of the Bech-Rafaelsen Mania Rating Scale (BRMs), Hamilton Depression Rating Scale (HAMD), Traditional Chinese Medicine Five State Emotion Questionnaire, Symptom Checklist-90 (SCL-90) between the two groups (all P > 0.05). During and after treatment, BRMs, HAMD, Traditional Chinese Medicine Five State Emotion Questionnaire, and SCL-90 scores were significantly decreased in each group compared with before treatment [Control group: BRMs score during and after treatment: (11.52 ± 1.57) points, (9.43 ± 1.05) points; HAMD score during and after treatment: (16.75 ± 3.05) points, (13.61 ± 2.51) points; Traditional Chinese Medicine Five State Emotion Questionnaire: emotional vocabulary (67.25 ± 5.7) points, (56.38 ± 4.97) points, physical symptoms (29.52 ± 3.94) points, (24.19 ± 3.05) points, SCL-90 score: (100.52 ± 10.26) points, (68.85 ± 7.33) points. Study group: BRMs score during and after treatment: (9.33 ± 1.09) points, (7.85 ± 0.82) points; HAMD score: (13.74 ± 2.54) points, (10.17 ± 1.97) points; Traditional Chinese Medicine Five State Emotion Questionnaire: emotional vocabulary: (58.19 ± 5.06) points, (46.85 ± 4.06) points, physical symptoms (25.14 ± 3.54) points, (20.11 ± 2.57) points; SCL-90 scores: (90.85 ± 8.97) points, (56.87 ± 5.81) points]. During and after treatment, scores of BRMs, HAMD, Traditional Chinese Medicine Five State Emotion Questionnaire and SCL-90 in the study group were significantly lower than those in the control group ( t = 3.77, 7.01, 4.48, 6.99, 8.78, 4.89, 6.05, 4.19, 7.57, all P < 0.05). Total response rate in the study group was significantly higher than that in the control group (90.0% vs. 72.5%, χ2 = 4.02, P < 0.05). At 3, 6 and 12 months after treatment, recurrence rate in the study group was 2.5%, 15.0% and 30.0%, respectively, which were significantly lower than 17.5%, 35.0%, and 52.5% in the control group ( χ2 = 5.00, 4.26, 4.17, all P < 0.05). Conclusion:Five-element music therapy for the treatment of bipolar disorder can markedly alleviate depression and mania, restore patient's emotional state, improve clinical efficacy, and decrease recurrence rate.