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 detect the changes of autophagic flux at different stages after oxygen⁃glucose deprivationreoxygenation (OGD/R) with several highly sensitive methods. Methods : Primary cortical neurons after oxygen deprivation of sugar after reoxygenation (OGD/R) were divided into the experimental OGD/R group and OGD/R + bafilomycinA1 (BafA1) group , using an RFP⁃GFP series fluorescent tags LC3 gene transfection detection cytolysosome and fusion of lysosomes , transmission electron microscope (TEM) observation the ultrastructure of autophagy , p62/SQSTM1 combining LC3 protein to flip the experimental testing p62 and LC3 protein quantitative , p62 immune staining observing the distribution and content. Results : Under fluorescence microscope , the ratio of autophagy lysosome to autophagosome increased significantly in OGD/R group , and the changes of autophagy structure in different stages could be observed in TEM. The ratio of soluble p62 and LC3 Ⅱ/ Ⅰ reflected the activation of autophagic flux , and p62 was mainly distributed in BafA1 group after fluorescence staining. Conclusion Each method has its own advantages , and different methods and indicators can accurately reflect the specific changes of autophagic flux in different stages after neuronal OGD/R. Mastering and applying these methods can effectively explore central nervous system diseases from the perspective of autophagy.

Activation of the heart normally begins in the sinoatrial node (SAN). Electrical impulses spontaneously released by SAN pacemaker cells (SANPCs) trigger the contraction of the heart. However, the cellular nature of SANPCs remains controversial. Here, we report that SANPCs exhibit glutamatergic neuron-like properties. By comparing the single-cell transcriptome of SANPCs with that of cells from primary visual cortex in mouse, we found that SANPCs co-clustered with cortical neurons. Tissue and cellular imaging confirmed that SANPCs contained key elements of glutamatergic neurotransmitter system, expressing genes encoding glutamate synthesis pathway (Gls), ionotropic and metabotropic glutamate receptors (Grina, Gria3, Grm1 and Grm5), and glutamate transporters (Slc17a7). SANPCs highly expressed cell markers of glutamatergic neurons (Snap25 and Slc17a7), whereas Gad1, a marker of GABAergic neurons, was negative. Functional studies revealed that inhibition of glutamate receptors or transporters reduced spontaneous pacing frequency of isolated SAN tissues and spontaneous Ca

Objective In this study,the clinical data of single-center acute posterior circulation stroke patients were analyzed to compare the efficacy of TOAST classification for large artery atherosclerosis (LAA) and cardioembolism (CE) stroke patients after intravascular treatment difference.Methods Patient data comes from the advanced stroke center of the First Affiliated Hospital of Shihezi University School of Medicine,Xinjiang,China.According to the TOAST classification,the 98 people included were divided into LAA group and CE group.The mTICI score was used to evaluate vascular recanalization,the 90 day mRS score was used to evaluate the prognosis,the incidence of intracranial hemorrhage conversion within 72 hours and 90 day mortality Assess the safety of intravascular treatment.Results Among the 98 patients included in the study,79 were LAA stroke patients and 19 were CE patients.Compared with CE patients,LAA stroke patients are younger (60.08±12.64 vs 71.05±10.42,P=0.001),have a higher prevalence of hyperlipidemia (55.7% vs 22.1%,P=0.014),and preoperative systolic blood pressure higher (149.25±21.24 vs 134.79±22.83,P=0.010),lower NIHSS score (17.3 vs 22.2,P=0.025),better collateral circulation (ASITN/SIR score of 2~4 ratio,63.3% vs 21.1 %,P=0.002),the angioplasty rate is higher (31.6% vs 5.3%,P=0.040).However,the EVT recanalization rate (87.5% vs 84.2%,P=1.000) of the two groups of patients,the 90 day neurological function recovery rate (mRS score 0~2) (58.2% vs 36.8%,P=0.093),The incidence of postoperative intracranial hemorrhage transformation (1.3% vs 0%,P=1.000) and the 90 day stroke-related mortality (20.3% vs 42.1%,P=0.091) were not statistically significant.Conclusion s Although there were differences in baseline clinical data and clinical characteristics of EVT treatment between LAA and CE in posterior circulation,there was no significant difference in prognosis.