Objective:To evaluate the topological alterations of resting-state brain networks in patients with idiopathic generalized epilepsy with generalized tonic-clonic seizure (IGE-GTCS) using minimum spanning tree (MST) based on graph theoretic analysis, and to further analyze the relationships between topological features, duration, and antiepileptic drug response.Methods:This study was a cross-sectional study. Retrospectively, 75 IGE-GTCS patients and 37 healthy controls (HC) who underwent brain MR imaging at the Affiliated of Nanjing University Medical School Drum Tower Hospital from January 2013 to December 2020 were enrolled. IGE-GTCS patients were grouped into well-controlled subgroup (WC; n=55) and drug-resistant subgroup (DR; n=20) according to their response to antiepileptic drugs. Firstly, the time series correlations between 116 regions of the whole brain of each subject were calculated to construct functional connectivity matrices. For each functional connectivity matrix, the Kruskal algorithm was used to MST, and the topological metrics of each MST were calculated, including leaf fraction, tree hierarchy, and diameter. The comparison of MST topological metrics between the two groups was performed using two-sample t-test. Pearson correlation analysis was used to calculate the correlation between disease duration and MST metrics in the WC subgroup and the DR subgroup. Results:Compared with the HC group, the MST leaf fraction ( t=2.27, P=0.025) increased in the IGE-GTCS patient group, and the diameter decreased ( t=-2.24, P=0.027), there was no statistically significant difference in tree hierarchy between IGE-GTCS patient group and HC group ( t=0.98, P=0.328). The MST leaf fraction ( t=-2.39, P=0.019) and tree hierarchy ( t=-2.24, P=0.027) in the WC subgroup was decreased compared with the DR subgroup, while there was no statistically significant difference in diameter between WC subgroup and DR subgroup ( P=0.093). The correlation analysis showed the MST diameter in WC subgroup was significantly correlated with disease duration ( r=0.452, P<0.001), while the MST diameter in DR subgroup was not significantly correlated with disease duration ( r=-0.062, P=0.847). Conclusions:Patients with IGE-GTCS exhibit specific alterations in the global topology of brain network, characterized by increased centralization and efficiency. The effective antiepileptic drug treatment is associated with a recovery of brain network abnormalities.

Mental state is an important part of the normal life activities of the human body, and it is also the most external expression and the most easily obtained information of the physical condition. The normal activities of the mind depend on the normal operation of the viscera, qi, and blood, and are a unified whole that prospers together and suffers together. The theory of the five-spirit-viscera in the Yellow Emperor’s Inner Classic revealed that the normal mental activities of the human body were dominated by the five internal organs, that is, the five internal organs were the body and the five spirits were the function. And it highlighted the viewpoint that the five internal organs store the spirits and are actually one. The heart governs the spirit and belongs to the four internal organs. On this basis, Synopsis of Golden Chamber used the internal organs to diagnose and treat mental diseases, integrating the theory of the five spirits into it, forming a unique method of diagnosis and treatment with the heart as the leading factor and regulating the qi and blood of the four internal organs. It identified the pathogenesis of diseases such as pathogenic crying, lily disease, and hysteria from five levels: heart deficiency and weak qi, heart-lung disharmony, heart-liver disharmony, the heart of the loss of the spleen nourishment, and disharmony between heart and kidney. The treatment was mainly to replenish the deficiency of the viscera and eliminate the pathogens, reflecting the characteristics of regulating the mind and calming the four internal organs. This unique view on diagnosis and treatment has profoundly influenced the diagnosis and treatment theories of mental illnesses by later doctors, and is of great significance to the current clinical treatment of such illnesses.

Objective:To evaluate the topological alterations of resting-state brain networks in patients with idiopathic generalized epilepsy with generalized tonic-clonic seizure (IGE-GTCS) using minimum spanning tree (MST) based on graph theoretic analysis, and to further analyze the relationships between topological features, duration, and antiepileptic drug response.Methods:This study was a cross-sectional study. Retrospectively, 75 IGE-GTCS patients and 37 healthy controls (HC) who underwent brain MR imaging at the Affiliated of Nanjing University Medical School Drum Tower Hospital from January 2013 to December 2020 were enrolled. IGE-GTCS patients were grouped into well-controlled subgroup (WC; n=55) and drug-resistant subgroup (DR; n=20) according to their response to antiepileptic drugs. Firstly, the time series correlations between 116 regions of the whole brain of each subject were calculated to construct functional connectivity matrices. For each functional connectivity matrix, the Kruskal algorithm was used to MST, and the topological metrics of each MST were calculated, including leaf fraction, tree hierarchy, and diameter. The comparison of MST topological metrics between the two groups was performed using two-sample t-test. Pearson correlation analysis was used to calculate the correlation between disease duration and MST metrics in the WC subgroup and the DR subgroup. Results:Compared with the HC group, the MST leaf fraction ( t=2.27, P=0.025) increased in the IGE-GTCS patient group, and the diameter decreased ( t=-2.24, P=0.027), there was no statistically significant difference in tree hierarchy between IGE-GTCS patient group and HC group ( t=0.98, P=0.328). The MST leaf fraction ( t=-2.39, P=0.019) and tree hierarchy ( t=-2.24, P=0.027) in the WC subgroup was decreased compared with the DR subgroup, while there was no statistically significant difference in diameter between WC subgroup and DR subgroup ( P=0.093). The correlation analysis showed the MST diameter in WC subgroup was significantly correlated with disease duration ( r=0.452, P<0.001), while the MST diameter in DR subgroup was not significantly correlated with disease duration ( r=-0.062, P=0.847). Conclusions:Patients with IGE-GTCS exhibit specific alterations in the global topology of brain network, characterized by increased centralization and efficiency. The effective antiepileptic drug treatment is associated with a recovery of brain network abnormalities.

Developing new therapeutic agents for cancer immunotherapy is highly demanding due to the low response ratio of PD-1/PD-L1 blockade in cancer patients. Here, we discovered that the novel immune checkpoint VISTA is highly expressed on a variety of tumor-infiltrating immune cells, especially myeloid derived suppressor cells (MDSCs) and CD8⁺ T cells. Then, peptide C1 with binding affinity to VISTA was developed by phage displayed bio-panning technique, and its mutant peptide VS3 was obtained by molecular docking based mutation. Peptide VS3 could bind VISTA with high affinity and block its interaction with ligand PSGL-1 under acidic condition, and elicit anti-tumor activity in vivo. The peptide DVS3-Pal was further designed by d-amino acid substitution and fatty acid modification, which exhibited strong proteolytic stability and significant anti-tumor activity through enhancing CD8⁺ T cell function and decreasing MDSCs infiltration. This is the first study to develop peptides to block VISTA/PSGL-1 interaction, which could act as promising candidates for cancer immunotherapy.

MicroRNAs(miRNAs)are important regulators of gene expression.The large-scale detection and profiling of miRNAs have been accelerated with the development of high-throughput small RNA sequencing(sRNA-Seq)techniques and bioinformatics tools.However,generating high-quality comprehensive miRNA annotations remains challenging due to the intrinsic complexity of sRNA-Seq data and inherent limitations of existing miRNA prediction tools.Here,we present iwa-miRNA,a Galaxy-based framework that can facilitate miRNA annotation in plant species by combining computational analysis and manual curation.iwa-miRNA is specifically designed to generate a comprehensive list of miRNA candidates,bridging the gap between already annotated miRNAs provided by public miRNA databases and new predictions from sRNA-Seq datasets.It can also assist users in selecting promising miRNA candidates in an interactive mode,contributing to the accessibility and reproducibility of genome-wide miRNA annotation.iwa-miRNA is user-friendly and can be easily deployed as a web application for researchers without programming experience.With flexible,interactive,and easy-to-use features,iwa-miRNA is a valu-able tool for the annotation of miRNAs in plant species with reference genomes.We also illustrate the application of iwa-miRNA for miRNA annotation using data from plant species with varying genomic complexity.

Objective:To compare the effectiveness between three methods for purifying the immunoglobulin of egg yolk(IgY) which are polyethylene glycol (PEG) method, chloroform extraction method and chloroform / PEG method, and to provide basis for obtaining the batch of IgY.Methods:The inactivated vaccine of Vibrio parahemolyticus (V. parahemolyticus) was prepared and the hens were immunized by multi-point intramuscular injection.The eggs were collected and the IgY was purified by PEG method, chloroform extraction method and chloroform/PEG method.The protein extraction rate, the IgY titer and the purity of the antibody which purified by different methods were detected.Furthermore, the operation process, cost and safety of the three methods were analyzed.Results:The protein contents of the extraction belonging three methods from high to low in turn were chloroform extraction method, chloroform/PEG method, and PEG method.There was no significant difference in the antibody titer between three methods, and the tiler of chloroform extraction method was slightly high.The purities of purified antibody from high to low in turn were PEG method, chloroform/PEG method and chloroform method.The PEG method had better security but relatively lower extraction efficiency and higher cost.The chloroform/PEG method had high extraction efficiency and good antibody purity.Conclusion:The PEG method is suitable for a small amount of extraction in the laboratory.The chloroform/PEG method is appropriate for extracting the high quality IgY in a batch as it has high extraction efficiency and good antibody purity.