Background: Long-term survival and high-quality life of patients with gliomas depends on the extent of resection (EOR) and the protection of functional white matter fibers. The navigation system provides precise positioning for surgery based on preoperative magnetic resonance imaging (MRI) but the precision decreases when intraoperative brain drift occurs. Ultrasound (US) can support real-time imaging and correct brain shift. The real-time US-MRI multimodal fusion virtual navigation system (UMNS) is a new technique for glioma surgery. In order to obtain a maximum EOR and functional protection, this study aimed to explore the feasibility, efficiency, and safety of real-time UMNS for glioma surgery, and to evaluate the benefit of the new application by UMNS presetting markers between the tumor and functional white matter fiber surgery. Methods: A retrospective analysis included 45 patients who underwent glioma surgery, 19 patients with only intraoperative US, and 26 patients with UMNS. A preoperative plan was made by 3D-slicer software based on preoperative MRI. This was combined with a reconstruction of diffusion tensor imaging (DTI) that designed the important locations as “warning points” between functional white matter fibers and tumor. Following patient registration, markers were injected into preset “warning points” under image-guided UMNS in order to give us a warning during surgery in case of postoperative function deficits. The operating time, volumetric assessment in glioma resection, and postoperative complications were evaluated and used to compared those surgeries using intraoperative US (iUS) with those surgeries using intraoperate MRI (iMRI) navigation. Results: A total of 45 patients underwent glioma surgery. Gross total removal (GTR) of iUS alone was achieved in 6 of 19 cases, while this was achieved in 22 of 26 cases with UMNS alone, demonstrating an improvement in rate of GTR from 31.58% to 84.62%, respectively. This may be attributable to the superior US image quality provided by UMNS. In 13 of 26 cases, there was improved image quality (from poor/moderate to moderate/good) with the aid of UMNS. In addition, the consistency of EOR of postoperative MRI evaluated by UMNS (92.31%) was higher than when using iUS alone (42.11%). The whole process of intraoperative scanning time and marker injection did not lead to a significant delay of the operating time compared to using iUS alone, and has been reported to be shorter than with iMRI as well. Furthermore, the percentage of postoperative morbidity in the UMNS group was lower than that in the iUS group (motor deficit: 11.54% vs. 42.11%; aphasia: P =3.85% vs. 31.58%, respectively). Conclusions: Real-time UMNS is an effective, timesaving technology that offers high quality intraoperative imaging. Injection markers between functional white matter fibers and tumor by UMNS can help to obtain a maximum EOR of glioma and functional protection postoperatively. The integration of iUS into the neuronavigation system offered quick and helpful intra-operative images.

Antitumor effect of a new nano-vector with miRNA-135a on malignant glioma Chaofeng Liang,1,* Weitong Sun,2,* Haiyong He,1,* Baoyu Zhang,1 Cong Ling,1 Bocheng Wang,1 Tengchao Huang,1 Bo Hou,1 Ying Guo1 1Department of Neurosurgery, 3rd Affiliated Hospital of Sun Yat-sen University, Sun Yat-sen University, Guangdong, China; 2The Pharmaceutical College of Jiamusi University, Jiamusi University, Jiamusi, China *These authors contributed equally to this work Introduction: MiR-135a is found to selectively induce apoptosis in glioma cell but not in normal neurons and glial cells. However, low transfection efficacy limits its application in vivo as other miRNAs. We prepared a new kind of nano-vector based on polyethylene glycol methyl ether (mPEG) and hyper-branched polyethylenimine (hy-PEI) in order to improve the miRNA delivery system into the glioma cells. Methods: The mPEG-g-PEI/miR-135a was constructed and detected by 1H NMR and FTIR analyses. Transmission electron microscope was utilized for its characteristics. Stability and release efficiency was assessed by electrophoresis. Biocompatibility was observed and analyzed through co-culture with astrocytes and malignant glioma cells (C6). Transfection rate was monitored by laser confocal microscopy and flow cytometry. The antitumor effect of mPEG-g-PEI/miR-135a to C6 was confirmed in vivo by MR scanning, pathology and survival curve. RT-PCR was used to assay transfection efficiency of mPEG-g-PEI/miR-135a in vitro and in vivo. And Western blotting was used to assess the expressions of the targeted proteins of miR-135a.Results: In this experiment, we found the optimal N/P ratio of mPEG-g-PEI/miR-135a was about 6 judged by Zeta potential, particle size and encapsulation ability. The stability of mPEG-g-PEI/miR-135a in serum and the release efficiency in acid(pH=5.0) of mPEG-g-PEI/miR-135a were simulated the environment in vivo and in tumor. The mPEG-g-PEI nano-vector was co-cultured with malignant glioma cell C6 and normal astrocytes in vitro and showed good biocompatibility evaluated by CCK8 assay. The cell experiments in vitro indicated that mPEG-g-PEI could significantly improve miR-135a transfection by enhancing uptake effect of both normal glial and glioma cells. Given the C6 implanted in situ model, we discovered that the mPEG-g-PEI/miR-135a could obviously increase the survival period and inhibit the growth of glioma confirmed by MRI and histochemistry. In addition, the transfection efficiency of mPEG-g-PEI was better than that of other transfection agents either in vitro or in vivo confirmed by RT-PCR. Moreover, the expressions of the targeted proteins of miR-135a were consistent with the in vitro results. Conclusion: These results suggest that mPEG-g-PEI is expected to provide a new effective intracellular miRNA delivery system with low toxicity for glioma therapy. Keywords: nano-vector, miRNA-135a, malignant glioma, gene delivery, Micro RNA

Background: Alzheimer’s disease (AD) is a progressive neurodegenerative disorder with no effective treatments available. There is growing evidence that cuproptosis contributes to the pathogenesis of this disease. This study developed a novel molecular clustering based on cuproptosis-related genes and constructed a signature for AD patients.Methods: The differentially expressed cuproptosis-related genes (DECRGs) were identified using the DESeq2 R package. The GSEA, PPI network, GO, KEGG, and correlation analysis were conducted to explore the biological functions of DECRGs. Molecular clusters were performed using unsupervised cluster analysis. Differences in biological processes between clusters were evaluated by GSVA and immune infiltration analysis. The optimal model was constructed by WGCNA and machine learning techniques. Decision curve analysis, calibration curves, receiver operating characteristic (ROC) curves, and two additional datasets were employed to confirm the prediction results. Finally, immunofluorescence (IF) staining in AD mice models was used to verify the expression levels of risk genes.Results: GSEA and CIBERSORT showed higher levels of resting NK cells, M2 macrophages, naïve CD4⁺ T cells, neutrophils, monocytes, and plasma cells in AD samples compared to controls. We classified 310 AD patients into two molecular clusters with distinct expression profiles and different immunological characteristics. The C1 subtype showed higher abundance of cuproptosis-related genes, with higher proportions of regulatory T cells, CD8⁺T cells, and resting dendritic cells. We subsequently constructed a diagnostic model which was confirmed by nomogram, calibration, and decision curve analysis. The values of area under the curves (AUC) were 0.738 and 0.931 for the external datasets, respectively. The expression levels of risk genes were further validated in mouse brain samples.Conclusion: Our study provided potential targets for AD treatment, developed a promising gene signature, and offered novel insights for exploring the pathogenesis of AD.

The discovery of intracranial lymphatic system provides a new theory about cerebrospinal fluid (CSF) circulation. In order to investigate the role of lymphatic drainage in hydrocephalus after intraventricular hemorrhage (IVH), three parts of experiments were performed. IVH model was produced by 200ul autologous blood injection into lateral ventricle of SD male rats. The ability of iron drainage through deep cervical lymph nodes(DCLNs) was confirmed by Perls prussian blue reactionin in partI. The partIIdemonstrated DCLNs-excised rats got a 19.6% higher incidence of hydrocephalus than DCLNs-preserved rats at day 28 after IVH. The partIII compared the ferritin level of area surrounding ventricles by Western blotting after IVH. Results indicated the DCLNs-excised group showed higher ferritin level than DCLNs-preserved group at day 3 after IVH. The whole experiments revealed the role of intracranial lymphatic system in discharging iron out of central nervous system (CNS) at acute phase of IVH, and proved the function in alleviating and avoiding hydrocephalus at chronic phase after IVH.

Deficits in astrocyte function contribute to major depressive disorder (MDD) and suicide, but the therapeutic effect of directly reactivating astrocytes for depression remains unclear. Here, specific gains and losses of astrocytic cell functions in the medial prefrontal cortex (mPFC) bidirectionally regulate depression-like symptoms. Remarkably, recombinant human Thrombospondin-1 (rhTSP1), an astrocyte-secreted protein, exerted rapidly antidepressant-like actions through tyrosine hydroxylase (Th)/dopamine (DA)/dopamine D2 receptors (D2Rs) pathways, but not dopamine D1 receptors (D1Rs), which was dependent on SH3 and multiple ankyrin repeat domains 3 (Shank3) in the mPFC. TSP1 in the mPFC might have potential as a target for treating clinical depression.