Combination of machine learning-based bulk and single-cell genomics reveals necroptosis-related molecular subtypes and immunological features in autism spectrum disorder.

Background: Necroptosis is a novel form of controlled cell death that contributes to the progression of various illnesses. Nonetheless, the function and significance of necroptosis in autism spectrum disorders (ASD) remain unknown and require further investigation. Methods: We utilized single-nucleus RNA sequencing (snRNA-seq) data to assess the expression patterns of necroptosis in children with autism spectrum disorder (ASD) based on 159 necroptosis-related genes. We identified differentially expressed NRGs and used an unsupervised clustering approach to divide ASD children into distinct molecular subgroups. We also evaluated immunological infiltrations and immune checkpoints using the CIBERSORT algorithm. Characteristic NRGs, identified by the LASSO, RF, and SVM-RFE algorithms, were utilized to construct a risk model. Moreover, functional enrichment, immune infiltration, and CMap analysis were further explored. Additionally, external validation was performed using RT-PCR analysis. Results: Both snRNA-seq and bulk transcriptome data demonstrated a greater necroptosis score in ASD children. Among these cell subtypes, excitatory neurons, inhibitory neurons, and endothelials displayed the highest activity of necroptosis. Children with ASD were categorized into two subtypes of necroptosis, and subtype2 exhibited higher immune activity. Four characteristic NRGs (TICAM1, CASP1, CAPN1, and CHMP4A) identified using three machine learning algorithms could predict the onset of ASD. Nomograms, calibration curves, and decision curve analysis (DCA) based on 3-NRG have been shown to have clinical benefit in children with ASD. Furthermore, necroptosis-based riskScore was found to be positively associated with immune activation. Finally, RT-PCR demonstrated differentially expressed of these four NRGs in human peripheral blood samples. Conclusion: A comprehensive identification of necroptosis may shed light on the underlying pathogenic process driving ASD onset. The classification of necroptosis subtypes and construction of a necroptosis-related risk model may yield significant insights for the individualized treatment of children with ASD.

Three-Dimensional Finite Element Analysis of L4-5 Degenerative Lumbar Disc Traction under Different Pushing Heights.

Objective: To study and analyze the changes of intervertebral foramen height and area of the degenerative L4-5 intervertebral disc under different pushing heights by the finite element method. Methods: CT and MRI images of T12-S1 segments were obtained from a healthy volunteer who met the inclusion criteria. A DR machine was used to capture images of the lumbar lateral section before and after simultaneous pushing of the L4 and L5 spinous processes by manipulation called Daogaijinbei, and the measurement showed that the displacement changes of L4 and L5 were both approximately 10 cm, so the pushing height was set at 0-10 cm. A three-dimensional finite element model of the entire normal lumbar spine was established using Mimics 16.0, Geomagic Studio 2014, Hypermesh 13.0, MSC.Patran 2012, and so on. The disc height and nucleus area of the lumbar disc of the normal entire lumbar disc model were adjusted to establish models of the L4-5 disc with mild, moderate, and severe degeneration. Changes of disc height and area of the L4-5 degenerative intervertebral disc under different pushing heights were calculated. Results: The size of the L4-5 intervertebral foramen was analyzed from the height and area of the intervertebral foramen, and the results showed the following: (1) as for the normal lumbar disc and a lumbar of the L4-5 disc with mild and moderate degeneration, the height of the L4-5 intervertebral foramen and its area both increased during pushing between 0 and 8 cm. After the pushing height reached 8 cm, the height and area of the L4-5 intervertebral foramen gradually became stable; (2) as for the L4-5 disc with severe degeneration, during the process of pushing, the height and area of the L4-5 intervertebral foramen increased slightly, but this change was not obvious. Conclusions: After the spinal manipulation, the sizes of the L4-5 intervertebral foramen of the L4-5 disc with mild and moderate degeneration were significantly larger than those before pushing; in contrast, the size of L4-5 intervertebral foramen of the L4-5 disc with severe lumbar degeneration was not significantly changed.