Zhongqiang Chen

Purpose To investigate the effectiveness of sagittal deformity angular distance ratio (S-DADR; sagittal Cobb angle divided by the distance, in centimeters, between the midpoint of the upper endplate of the upper end vertebrae and the midpoint of the lower endplate of the lower end vertebrae) in evaluating neurological risks in patients with post-tuberculous kyphosis (PTK). Methods A consecutive series of 102 patients with PTK who underwent corrective surgery between September 2010 and June 2024 were included. Measurements derived from preoperative radiographs were utilized to compute S-DADR. Potential risk factors of postoperative neurological deficits (PNDs) were assessed through univariate and multivariate analyses. Binary logistic regression was used to model the relationship between identified independent risk factors and PNDs, and the model was evaluated by the area under receiver operating characteristic curve (AUC). Results The mean age of the 102 patients was 49.5 ± 12.1 years, and the incidence of PNDs was 22.5%. Univariate logistic regression revealed a positive correlation between the incidence of PNDs and S-DADR (OR = 1.103, P = 0.003), preoperative sagittal Cobb angle (OR = 1.025, P = 0.007), number of vertebrae resected (OR = 1.661, P = 0.024), operative time (OR = 1.005, P = 0.048) and total instrumented vertebrae (OR = 1.391, P = 0.014). Multivariate logistic regression identified S-DADR (OR = 1.091, P = 0.025) as an independent factor of PNDs. Patients with S-DADR ≥ 21.4 exhibited a 30% incidence of PNDs, which increased to 50% with an S-DADR of 30.0. The overall predictive performance of S-DADR (AUC = 0.720) for PNDs was deemed satisfactory. Conclusions S-DADR can be used to quantify the severity of the deformity in patients with PTK, which is an independent risk factor for PNDs during corrective surgery. Patients with S-DADR ≥ 21.4 face a 30% incidence of developing PNDs after corrective surgery.

Oxeiptosis is a reactive oxygen species (ROS)-induced pathway of cell death. The involvement of circular RNAs (circRNAs) has been confirmed in the incidence and progression of intervertebral disc degeneration (IVDD). However, whether oxeiptosis occurs in IVDD and how circRNAs regulate oxeiptosis is still unclear. In this study, we discovered that oxeiptosis could be induced in nucleus pulposus cells (NPCs), and circFOXO3 was significantly upregulated after oxeiptosis induction. Transfection using circFOXO3 small interfering RNA (siRNA) significantly inhibited oxeiptosis in NPCs. Mechanistically, circFOXO3 upregulated acid-sensing ion channel subunit 1 (ASIC1) expression by functioning as a molecular sponge for miR-185-3p and miR-939-5p. Subsequent rescue experiments validated that circFOXO3 could regulate oxeiptosis in NPCs via the miR-185-3p/miR-939-5p-ASIC1 axis. Further research on ASIC1 functions indicated that this regulation was achieved by affecting the Calcium ion (Ca²⁺) influx mediated by ASIC1. A mouse IVDD model was established, and silencing circFOXO3 in vivo was found to inhibit IVDD development and the activation of the oxeiptosis-related pathway. Overall, circFOXO3 is one of the factors contributing to the progression of IVDD by mediating oxeiptosis.

Background Thoracic ossification of ligamentum flavum (TOLF) can be asymptomatic and progress insidiously. But, long-term follow-up results of clinical progression of TOLF are still unknown. Methods The clinical progression of 81 patients with TOLF at our center, followed for 10 to 11 (mean, 10.3) years from May 2010 to November 2021, were analyzed. Among them, 51 patients with thoracic myelopathy were caused by single- or multi-segment TOLF, and received partial TOLF resection (30 patients) or total TOLF resection (21 patients). The remaining 30 patients showed TOLF on imaging examinations, but TOLF was not the responsible compressing factor causing myelopathy and with no TOLF resection. The mJOA score (total 11 scores) and spinal operation were used to evaluate the clinical progression at follow-up. Results During the 10- to 11-year follow-up of 81 TOLF patients, 71 (87.7%) had no deterioration of neurological function, and 10 (12.3%) patients had deterioration of neurological function and had another spinal operation, including only 4 (4.9%) suffered thoracic myelopathy caused by the progression of TOLF; 6 (7.4%) for other spinal diseases: 2 (2.5%) had fall damage and acute spinal cord injury at the TOLF level; 2 (2.5%) had thoracic myelopathy caused by ossification of posterior longitudinal ligament (OPLL); 2 (2.5%) had cervical spondylosis and received cervical operation. Conclusions Most TOLF (87.7%) patients had no clinical progression and received no reoperations for TOLF in the ten-year dimension (mean, 10.3 years). Narrow spinal canal for TOLF increases the risk of traumatic paraplegia.

Background: Diffuse idiopathic skeletal hyperostosis (DISH) has been evaluated as a potential risk factor of poor surgical outcomes for lumbar spinal stenosis, whereas the influence of DISH on neuroimaging characteristics and postoperative prognosis of patients with thoracic myelopathy has not been established. Therefore, this study aimed to shed light on this issue. Methods: A monocentric study enrolled 167 eligible patients with thoracic ossification of ligamentum flavum (TOLF), who were followed up for at least 2 years. Clinico-radiological parameters and surgical outcomes were compared between the DISH+ and DISH− groups before and after propensity matching. Subgroup analysis was conducted to compare the functional outcomes between mild DISH (M-DISH) and moderately severe DISH (MS-DISH) groups. Results: Fifty-eight patients were diagnosed as DISH, and its prevalence was 34.7%. Patients with DISH presented with older age, more males, taller stature, heavier weight, more commonly diffuse-type TOLF (p < 0.05). The DISH group showed significantly worse recovery rate (RR) at the final follow-up before and after propensity matching (p < 0.01), and slightly lower preoperative VAS, higher postoperative VAS and lower VAS reduction, despite not reaching the significant differences. Subgroup analysis demonstrated that the M-DISH group was associated with the lower mJOA score (p = 0.01) and RR at the final follow-up (p = 0.001), and tended to present higher preoperative VAS than the MS-DISH group. Conclusions: DISH has a significant predisposition to the elderly males with diffuse-type TOLF. Although the presence of M-DISH might bring about a suboptimal surgical outcome, both DISH and non-DISH patients experienced good neurological function improvements and pain relief through thoracic posterior decompression.

Thoracic ossification of the ligamentum flavum (TOLF) is ectopic ossification of the spinal ligaments. Histologically, the development of TOLF can be described as the process of endochondral ossification. However, the underlying aetiology has not been completely clarified. In this investigation, the gene expression profile associated with leucine‐rich repeat‐containing G‐protein‐coupled receptors (LGR) and Wnt signalling pathway in the thoracic ligamentum flavum cells (TLFCs) of different ossification stages was analysed via RNA sequencing. We further confirmed the significant differences in the related gene expression profile by Gene Ontology (GO) enrichment analysis. LGR5 was first identified in primary human TLFCs during osteogenic differentiation. To evaluate the effect of LGR5 on osteogenic differentiation, LGR5 has been knocked down and overexpressed in human TLFCs. We observed that the knockdown of LGR5 inhibited the activity of Wnt signalling and attenuated the potential osteogenic differentiation of TLFCs, while overexpression of LGR5 activated the Wnt signalling pathway and increased osteogenic differentiation. Our results provide important evidence for the potent positive mediatory effects of LGR5 on osteogenesis by enhancing the Wnt signalling pathway in TOLF.

Background: Ossification of ligamentum flavum (OLF) is an insidious and debilitating heterotopic ossifying disease with etiological heterogeneity and undefined pathogenesis. Obese individuals predispose to OLF, whereas the underlying connections between obesity phenotype and OLF pathomechanism are not fully understood. Therefore, this study aims to explore distinct obesity-related genes and their functional signatures in OLF.Methods: The transcriptome sequencing data related to OLF were downloaded from the GSE106253 in the Gene Expression Omnibus (GEO) database. The obesity-related differentially expressed genes (ORDEGs) in OLF were screened, and functional and pathway enrichment analysis were applied for these genes. Furthermore, protein-protein interactions (PPI), module analysis, transcription factor enrichment analysis (TFEA), and experiment validation were used to identify hub ORDEGs. The immune infiltration landscape in OLF was depicted, and correlation analysis between core gene SOCS3 and OLF-related infiltrating immune cells (OIICs) as well as 5mC/m6A modifiers in OLF was constructed.Results: Ninety-nine ORDEGs were preliminarily identified, and functional annotations showed these genes were mainly involved in metabolism, inflammation, and immune-related biological functions and pathways. Integrative bioinformatic algorithms determined a crucial gene cluster associated with inflammatory/immune responses, such as TNF signaling pathway, JAK-STAT signaling pathway, and regulation of interferon-gamma-mediated signaling. Eight hub ORDEGs were validated, including 6 down-regulated genes (SOCS3, PPARG, ICAM-1, CCL2, MYC, and NT5E) and 2 up-regulated genes (PTGS2 and VEGFA). Furthermore, 14 differential OIICs were identified by ssGSEA and xCell, and SOCS3 was overlapped to be the core gene, which was associated with multiple immune infiltrates (dendritic cells, macrophage, and T cells) and six m6A modifiers as well as four 5mC regulators in OLF. Reduced SOCS3 and FTO expression and up-regulated DNMT1 level in OLF were validated by Western blotting.Conclusion: This study deciphered immune/inflammatory signatures of obesity-related gene clusters for the first time, and defined SOCS3 as one core gene. The crosstalk between 5mC/m6A methylation may be a key mediator of SOCS3 expression and immune infiltration. These findings will provide more insights into molecular mechanisms and therapeutic targets of obesity-related OLF.

Background: Obese individuals predispose to ossification of ligamentum flavum (OLF), whereas the underlying connections between obesity phenotype and OLF pathomechanism are not fully understood, especially during early life. This study aimed to explore obesity-associated genes and their functional signatures in OLF. Methods: Gene microarray expression data related to OLF were downloaded from the GSE106253 dataset in the Gene Expression Omnibus (GEO) database. The potential obesity-related differentially expressed genes (ORDEGs) in OLF were screened. Then, gene-ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were applied for these genes. Furthermore, protein-protein interactions (PPI) were used to identify hub ORDEGs, and Metascape was used to further verify the key signaling pathways and immune-related function signatures of hub ORDEGs. Finally, correlation analysis of hub ORDEGs and identified OLF-related infiltrating immune cells (OIICs) was constructed to understand the possible mechanical link among obesity, immune response and OLF. Results: OLF-related differentially expressed genes and 2051 obesity-related genes from four databases were intersected to obtain 99 ORDEGs, including 54 upregulated and 55 downregulated genes. GO and KEGG analysis revealed that these genes were mainly involved in metabolism, inflammation and immune-related biological functions and pathways. A PPI network was established to determine 14 hub genes (AKT1, CCL2, CCL5, CXCL2, ICAM1, IL10, MYC, PTGS2, SAA1, SOCS1, SOCS3, STAT3, TNFRSF1B and VEGFA). The co-expression network demonstrated that this module was associated with cellular response to biotic stimulus, regulation of inflammatory response, regulation of tyrosine phosphorylation of STAT protein. Furthermore, Metascape functional annotations showed that hub genes were mainly involved in receptor signaling pathway via JAK-STAT, response to TNF and regulation of defense response, and their representative enriched pathways were TNF, adipocytokine and JAK-STAT signaling pathways. Subgroup analysis indicated that T cell activation might be potential immune function processes involved, and correlation analysis revealed that cDCs, memory B-cells and preadipocytes were highly correlated infiltrating immune cells. Conclusions: Our study deciphered individualized obesity-associated gene signature for the first time, which may facilitate exploring the underlying cellular and molecular pathogenesis and novel therapeutic targets of obesity-related early-onset OLF.

Purpose Genetic factors play a crucial role in thoracic ossification of the ligamentum flavum (TOLF). This study aimed to better understand the association between single nucleotide polymorphisms (SNP) in functional regions of the collagen VI, alpha 1 gene (COL6A1) and TOLF, and to confirm COL6A1 as a TOLF susceptibility gene. Methods Ten tag SNPs in COL6A1 were genotyped using the SNaPshot assay, and allele and genotype frequencies were compared between TOLF patients and control individuals. The function of SNPs associated with disease was studied. For COL6A1 promoter SNPs, the transcriptional activity of each haplotype was determined by luciferase reporter assays. For COL6A1 exonic SNPs, the effect of nucleotide substitutions on COL6A1 expression was determined by western blotting. COL6A1 mRNA expression in ligamentum flavum tissues from TOLF patients with different genotypes was examined using reverse transcription real-time PCR. Results Four SNPs were associated or possibly associated with TOLF, with higher pathogenic allele and genotype frequencies seen in TOLF patients compared with controls. The rs17551710/rs7671-GG/GG genotype appeared to be related to disease severity. Nucleotide substitutions at rs17551710 and rs7671 increased COL6A1 transcriptional activity and nucleotide substitutions at rs1053312 and rs13051496 increased COL6A1 protein expression. COL6A1 mRNA expression was significantly up-regulated in individuals with rs17551710/rs7671-GG/GG and rs1053312/rs13051496-AA+AG/CC genotypes compared with other genotypes. Conclusion SNPs in the COL6A1 promoter and exonic regions are associated with TOLF in the Chinese Han population, and lead to up-regulated COL6A1 expression. We confirmed COL6A1 as a TOLF susceptibility gene that may be involved in TOLF pathology.

Purpose: Ossification of the ligamentum flavum (OLF) is a multifactorial disease characterized by an insidious and debilitating process of abnormal bone formation in ligamentum tissues. However, its definite pathogenesis has not been fully elucidated. Potential links between the immune system and various forms of heterotopic ossification have been discussed for many years, whereas no research investigated the immune effects on the initiation and development of OLF. Therefore, we attempt to shed light on this issue. Methods: A series of bioinformatic algorithms were integrated to evaluate the immune score and the immunocyte infiltration patterns between OLF and normal samples, screen OLF-related and immune-related differentially expressed genes (OIDEGs), and analyze their biological functions. Correlation analysis inferred OIDEGs-related differentially expressed lncRNAs (OIDELs) and infiltrating immune cells (OIICs) to construct an immunoregulatory network. Results: Differential immune score and immune cell infiltration were determined between two groups, and 10 OIDEGs with diverse biological function annotations were identified and verified. A lncRNA-gene-immunocyte regulatory network further revealed 10 OIDEGs, 41 OIDELs and 7 OIICs that were highly correlated. Among them, CD1E and STAT3 were predicted as hub genes whether at the expression level or interaction level. cDCs emerged as having the most prominent differences and the highest degree of connectivity. FO393414.3, AC096734.1, LINC01137 and DLX6-AS1 with the greatest number of OIDEGs were thought to be more likely to participate in immunoregulation of OLF. Conclusion: This is the first research to preliminarily elucidate OLF-related immunocyte infiltration landscape and immune-associated transcriptome signatures based on bioinformatic strategies and real-world data, which may provide compelling insights into the pathogenesis and therapeutic targets of OLF.