Jiandang Shi

Objective: To characterize the clinical, radiological, and microbiological features of pyogenic spondylitis (PS) and to describe outcomes observed within a severity-stratified real-world management pathway integrating antimicrobial therapy, stabilization, debridement, and fusion according to structural instability and neurological risk. Methods: We conducted a retrospective observational cohort study of 149 patients with PS treated at Ningxia Medical University General Hospital between January 2015 and December 2023. Patients were retrospectively grouped according to baseline structural severity, neurological status, and the management strategy actually used in routine care: Group A, antimicrobial therapy plus external immobilization (n = 48); Group B, posterior fixation and fusion plus antimicrobial therapy (n = 57); and Group C, posterior fixation with debridement and fusion via posterior and/or anterior approaches plus antimicrobial therapy (n = 44). Clinical characteristics, microbiological findings, inflammatory markers, VAS, ODI, Cobb angle, perioperative variables, complications, and endpoint-defined recovery were analyzed. Principal-component analysis and adjusted response models were prespecified as exploratory analyses. Results: The three groups represented distinct severity strata, ranging from structurally stable disease to destructive disease with neurological compromise. Final diagnostic adjudication identified 73 microbiologically confirmed cases, whereas the prespecified patient-level yield analysis was limited to a 66-case interpretable pretreatment subset, within which positivity was lower in Group C than in Groups A and B (40.0% vs 82.4% and 84.2%, respectively; P = 0.001). At the specimen level, 95 bacterial isolate records were summarized, and S. aureus was the predominant isolate (48/95). All groups showed improvement in inflammatory, pain, functional, and radiological measures over follow-up, although the tempo of recovery differed across severity-matched management strata. Exploratory adjusted analyses were directionally consistent with these descriptive patterns but should be interpreted cautiously. Conclusions: In this retrospective real-world cohort, a severity-stratified management pathway for PS was feasible and showed coherent microbiological, inflammatory, functional, and radiological recovery patterns within each clinical stratum. These findings support pragmatic escalation according to structural instability and neurological risk, but they should not be interpreted as comparative evidence of treatment superiority.

Background: Spinal tuberculosis (STB) and brucellar spondylitis (BS) show substantial overlap in clinical and imaging manifestations, but their local immunopathological programs and corresponding cellular states have not been systematically compared. This study integrated histopathology and single-cell transcriptomics to compare lesion-level differences between STB and BS and to characterize macrophage states associated with the foamy cell reaction. Methods: This retrospective case-control study included 80 STB cases and 80 BS cases treated between January 2020 and December 2024. H&E-stained lesion tissues from the full surgical cohort were reviewed for comparative histopathology. Fresh lesion nucleus pulposus tissue from six representative cases (STB = 3, BS = 3) underwent scRNA-seq re-analysis with cell-type annotation, myeloid subclustering, functional module scoring, GSVA, and CellChat analysis. Dual immunofluorescence for CD68 paired with CYP27A1, PLIN2, or PPARγ was used as tissue-level validation. The scRNA-seq workflow focused on the monocyte-macrophage lineage to evaluate lipid-remodelled states associated with the foamy cell reaction. Tissue-level immunofluorescence and supplementary statistical summaries were interpreted as orthogonal support for the transcriptomic findings rather than as stand-alone diagnostic criteria. Results: Histopathological comparison showed that STB and BS were both dominated by chronic inflammation but exhibited distinct lesion patterns. STB was characterized mainly by tuberculous nodules, caseous necrosis, Langhans giant cells, and granulomatous inflammation, whereas BS more often showed plasma cell infiltration, foamy cell reaction, eosinophil infiltration, and fibrotic repair. These between-group differences were statistically significant ( P < 0.05 or P < 0.01, as appropriate). scRNA-seq yielded 67,274 high-quality cells and identified nine major cell types. Re-clustering of the Mono_Macrophage_DC lineage identified 15 subclusters, among which Macro4 was relatively enriched in BS. Rather than showing the highest value for every broad foam-cell-related metric, Macro4 was supported by convergent evidence including relative enrichment in BS, coordinated expression of PPARG, APOE, APOC1, PLIN2, and CYP27A1, and higher lipid-droplet-formation and cholesterol-efflux-related programs in BS_Macro4. Dual immunofluorescence showed higher CD68-normalized overlap fractions for CYP27A1, PLIN2, and PPARγ in BS lesion tissues than in STB lesion tissues ( P < 0.05). CellChat analysis further suggested that Macro4 contributed relatively more to the HGF-MET signalling context in BS, although these ligand-receptor relationships should be interpreted as transcriptome-inferred candidate interactions. Conclusions: STB and BS exhibit distinct local immunopathological programs. STB is characterized by a granulomatous-necrotizing inflammatory pattern, whereas BS more often shows a chronic inflammatory and tissue-repair-associated lesion background with a more prominent foamy cell reaction. Macro4 is best interpreted as a BS-associated lipid-remodelled macrophage state supported by convergent histological and transcriptomic evidence. These findings provide a lesion-level cellular framework for further pathological and mechanistic study rather than a validated stand-alone diagnostic cutoff.

Background: Spinal tuberculosis (STB) and Brucella spondylitis (BS) are the two most common infectious diseases of the spine. Despite overlapping clinical features, their immunopathological mechanisms remain poorly defined. Methods: We performed integrated transcriptomic profiling of infected nucleus pulposus tissues from STB and BS patients, combining single-cell RNA sequencing (STB n=3; BS n=3) with bulk RNA sequencing (STB n=7; BS n=5). Single-cell analysis resolved immune heterogeneity and T-cell subset architecture, whereas bulk RNA-seq defined global transcriptional signatures. Cross-platform integration identified concordant differentially expressed genes and shared pathways. AUCell was applied to quantify T-cell functional programmes. Pseudotime trajectory reconstruction, regulon inference, and cell-cell communication analyses were performed to delineate immune circuit differences. Results: A total of 67,274 high-quality cells were analysed, identifying nine major cell populations, with T cells as a dominant immune component. STB tissues were enriched in inflammatory cells, including monocytes/macrophages, neutrophils, and osteoclasts, whereas BS showed higher proportions of B and T cells. In STB, CD4 T cells were skewed toward IL26-CD4 T, Cytotoxic CD4 T, Proliferating CD4 T, and Treg subsets, while CD8 T cells were enriched in IFNG-AS1-CD8 T and exhaustion-like populations, reflecting sustained IFN-driven activation with proliferative and apoptotic stress. In contrast, BS exhibited expansion of GZMB-CTLs and IFNG-CTLs subsets, consistent with a focused cytotoxic programme with preserved memory features. Integrated single-cell and bulk analyses demonstrated convergence of overlapping genes on TCR, NF-κB, cytokine signalling, and osteoclast differentiation pathways. Functional scoring and pseudotime analysis indicated broader Th1/Th17 activation and progression toward terminal exhaustion in STB, whereas BS T cells maintained intermediate effector–memory states. Regulatory network analysis revealed a STAT1/IRF-dominated interferon axis in STB and an IRF4/EOMES/RFX3-centred effector–memory programme in BS. Cell-cell communication mapping showed an MHC-II-CD4 axis predominating in STB and an MHC-I-CD8 axis in BS. Despite divergent immune circuits, both conditions converged on osteoclast differentiation pathways, suggesting immune–bone metabolic coupling as a shared mechanism of vertebral destruction. Conclusions: STB and BS exhibit pathogen-specific T-cell immune circuits that converge on a common osteo-immune pathway, providing mechanistic insight and potential targets for host-directed therapy.

Many studies have been published on the characteristics of spinal tuberculosis, but not yet on the risk factors for complications after complete debridement. This study aimed to investigate the risk factors of postoperative complications in patients with thoracolumbar spinal tuberculosis (TB) after complete debridement. The clinical data of patients with thoracolumbar spinal TB after complete debridement admitted to General Hospital of Ningxia Medical University from January 2013 to December 2021 were included in this retrospective study. Patients were included if they had complete clinical data and a minimum follow-up duration of 1 year to ensure adequate assessment of postoperative complications. The study cohort was divided into 2 groups, including complication group and non-complication group according to the presence or absence of postoperative complications. The clinical characteristics of thoracolumbar spinal TB patients who developed postoperative complications were evaluated, and risk factors were analyzed by using univariate and binary multivariate logistic regression analysis. A total of 571 patients were included in this study: 92 patients with complications and 479 patients without complications. The results of the univariate analysis and multivariate binary logistic regression analysis showed that preoperative albumin P = .019), the number of diseased segments ≥3 (OR = 2.072; CI, 1.183–3.629, P = .011), anemia (OR = 1.691; CI, 1.047–2.731, P = .032) and drug resistance (OR = 1.768; CI, 1.011–3.091, P = .046) were independent risk factors for postoperative complications in patients with thoracolumbar spinal TB after complete debridement. Our findings support that the level of preoperative serum albumin, number of diseased segments, anemia, and drug-resistant TB are independent risk factors for postoperative complications in patients with thoracolumbar spinal TB after complete debridement. Therefore, improving preoperative serum albumin level, correcting anemia, actively preventing and monitoring drug-resistance may effectively reduce the risk of postoperative complications in patients with thoracolumbar spinal TB after complete debridement.

Epidemiological and clinical analyses of Brucellar spondylitis (BS) are crucial for public health authorities to enhance disease surveillance and optimize case management strategies. However, clinical epidemiological research on BS remains limited. This study analyzed the clinical and epidemiological characteristics of hospitalized BS patients at a hospital in Ningxia, aiming to provide evidence to support region-specific strategies for BS prevention and control. We retrospectively collected demographic profiles, epidemiological characteristics, clinical presentations, laboratory/imaging findings, therapeutic regimens, and prognostic outcomes of BS inpatients admitted to the orthopedic department of a hospital in Ningxia from January 2012 to December 2023. A total of 821 patients with BS were included, consisting of 564 males (68.7%; 564/821) and 257 females (31.3%; 257/821), with a median age of 55 years (range: 18–88 years). Among all patients (n = 821), 466 (56.76%) were agricultural or pastoral workers, and 578 (70.4%) had a documented history of contact with cattle or sheep. Low back pain was the most common presenting symptom, reported in 779 cases (94.88%; 779/821). The combined positive rate of the Rose Bengal Plate Test (RBPT) and the Standard Agglutination Test (SAT) in serological testing was 90.99% (747/821). Blood cultures were performed in 618 patients, of whom 217 (35.11%; 217/618) yielded positive results. Imaging examinations revealed that the lumbar spine was the most frequently involved region (59.81%; 491/821), with the L4–L5 segment being the most commonly affected (20.83%; 171/821). Single-segment involvement was observed in 668 patients (81.37%; 668/821), single-vertebra involvement in 44 patients (5.36%; 44/821), and multi-segment or trans-segment involvement in 109 patients (13.28%; 109/821). Of the 465 patients who underwent surgical treatment, the most common procedure was posterior fixation combined with lesion debridement, performed in 215 cases (46.24%; 215/465). Among the 754 patients who completed follow-up, the most frequently used antibiotic regimen was doxycycline plus rifampicin (60.05%; 493/754). Relapse occurred in 26 patients during follow-up (3.45%; 26/754). Brucellar spondylitis in Ningxia exhibits a temporal pattern aligned with regional brucellosis trends. Strengthening imaging-based diagnosis, clinical evaluation, and targeted follow-up of high-risk populations may help reduce recurrence and improve disease control. Not applicable.

Osteosarcoma, a primary malignant bone tumor predominantly affecting children and adolescents, exhibits a dismal prognosis for patients with metastatic or recurrent disease, characterized by low five-year survival rates. This poor outcome is largely attributed to the incomplete understanding of the molecular mechanisms governing osteosarcoma aggressiveness and metastasis, coupled with a paucity of effective early diagnostic biomarkers and targeted therapeutic strategies. Consequently, elucidating the critical molecular events driving osteosarcoma pathogenesis and identifying novel biomarkers are of paramount importance for improving patient outcomes. Prior research has demonstrated aberrant expression of Zinc Finger DHHC-Type Palmitoyltransferase 9 (ZDHHC9) in various malignancies, but its specific function and clinical significance in osteosarcoma remain poorly defined. This study employed a comprehensive approach integrating bioinformatic analyses and experimental validation to investigate the functional role and clinical relevance of ZDHHC9 in osteosarcoma. Bioinformatic analyses (TIMER, GEPIA, TCGA databases) were utilized to examine ZDHHC9 expression across a pan-cancer landscape and its association with survival in osteosarcoma patients. Immunohistochemistry (IHC) and Western blotting were employed to analyze ZDHHC9 expression in clinical osteosarcoma specimens and cell lines. In vitro assays (CCK-8, colony formation, wound healing, Transwell, flow cytometry, and Western blotting) were performed to evaluate the impact of ZDHHC9 on osteosarcoma cell proliferation, migration, and invasion. Proteomic sequencing, molecular docking, and co-immunoprecipitation (Co-IP) experiments were conducted to explore the interaction between ZDHHC9 and KRAS. Western blotting was used to analyze the regulation of Raf1, ERK1/2, and p-ERK1/2 expression by ZDHHC9 and KRAS. Finally, a xenograft model was employed to assess the effect of ZDHHC9 on in vivo tumor growth. Bioinformatic analyses revealed that elevated ZDHHC9 expression correlates with poor prognosis in osteosarcoma. IHC and Western blotting confirmed significantly increased ZDHHC9 expression in osteosarcoma tissues compared to adjacent non-tumorous tissues. Upregulation of ZDHHC9 was significantly associated with elevated Ki67 levels and advanced Enneking stage, suggesting a link to tumor aggressiveness. In vitro studies demonstrated that downregulation of ZDHHC9 suppressed osteosarcoma cell proliferation, migration, and invasion, while promoting apoptosis. Conversely, upregulation of ZDHHC9 elicited opposite effects. Proteomic sequencing revealed that ZDHHC9 knockdown significantly downregulated the expression of proteins associated with the RAS/MAPK signaling pathway. Functional experiments showed that overexpression of KRAS partially abrogated the inhibitory effects of ZDHHC9 knockdown on cell proliferation, migration, and invasion. Molecular docking and Co-IP experiments confirmed a specific interaction between ZDHHC9 and KRAS, leading to activation of the RAS/MAPK signaling pathway. ZDHHC9 promotes osteosarcoma cell proliferation, migration, and invasion by enhancing KRAS-mediated activity of the RAS/MAPK signaling pathway, which promotes cell cycle progression while inhibiting apoptosis. Data are available via ProteomeXchange with identifier PXD066322.

Background: Osteosarcoma (OS) patients face the challenge of having few effective therapeutic drugs. Solasonine(SS)is an active component of TCM against OS cells. This study aims to identify the key targets of solasonine in treating OS.Methods: In this study, the transcriptome data and related gene sets were first downloaded from public databases. Subsequently, candidate targets were obtained by intersecting differentially expressed genes (DEGs) with solasonine and OS disease targets. Key targets were then identified through regression analyses, and a prognostic model was constructed. A nomogram was subsequently constructed using the key targets. The functions and immune microenvironment, as well as the structure, regulatory network, and molecular docking of these key targets, were then analyzed. The expression level of the candidate targets in osteosarcoma cells was verified in RT-qPCR experiments, and the effect of solasonine on the malignant biological behavior of osteosarcoma cells was verified.Results: DEGs, targets corresponding to solasonine, and OS-related disease targets were intersected to obtain 37 candidate targets. Subsequent regression analyses identified 5 key targets (ATP1A1, CLK1, SIGMAR1, PYGM, HSP90B1). It was further demonstrated that the OS prognostic model constructed using these key targets was robust. The constructed nomogram provided an excellent predictive model. Moreover, some pathways, such as cytokine-cytokine receptor interaction, were significantly enriched, and there were 4 significantly different immune cells and 3 significantly different immune checkpoints (P<0.05). Additionally, natural killer cells and activated B cells were significantly positively correlated (cor = 0.68, P < 0.001). The subsequent regulatory network included transcription factors regulating the 5 targets. All key targets showed favorable molecular docking effects with SS. The target genes all exhibited higher expression in osteosarcoma cell lines(P<0.05). Solasonine can inhibit the malignant biological behavior of cell proliferation, migration and invasion.Conclusion: In this study, ATP1A1, CLK1, SIGMAR1, PYGM, and HSP90B1 were identified as key targets of solasonine in the treatment of OS, and they were found to have reference significance for the treatment of OS. SS can be a potential drug for the treatment of osteosarcoma.