Background: Tuberculosis (TB) represents a bacterial infection affecting many individuals each year and potentially leading to death. Overexpression of transforming growth factor (TGF)-β1 has a primary immunomodulatory function in human tuberculosis. This work aimed to develop nanoliposomes to facilitate the delivery of anti-tubercular products to THP-1-derived human macrophages as Mycobacterium host cells and to evaluate drug efficiencies as well as the effects of a TGF-β1-specific short interfering RNA (siRNA) delivery system employing nanoliposomes. Methods: In the current study, siTGF-β1 nanoliposomes loaded with the anti-TB drugs HRZ (isoniazid, rifampicin, and pyrazinamide) were prepared and characterized in vitro, determining the size, zeta potential, morphology, drug encapsulation efficiency (EE), cytotoxicity, and gene silencing efficiency of TGF-β1 siRNA. Results: HRZ/siTGF-β1 nanoliposomes appeared as smooth spheres showing the size and positive zeta potential of 168.135 ± 0.5444 nm and + 4.03 ± 1.32 mV, respectively. Drug EEs were 90%, 88%, and 37% for INH, RIF, and PZA, respectively. Meanwhile, the nanoliposomes were weakly cytotoxic towards human macrophages as assessed by the MTT assay. Nanoliposomal siTGF-β1 could significantly downregulate TGF-β1 in THP-1-derived human macrophages in vitro. Conclusion: These findings suggested that HRZ-loaded nanoliposomes with siTGF-β1 have the potential for improving spinal tuberculosis chemotherapy via nano-encapsulation of anti-TB drugs.

Spinal Tuberculosis (STB) constitutes a common form of tuberculosis and is more widespread in developing countries. Not only is it one of the principal causes of spinal deformity but also it may trigger severe neurological impairments, thereby significantly influencing the quality of life of patients. Additionally, microRNAs (miRNAs) participate in the signal transduction among macrophages through multiple means. In the earlier work of our research group, spinal tuberculosis tissues were subjected to miRNA differential analysis and sequencing. The results indicated that miR-29a-3p was under-expressed in patients with spinal tuberculosis relative to the normal population. Nevertheless, it remains unknown at present how methyltransferase 14(METTL14)-mediated miR-29a-3p regulates the inflammatory response of spinal tuberculosis via the Mitogen-Activated Protein Kinase Kinase 6(MAP2K6) signaling pathway. This study aimed to validate the expression levels of METTL14 and miR-29a-3p in spinal tuberculosis and explore the regulatory role of METTL14 in mediating miR-29a-3p via the MAP2K6 signaling pathway during spinal tuberculosis-associated inflammation. Twenty cases of peripheral blood samples from patients with spinal tuberculosis and normal individuals were collected respectively. Through the construction of a BCG-infected THP-1 macrophage model, the expression of METTL14 and miR-29a-3p in spinal tuberculosis was verified by means of clinical sample analysis and cell experiments such as RT-qPCR, immunohistochemistry, and Western blot. The regulation of METTL14-mediated miR-29a-3p through the MAP2K6 signaling pathway in the inflammatory response of spinal tuberculosis was explored via immunofluorescence, RT-qPCR, and ELISA. The expressions of METTL14 and miR-29a-3p were significantly downregulated in patients with spinal tuberculosis, while the expression of MAP2K6 was upregulated. METTL14 regulated the expression of miR-29a-3p through m6A modification, thereby targeting and inhibiting the expression of MAP2K6. Overexpression of METTL14 and miR-29a-3p could suppress the MAP2K6 signaling pathway, alleviated inflammatory responses and spinal tissue damage; conversely, inhibition of METTL14 and miR-29a-3p activated the MAP2K6 signaling pathway, intensifying inflammatory responses and spinal tissue damage. This research uncovers the crucial role of the METTL14/ miR-29a-3p/ MAP2K6 axis in spinal tuberculosis, providing experimental evidence for the utilization of non-coding RNA as molecular targets in spinal tuberculosis, providing references for the study of the pathogenesis of spinal tuberculosis, and offering novel ideas and perspectives for the diagnosis and treatment of spinal tuberculosis. This indicates that the treatment targeting METTL14 may provide a new strategy for the treatment of STB and facilitate its early clinical detection.

Rationale: Highly virulent multidrug-resistant Klebsiella pneumoniae (KP) is becoming more and more common in clinical practice, especially the rise of carbapenem-resistant KP in clinical practice, resulting in the emergence of KP liver abscess in Ningxia, China. For the prognosis of liver abscess patients, it is particularly important to identify the types of pathogens and identify antibiotics that are sensitive to the pathogens. Patient concerns: A 73-year-old man from China presents to our hospital with abdominal pain, jaundice and fever. Patients have no obvious cause of abdominal pain, abdominal distension, and abdominal pain is persistent. Abdominal examination showed hepatomegaly, no tenderness 2 cm from the right costal margin, abdominal distension and other general examinations did not have obvious abnormalities. He had no history of hypertension and diabetes, ERCP was performed for cholangiocarcinoma 1 year before the current visit, and no significant complications occurred. Diagnoses: His initial diagnosis was obstructive cholangitis, and computed tomographic images and liver drainage fluid bacterial culture and genetic polymerase chain reaction tests later determined that the patient had KP liver abscess. Interventions: Drainage by liver catheter and antibiotic treatment for 7 weeks. Outcomes: The patient liver abscess is basically gone. Lession: It is particularly important to optimize the diagnosis of liver abscess pathogens for timely and effective treatment of patients.

Background: The association between peripheral blood immune-inflammatory markers and neurological involvement in patients with spinal tuberculosis, and to develop a predictive model for early risk assessment. Spinal tuberculosis with neurological impairment is a major cause of disability, yet simple, cost-effective early warning indicators remain lacking. Peripheral blood immune-inflammatory markers are valuable in infectious disease prognostication, but their predictive role in spinal tuberculosis neurological involvement remains unclear. Methods: A retrospective study was conducted on 294 patients with spinal tuberculosis who underwent surgical treatment in the Department of Orthopedics, General Hospital of Ningxia Medical University, between December 2019 and December 2024. Patients were stratified into two groups: the uncomplicated spinal tuberculosis group (n=194, ASIA grade E) and the neurological involvement group (n=100, ASIA grades A–D). Fasting venous blood test results collected at the initial presentation were analyzed. Immune-inflammatory markers, including lymphocyte percentage (LYM), mixed cell percentage (MXD), and platelet-to-lymphocyte ratio (PLR), were compared between the two groups. Univariate and multivariate logistic regression analyses were performed to identify influencing factors, followed by the establishment of a predictive model. Receiver operating characteristic (ROC) curves and a nomogram were constructed to evaluate the diagnostic efficacy of the model. Results: The neurological involvement group exhibited a significantly lower LYM [23.150% (16.900, 29.525) vs. 26.150% (19.750, 32.900)], and significantly higher MXD [9.000% (7.375, 10.200) vs. 8.100% (6.725, 9.900)] and PLR compared with the uncomplicated group ( P < 0.05). Multivariate analysis revealed that LYM (odds ratio [OR]=0.961, 95% confidence interval [CI]: 0.935–0.989) and MXD (OR = 1.107, 95% CI: 1.013–1.209) were independent predictors of neurological involvement in spinal tuberculosis. Specifically, each 1% increase in LYM was associated with a 3.9% reduction in the risk of neurological impairment, whereas each 1% increase in MXD correlated with a 10.7% increase in risk. The combined predictive model achieved an AUC of 0.803 (95% CI: 0.749–0.857), with a sensitivity of 70.0% and specificity of 80.9%. The calibration curve confirmed good model fit (χ²=4.215, P = 0.837). In addition, the Brier score was 0.185, indicating favorable overall accuracy of the probabilistic predictions. Conclusion: Decreased peripheral blood LYM and increased MXD are independent risk factors for neurological involvement in spinal tuberculosis. The combined predictive model shows favorable diagnostic efficacy and calibration. The nomogram is a simple, economical clinical tool for early high-risk patient identification, guiding individualized treatment decisions.

Objective: To validate the safety and multicenter technical feasibility of an innovatively designed percutaneous paramedian small-angle lateral intervertebral foramen Kambin’s triangle approach for lumbar puncture biopsy in early spinal infection (SI), and to evaluate the diagnostic yield and etiological identification rate of the tNGS-based multimodal etiological detection protocol using specimens obtained via this approach. Methods: This was a multicenter retrospective cohort study that enrolled 94 consecutive patients with SI who underwent the aforementioned puncture biopsy at 3 medical centers between January 2024 and January 2026. We systematically evaluated the procedure-related indicators and perioperative complications of this puncture approach. All puncture specimens were synchronously subjected to histopathological examination, routine bacterial culture, Xpert MTB/RIF assay, and tNGS detection. With the final comprehensive clinical diagnosis as the reference standard, we compared and analyzed the diagnostic performance of each detection method. Results: All 94 patients successfully completed biopsy via this modified puncture approach, with an overall puncture-related complication rate of only 7.45%. All adverse events were mild, and no severe complications occurred, and the approach was feasible and appeared safe in this retrospective cohort, though prospective validation across multiple operators is warranted before widespread clinical adoption. For etiological detection, the overall positive rate of tNGS reached 87.23%, which was significantly higher than that of routine bacterial culture (25.53%), specific histopathological diagnosis (17.02%), and Xpert MTB/RIF assay (17.02%) (all P < 0.0001). Meanwhile, tNGS maintained a stable and high detection rate across all major SI subtypes. The tNGS-centered multimodal combined detection achieved an overall positive rate of 91.49%. Notably, in early-stage cases where histopathology only indicated non-specific inflammatory cell infiltration, the combined detection achieved an etiology confirmation rate of 95.65%, effectively compensating for the deficiency of detection methods in early diagnosis. Conclusions: The puncture approach designed in this study addresses the core bottleneck of specimen acquisition in early SI. The tNGS-based multimodal detection system using this approach has excellent diagnostic performance in early-stage infection, and can provide critical evidence-based support for the early accurate diagnosis and treatment as well as targeted antimicrobial therapy of SI.

Background: The safety and efficacy of placing pedicle screws within infected vertebral bodies during surgery for acute pyogenic spondylitis remains controversial. The conventional “skip-level” fixation strategy avoids instrumentation at the infected site but may compromise spinal biomechanical stability and alignment. Conversely, fixation that includes the infected vertebrae can provide immediate stability, facilitate deformity correction, and promote bone fusion, although it carries potential risks of infection recurrence and implant failure. Objective: To describe the short-term outcomes of direct pedicle screw fixation in infected vertebrae for acute pyogenic spondylitis, combined with radical debridement and targeted antimicrobial therapy. Methods: A retrospective analysis was conducted on 32 patients with pyogenic spondylitis who underwent surgical treatment between January 2021 and January 2025. All patients had a confirmed etiological diagnosis obtained via microbiological culture or next-generation sequencing (NGS) of surgical specimens and received individualized perioperative anti-infective therapy guided by clinical pharmacists. The surgical protocol included debridement of the infectious focus and posterior pedicle screw fixation incorporating the infected vertebrae, performed via single-stage posterior, single-stage combined anterior-posterior, or staged combined anterior-posterior approaches. Primary outcomes focused on perioperative adverse events, including implant-related complications, infection recurrence, and wound healing. Secondary outcomes assessed clinical efficacy and involved comparisons of Visual Analog Scale (VAS) pain scores, Oswestry Disability Index (ODI), inflammatory markers (erythrocyte sedimentation rate [ESR], C-reactive protein [CRP], and white blood cell count [WBC]), spinal Cobb angle, Frankel neurological grade, and bone fusion rates. Inflammatory markers and Cobb angle were assessed preoperatively and at 1, 3, and 6 months postoperatively; VAS and ODI were assessed preoperatively and at 1, 3, 6, and 12 months postoperatively; and bone fusion was assessed at 6 and 12 months postoperatively. Bone fusion was independently evaluated by two blinded assessors using Bridwell or modified Lenke criteria. Results: All 32 patients completed the 12-month follow-up. One osteoporotic patient developed asymptomatic minor screw loosening, and one diabetic patient experienced delayed wound healing that resolved after treatment. No implant failure, infection recurrence, or severe neurological complications occurred. Friedman tests demonstrated significant overall time effects for all dynamic indicators (all P < 0.0001): ESR ( χ² (3) = 62.87, W = 0.65), CRP ( χ² (3) = 67.22, W = 0.70), WBC ( χ² (3) = 37.93, W = 0.40), Cobb angle ( χ² (3) = 34.71, W = 0.36), VAS ( χ² (4) = 95.90, W = 0.75), and ODI ( χ² (4) = 127.02, W = 0.99). By 6 months, inflammatory markers had decreased markedly: ESR from 55.19 ± 31.05 to 6.00 (3.50, 12.25) mm/h; CRP from 34.10 (13.97, 85.15) to 2.12 (0.96, 3.90) mg/L; and WBC from 9.08 (5.75, 11.27) to 5.46 ± 1.01 ×10 ⁹ /L. The Cobb angle improved from 7.94 (6.84, 14.22)° to 8.34 ± 2.43°. At 12 months, VAS improved from 4.00 (4.00, 5.00) to 1.00 (0.75, 1.00), and ODI decreased from 28.72 ± 8.82% to 5.00 (2.25, 8.00)%. Neurological function recovered to Frankel grade E in 93.8% (30/32) of patients. The overall descriptive fusion rate (successful fusion in at least one graft site) was 78.13% at 6 months and 93.75% at 12 months postoperatively. Conclusions: In this single-arm retrospective study of 32 patients with acute pyogenic spondylitis, direct pedicle screw fixation involving infected vertebrae was performed in conjunction with radical debridement and targeted antimicrobial therapy. At 12 months, patients demonstrated favorable short-term outcomes, including a low incidence of implant-related complications, significant improvements in pain and disability, a high radiographic fusion rate, and no observed infection recurrence. These findings provide preliminary evidence that this comprehensive protocol is a viable strategy for achieving favorable short-term outcomes in selected patients, and may serve as a practical reference for surgeons considering direct instrumentation in infected vertebrae.