Analysis of the value of potential biomarker S100-A8 protein in the diagnosis and pathogenesis of spinal tuberculosis.

Objectives: The objective of this study is to evaluate the value of S100-A8 protein as a diagnostic marker for spinal tuberculosis and to explore its role in the potential pathogenesis of spinal tuberculosis (STB). Methods: The peripheral blood of 100 spinal tuberculosis patients admitted to the General Hospital of Ningxia Medical University from September 2018 to June 2021 were collected as the observation group, and the peripheral blood of 30 healthy medical examiners were collected as the control group. Three samples from the observation group and three samples from the control group were selected for proteomics detection and screening of differential proteins. Kyoto Encyclopedia of Genes (KEGG) was used to enrich and analyze related signaling pathways to confirm the target protein. The serum expression levels of the target proteins were determined and compared between the two groups using enzyme-linked immunosorbent assay (ELISA). Statistical methods were used to evaluate the value of target protein as a diagnostic marker for STB. A macrophage model of Mycobacterium tuberculosis infection was constructed and S100-A8 small interfering RNA was used to investigate the molecular mechanism of the target protein. Results: S100-A8 protein has the value of diagnosing spinal tuberculosis (AUC = 0.931, p < 0.001), and the expression level in the peripheral blood of the observation group (59.04 ± 19.37 ng/mL) was significantly higher than that of the control group (43.16 ± 10.07 ng/mL) (p < 0.05). S100-A8 protein expression showed a significant positive correlation with both CRP and ESR values (p < 0.01). Its AUCs for combined bacteriological detection, T-SPOT results, diagnostic imaging, antacid staining results, and pathological results were 0.705 (p < 0.05), 0.754 (p < 0.01), 0.716 (p < 0.01), 0.656 (p < 0.05), and 0.681 (p < 0.01), respectively. Lack of S100-A8 leads to a significant decrease in the expression levels of TLR4 and IL-17A in infected macrophages. Conclusion: S100-A8 protein is differentially expressed in the peripheral blood of patients with spinal tuberculosis and healthy individuals and may be a novel candidate biomarker for the diagnosis of spinal tuberculosis. The feedback loop on the S100-A8-TLR4-IL-17A axis may play an important role in the inflammatory mechanism of spinal tuberculosis.

Plasma metabolomic analysis reveals the metabolic characteristics and potential diagnostic biomarkers of spinal tuberculosis.

Objectives: This study aimed to conduct a non-targeted metabolomic analysis of plasma from patients with spinal tuberculosis (STB) to systematically elucidate the metabolomic alterations associated with STB, and explore potential diagnostic biomarkers for STB. Methods: From January 2020 to January 2022, 30 patients with spinal tuberculosis (STBs) clinically diagnosed at the General Hospital of Ningxia Medical University and 30 age- and sex-matched healthy controls (HCs) were selected for this study. Using ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-QTOF/MS) based metabolomics, we analyzed the metabolic profiles of 60 plasma samples. Statistical analyses, pathway enrichment, and receiver operating characteristic (ROC) analyses were performed to screen and evaluate potential diagnostic biomarkers. Results: Metabolomic profiling revealed distinct alterations between the STBs and HCs cohorts. A total of 1635 differential metabolites were screened, functionally clustered, and annotated. The results showed that the differential metabolites were enriched in sphingolipid metabolism, tuberculosis, cutin, suberine and wax biosynthesis, beta-alanine metabolism, methane metabolism, and other pathways. Through the random forest algorithm, LysoPE (18:1(11Z)/0:0), 8-Demethyl-8-formylriboflavin 5'-phosphate, Glutaminyl-Gamma-glutamate, (2R)-O-Phospho-3-sulfolactate, and LysoPE (P-16:0/0:0) were determined to have high independent diagnostic value. Conclusions: STBs exhibited significantly altered metabolite profiles compared with HCs. Here, we provide a global metabolomic profile and identify potential diagnostic biomarkers of STB. Five potential independent diagnostic biomarkers with high diagnostic value were screened. This study provides novel insights into the pathogenesis, diagnosis, and treatment strategies of STB.

Integrative analysis of the expression profiles of whole coding and non-coding RNA transcriptomes and construction of the competing endogenous RNA networks for chronic obstructive pulmonary disease.

The pathogenesis of Chronic Obstructive Pulmonary Disease (COPD) is implicated in airway inflammation, oxidative stress, protease/anti-protease and emphysema. Abnormally expressed non-coding RNAs (ncRNAs) play a vital role in regulation of COPD occurrence and progression. The regulatory mechanisms of the circRNA/lncRNA-miRNA-mRNA (competing endogenous RNA, ceRNA) networks might facilitate our cognition of RNA interactions in COPD. This study aimed to identified novel RNA transcripts and constructed the potential ceRNA networks of COPD patients. Total transcriptome sequencing of the tissues from patients with COPD (COPD) (n = 7) and non-COPD control subjects (Normal) (n = 6) was performed, and the expression profiles of differentially expressed genes (DEGs), including mRNAs, lncRNAs, circRNAs, and miRNAs, were analyzed. The ceRNA network was established based on the miRcode and miRanda databases. Kyoto Encyclopedia of Genes and Genomes (KEGG), Gene Ontology (GO), Gene Set Enrichment Analysis (GSEA), and Gene set variation analysis (GSVA) were implemented for functional enrichment analysis of DEGs. Finally, CIBERSORTx was extracted to analyze the relevance between hub genes and various immune cells.The Starbase and JASPAR databases were used to construct hub-RNA binding proteins (RBPs) and lncRNA-transcription factor (TF) interaction networks. A total of 1,796 mRNAs, 2,207 lncRNAs, and 11 miRNAs showed differentially expression between the lung tissue samples from the normal and COPD groups. Based on these DEGs, lncRNA/circRNA-miRNA-mRNA ceRNA networks were constructed respectively. In addition, ten hub genes were identified. Among them, RPS11, RPL32, RPL5, and RPL27A were associated with the proliferation, differentiation, and apoptosis of the lung tissue. The biological function revealed that TNF-α via NF-kB and IL6/JAK/STAT3 signaling pathways were involved in COPD. Our research constructed the lncRNA/circRNA-miRNA-mRNA ceRNA networks, filtrated ten hub genes may regulate the TNF-α/NF-κB, IL6/JAK/STAT3 signally pathways, which indirectly elucidated the post-transcriptional regulation mechanism of COPD and lay the foundation for excavating the novel targets of diagnosis and treatment in COPD.

Preparation, characterization, and in-vitro cytotoxicity of nanoliposomes loaded with anti-tubercular drugs and TGF-ß1 siRNA for improving spinal tuberculosis therapy.

BACKGROUND: Tuberculosis (TB) represents a bacterial infection affecting many individuals each year and potentially leading to death. Overexpression of transforming growth factor (TGF)-ß1 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-ß1-specific short interfering RNA (siRNA) delivery system employing nanoliposomes. METHODS: In the current study, siTGF-ß1 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-ß1 siRNA. RESULTS: HRZ/siTGF-ß1 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-ß1 could significantly downregulate TGF-ß1 in THP-1-derived human macrophages in vitro. CONCLUSION: These findings suggested that HRZ-loaded nanoliposomes with siTGF-ß1 have the potential for improving spinal tuberculosis chemotherapy via nano-encapsulation of anti-TB drugs.

Analysis of the Value of Serum Biomarker LBP in the Diagnosis of Spinal Tuberculosis.

Objective: To investigate the correlation between the expression of lipopolysaccharide-binding protein (LBP) in peripheral blood of spinal tuberculosis and clinical diagnosis and to evaluate its value as a diagnostic marker of spinal tuberculosis. Methods: In the experimental group, clinical history data and peripheral blood were collected from 100 patients with spinal tuberculosis who were admitted to the Department of Spine Surgery, General Hospital of Ningxia Medical University from May 2017 to May 2020, and peripheral blood was collected from 30 healthy volunteers in the control group. Screening of differential LBP expression by proteomics and ELISA to verify its expression in peripheral blood of spinal tuberculosis patients. t-test, Spearman analysis, linear regression and ROC curve were used to evaluate the diagnostic value of LBP in peripheral blood for spinal tuberculosis. Results: The expression of LBP protein in peripheral blood is significantly higher in patients with spinal tuberculosis than in the normal population; LBP assay values were significantly and positively correlated with CRP and ESR values (P < 0.01); the AUC of LBP in the diagnosis of spinal tuberculosis for pathological examination, bacteriological culture, T-cell spot test for tuberculosis infection (T-SPOT), imaging diagnosis, and acid fast bacillus were, respectively, 0.677 (P < 0.01), 0.707 (P < 0.01), 0.751 (P < 0.01), 0.714 (P < 0.01), and 0.656 (P < 0.05), and there was a correlation between LBP and the diagnostic evaluation of spinal tuberculosis. Conclusion: LBP could be a new candidate biomarker for the diagnosis of spinal tuberculosis.

Pathological features of Brucella spondylitis: A single-center study.

OBJECTIVE: To explore the pathological features of Brucella spondylitis (BS) under the optical microscope, thus providing pathological references for the diagnosis. METHODS: We retrospectively analyzed 70 BS patients (42 males and 28 females, mean age 52.01 ± 10.77 [20-74] years) admitted in the Department of Spine Surgery, the General Hospital of Ningxia Medical University, from January 2013 to December 2020. Their medical history, clinical manifestations, laboratory test results, imaging findings and bacteriological culture results were collected. Among them, 5, 5, 43, 4 and 13 cases demonstrated involvement into the cervical vertebra, thoracic vertebra, lumbar vertebra, thoracolumbar vertebra and lumbosacral vertebra, respectively. Notably, L4 showed pathology in 32 cases. Pathological features of BS were analyzed by H&E staining of granulation tissues, sclerotic bones, sequestrums, and intervertebral discs. RESULTS: 42 cases underwent bacterial culture, of which 4 were positive, and the positive rate of bacterial culture was only 9.5%. The highest Vas score was 7, the lowest was 4, and the average was 5.76 ± 0.89. The highest CRP was 153 mg/L, the lowest was 0.98 mg/L, and the average was 30.98 ± 33.79 mg/L. The highest ESR is 112 mm/h, the lowest is 5 mm/h, and the average is 49.34 ± 27.73 mm/h. Under the optical microscope, BS manifested acute or chronic inflammation. Acute inflammatory features included neutrophil and eosinophil infiltration, necrosis, and abscesses, while chronic inflammatory features included lymphocyte, plasma cell, fibrous tissue and monocyte infiltration, hyaline degeneration, angiogenesis and hyperplasia of other tissues. Other features included vasodilation, hemorrhage, granulomas and multinucleated giant cell infiltration. In the present study, chronic inflammation was observed in 25 cases, in-acute-phase chronic inflammation in 45 cases, and acute inflammation in no cases. Pathological features of BS under the microscope included foam cell reaction in 46 cases, histiocytic reaction in 24 cases and eosinophilic abscesses in 6 cases. Eosinophil infiltration was observed in 45 cases (mainly during the acute phase of chronic inflammation) and massive eosinophilic abscesses in 6 cases. Granulation tissue hyperplasia followed inflammatory repair in 25 BS cases, and was generally boosted in the acute phase of chronic inflammation. Multinucleated giant cell infiltration and granulomas were less observed in BS cases, which differed from pathological features of spinal tuberculosis. CONCLUSIONS: Chronic inflammation or in-acute-phase chronic inflammation is the main pathological feature of BS, while the single acute inflammation is less observed in BS cases. Foam cell reaction and histiocytic reaction scale up during the acute phase of chronic inflammation, and some BS patients may develop massive eosinophilic abscesses. Granulation tissue hyperplasia, rather than multinucleated giant cell infiltration and granulomas, serve as pathological reference for the differential diagnosis of BS and spinal tuberculosis.