CD_99 G1 neutrophils modulate osteogenic differentiation of mesenchymal stem cells in the pathological process of ankylosing spondylitis.

OBJECTIVES: This study aimed to identify the types and heterogeneity of cells within the spinal enthesis and investigate the underlying mechanisms of osteogenesis. METHODS: Single-cell RNA sequencing was used to identify cell populations and their gene signatures in the spinal enthesis of five patients with ankylosing spondylitis (AS) and three healthy individuals. The transcriptomes of 40 065 single cells were profiled and divided into 7 clusters: neutrophils, monocytic cells, granulomonocytic progenitor_erythroblasts, T cells, B cells, plasma cells and stromal cells. Real-time quantitative PCR, immunofluorescence, flow cytometry, osteogenesis induction, alizarin red staining, immunohistochemistry, short hairpin RNA and H&E staining were applied to validate the bioinformatics analysis. RESULTS: Pseudo-time analysis showed two differentiation directions of stromal cells from the mesenchymal stem cell subpopulation MSC-C2 to two Cxcl12-abundant-reticular (CAR) cell subsets, Osteo-CAR and Adipo-CAR, within which three transcription factors, C-JUN, C-FOS and CAVIN1, were highly expressed in AS and regulated the osteogenesis of mesenchymal stem cells. A novel subcluster of early-stage neutrophils, CD99_G1, was elevated in AS. The proinflammatory characteristics of monocyte dendritic cell progenitor-recombinant adiponectin receptor 2 monocytic cells were explored. Interactions between Adipo-CAR cells, CD99_G1 neutrophils and other cell types were mapped by identifying ligand-receptor pairs, revealing the recruitment characteristics of CD99_G1 neutrophils by Adipo-CAR cells and the pathogenesis of osteogenesis induced in AS. CONCLUSIONS: Our results revealed the dynamics of cell subpopulations, gene expression and intercellular interactions during AS pathogenesis. These findings provide new insights into the cellular and molecular mechanisms of osteogenesis and will benefit the development of novel therapeutic strategies.

CX3CL1 promotes M1 macrophage polarization and osteoclast differentiation through NF-κB signaling pathway in ankylosing spondylitis in vitro.

BACKGROUND: Ankylosing spondylitis (AS) is an autoimmune disease with a genetic correlation and is characterized by inflammation in the axial skeleton and sacroiliac joints. Many AS patients also have inflammatory bowel diseases (IBD), but the underlying causes of intestinal inflammation and osteoporosis in AS are not well understood. CX3CL1, a protein involved in inflammation, has been found to be up-regulated in AS patients and AS-model mice. METHODS: The authors investigated the effects of CX3CL1 on AS by studying its impact on macrophage polarization, inflammation factors, and osteoclast differentiation. Furthermore, the effects of inhibiting the NF-κB pathway and blocking CX3CL1 were assessed using BAY-117082 and anti-CX3CL1 mAb, respectively. AS model mice were used to evaluate the effects of anti-CX3CL1 mAb on limb thickness, spine rupture, and intestinal tissue damage. RESULTS: The authors found that CX3CL1 increased the expression of M1-type macrophage markers and inflammation factors, and promoted osteoclast differentiation. This effect was mediated through the NF-κB signaling pathway. Inhibition of the NF-κB pathway prevented M1-type macrophage polarization, reduced inflammation levels, and inhibited osteoclast differentiation. Injection of anti-CX3CL1 mAb alleviated limb thickness, spine rupture, and intestinal tissue damage in AS model mice by inhibiting M1-type macrophage polarization and reducing intestinal tissue inflammation. CONCLUSIONS: The study demonstrated that up-regulated CX3CL1 promotes M1-type macrophage polarization and osteoclast differentiation through the NF-κB signaling pathway. Inhibition of this pathway and blocking CX3CL1 can alleviate inflammation and bone destruction in AS. These findings contribute to a better understanding of the pathogenesis of AS and provide a basis for clinical diagnosis and treatment.

Impact of immune regulation and differentiation dysfunction of mesenchymal stem cells on the disease process in ankylosing spondylitis and prospective analysis of stem cell transplantation therapy.

Ankylosing spondylitis (AS) is a rheumatic bone and joint disease caused by inflammation, erosion, and pathological bone formation. The pathological features of chronic inflammation, bone destruction, and pathological ossification occur due to the disruption of the body's immune regulation and altered bone remodeling balance. Mesenchymal stem cells (MSCs) have multidirectional differentiation potential and immunomodulatory functions and play an important role in immune regulation and bone formation. The immune regulation and osteogenic capacity of MSCs in AS are altered by factors such as genetic background, internal environment, infection, and mechanical forces that drive disease development. This review further evaluates the role of MSCs dysfunction in inflammation and pathological bone formation by analyzing the effects of the above-mentioned factors on MSCs function and also looks forward to the prospects of MSCs in treating AS, providing some ideas for an in-depth study of inflammation and ectopic ossification.

BZR1 and BES1 participate in regulation of glucosinolate biosynthesis by brassinosteroids in Arabidopsis.

The effect of 24-epibrassinolide (EBR) on glucosinolate biosynthesis in Arabidopsis thaliana was investigated in the present study by using mutants and transgenic plants involved in brassinosteroid (BR) biosynthesis and signal transduction, as well as glucosinolate biosynthesis. The results showed that EBR significantly decreased the contents of major aliphatic glucosinolates including glucoiberin (S3), glucoraphanin (S4), and glucoerucin (T4), as well as the indolic glucosinolates glucobrassicin (IM) and neoglucobrassicin (1IM). In addition, a significantly higher level of glucosinolates accumulated in the BR-deficient mutant cpd and a dramatically lower glucosinolate content in the transgenic plant DWF4-ox overexpressing the BR biosynthetic gene DWF4 compared with their related wild-types, confirmed the repressing effect of BR on glucosinolate biosynthesis. BRI1, the receptor of BR signal transduction, was involved in regulation of glucosinolate biosynthesis by BR. Furthermore, the observation of reduced content of glucosinolates and lower expression levels of glucosinolate biosynthetic genes in 35S-BZR1/bzr1-1D and bes1-D plants compared with the corresponding wild-types suggested that BZR1 and BES1, two important components in BR signal transduction, are responsible for the inhibiting role of BR in glucosinolate biosynthesis. The disappearance of the repressing effect of BR on glucosinolate content in the myb28, myb34, and myb122 mutants indicated that these three MYB factors are important for the regulation of BR in glucosinolate biosynthesis.

Recent Advances in Halogen-Free Flame Retardants for Polyolefin Cable Sheath Materials.

With the continuous advancements of urbanization, the demand for power cables is increasing to replace overhead lines for energy transmission and distribution. Due to undesirable scenarios, e.g., the short circuit or poor contact, the cables can cause fire. The cable sheath has a significant effect on fire expansion. Thus, it is of great significance to carry out research on flame-retardant modification for cable sheath material to prevent fire accidents. With the continuous environmental concern, polyolefin (PO) is expected to gradually replace polyvinyl chloride (PVC) for cable sheath material. Moreover, the halogen-free flame retardants (FRs), which are the focus of this paper, will replace the ones with halogen gradually. The halogen-free FRs used in PO cable sheath material can be divided into inorganic flame retardant, organic flame retardant, and intumescent flame retardant (IFR). However, most FRs will cause severe damage to the mechanical properties of the PO cable sheath material, mainly reflected in the elongation at break and tensile strength. Therefore, the cooperative modification of PO materials for flame retardancy and mechanical properties has become a research hotspot. For this review, about 240 works from the literature related to FRs used in PO materials were investigated. It is shown that the simultaneous improvement for flame retardancy and mechanical properties mainly focuses on surface treatment technology, nanotechnology, and the cooperative effect of multiple FRs. The principle is mainly to improve the compatibility of FRs with PO polymers and/or increase the efficiency of FRs.

Dehydroquinate Synthase Directly Binds to Streptomycin and Regulates Susceptibility of Mycobacterium bovis to Streptomycin in a Non-canonical Mode.

Antimicrobial resistance (AMR) represents one of the main challenges in Tuberculosis (TB) treatment. Investigating the genes involved in AMR and the underlying mechanisms holds promise for developing alternative treatment strategies. The results indicate that dehydroquinate synthase (DHQS) regulates the susceptibility of Mycobacterium bovis BCG to first-line anti-TB drug streptomycin. Perturbation of the expression of aroB encoding DHQS affects the susceptibility of M. bovis BCG to streptomycin. Purified DHQS impairs in vitro antibacterial activity of streptomycin, but did not hydrolyze or modify streptomycin. DHQS directly binds to streptomycin while retaining its own catalytic activity. Computationally modeled structure analysis of DHQS-streptomycin complex reveals that DHQS binds to streptomycin without disturbing native substrate binding. In addition, streptomycin treatment significantly induces the expression of DHQS, thus resulting in DHQS-mediated susceptibility. Our findings uncover the additional function of DHQS in AMR and provide an insight into a non-canonical resistance mechanism by which protein hijacks antibiotic to reduce the interaction between antibiotic and its target with normal protein function retained.

Ultra-high synergetic intensity for humic acid removal by coupling bubble discharge with activated carbon.

Humic acid (HA) removal research focuses on the global water treatment industry. In this work, efficient HA degradation with an ultra-high synergetic intensity is achieved by combined bubble discharge with activated carbon (AC). Adding AC to the discharge greatly improves HA removal efficiency and degradation speed; the synergetic intensity reaches 651.52% in the combined system, and the adsorption residual on AC is 4.52%. After 90 min of treatment, the HA removal efficiency reaches 98.90%, 31.29%, and 7.61% in the plasma-AC combined, solo bubble discharge, and solo AC adsorption systems, respectively. During the plasma process, the number of pore structures and active sites and the amount of oxygen-containing functional groups on the AC surface increase, resulting in a higher adsorption capacity to reactive species (H2O2 and O3) and HA and promoting interactions on the AC surface. For HA mineralization, the presence of AC greatly promotes the destruction of aromatic structures and chromophoric HA functional groups.

Elevated serum granzyme B levels are associated with disease activity and joint damage in patients with rheumatoid arthritis.

OBJECTIVES: Little is known about the roles of granzyme B in rheumatoid arthritis (RA). We aimed to evaluate the serum level of granzyme B in patients with RA and determine relationships with clinical features and joint destruction of RA. METHODS: We enrolled 100 patients with RA, 50 patients with osteoarthritis (OA), and 50 healthy controls (HC). Granzyme B serum concentrations were measured by ELISA; we then analyzed associations between granzyme B levels, clinical features, and joint destruction by calculating Sharp scores and disease activity as measured by Disease Activity Score-28 based on erythrocyte sedimentation rate (DAS28-ESR) in patients with RA. RESULTS: Compared with HC and patients with OA, serum granzyme B levels in patients with RA were remarkably elevated. Serum granzyme B levels did not differ between patients with OA and HC. Granzyme B levels correlated with ESR, rheumatoid factor, swollen joint counts, joint erosion scores, total Sharp scores, and DAS28-ESR. Moreover, patients with RA with high disease activity had higher granzyme B levels. CONCLUSIONS: Serum granzyme B levels were elevated significantly in patients with RA and correlated positively with disease activity and joint destruction. Serum granzyme B may have potential applications in laboratory evaluation of patients with RA.

Effect of vertical cut on coronal coverage and rotation of tibial component in Oxford unicompartmental knee arthroplasty.

OBJECTIVE: This study was performed to investigate the influence of a standard Oxford vertical cut on the coronal coverage and rotation of the tibial component and determine whether a relationship exists between coverage and rotation. METHODS: We retrospectively analyzed 71 patients with anteromedial osteoarthritis of the knee treated by Oxford unicompartmental knee arthroplasty in one center from October 2016 to October 2017. The distance of coronal coverage was measured on a postoperative anteroposterior view of the tibial component. Two different reference lines between the lateral wall of the tibial component were defined as rotation angle α and ß, respectively, on a computed tomography scan. RESULTS: The mean distance was 0.3 ± 1.1 mm. The mean angle α and ß were 5.7° ± 4.6° and 8.4° ± 4.6°, respectively. There were no significant differences in the distance according to the tibial component rotation or in the α and ß angles according to the coronal coverage. No significant correlation was found between the α and ß angles and the distance. CONCLUSION: A standard tibial vertical cut caused various changes in coronal coverage and rotation of the tibial component. The rotation of the tibial component did not affect coverage within a certain range.

Inhibition of HIF-1α restrains fracture healing via regulation of autophagy in a rat model.

It has been demonstrated that bone fracture is associated with the activation of autophagy, and upregulation of autophagy could promote fracture healing. Previous study by our group demonstrated that activating the HIF-1α pathway via administration of cobalt (II) chloride (CoCl2) could promote fracture healing in vivo. However, the role of hypoxia-inducible factor-1α (HIF-1α) in autophagy remains unknown. In the current study, rats were divided into two groups following tibial fracture and treated with echinomycin or dimethyl sulfoxide (DMSO). Rats were sacrificed at 7, 14, 28 and 42 days after fracture. The evaluation of fracture healing was performed by micro-computed tomography. In addition, the effects of echinomycin on microtubule-associated protein 1 light chain 3 (LC3 II), runt-related transcription factor 2 (Runx2), alkaline phosphatase (ALP), Unc-51-like autophagy activating kinase 1 (ULK1) and P62 were detected at the mRNA and protein levels by reverse transcription-quantitative polymerase chain reaction, western blotting and immunohistochemistry. The results demonstrated that the expression of LC3 II was markedly decreased following systemic administration of echinomycin (0.05 mg/kg every other day for 42 days, intraperitoneally). Furthermore, the levels of Runx2, ALP and ULK1 were decreased, while those of P62 were increased, at the mRNA and protein levels in rats treated with echinomycin in vivo. In summary, the current study suggested that HIF-1α may serve an important role in fracture healing via the downregulation of autophagy.

Comparison of remote ischemic preconditioning and intermittent hypoxia training in fracture healing.

Fracture healing in elderly patients is an emerging public health concern. As non­drug treatments, intermittent hypoxia training (IHT) and remote ischemic preconditioning (RIPC) are considered to have substantial advantages and to aid fracture healing in elderly patients. The purpose of the present study was to evaluate and compare the effects of IHT and RIPC on fracture healing. Micro­computed tomography (micro­CT) and biomechanical testing were used to assess the morphology and structural properties of bone callus dissected from aged rats with tibial fractures. In addition, hypoxia­inducible factor­1α (HIF­1α) and its target gene, associated with the healing process, were investigated by reverse transcription­quantitative polymerase chain reaction and western blot analyses. The micro­CT­based parameters, including bone mineral density and trabecular number, were measured, and significant differences were identified between the experimental and control groups. The IHT group exhibited superior bone formation and mineralization rates compared with the RIPC group. The biomechanical testing revealed that the ultimate loading and stiffness values were significantly higher in the IHT group compared with those in the RIPC group. In accordance with previous studies, RIPC exerted a similar effect in increasing the expression of HIF­1α, and its downstream genes, throughout the course of healing. In addition, the IHT group exhibited increased expression levels of HIF­1α compared with the RIPC group. Taken together, the results suggested that IHT and RIPC significantly enhanced fracture healing; however, IHT exhibited superior bone formation and healing effects compared with RIPC.

Impaired CD27+IgD+ B Cells With Altered Gene Signature in Rheumatoid Arthritis.

Natural antibodies, particularly natural IgM, are proved to play indispensable roles in the immune defenses against common infections. More recently, the protective roles of these natural IgM were also recognized in autoimmune diseases. They are mainly produced by B-1 and innate-like B cells (ILBs). Human CD19+CD27+IgD+ B cells, also termed as un-switched memory B cells, were proposed to be a kind of ILBs. However, functional features and characteristics of these cells in rheumatoid arthritis (RA) remained poorly understood. In this study, we found that human CD27+IgD+ B cells could produce natural antibody-like IgM. Under RA circumstance, the frequencies of these cells were significantly decreased. Moreover, the IgM-producing capacities of these cells were also dampened. Interestingly, the BCR repertoire of these cells was altered in RA, demonstrating decreased diversity with preferential usage alteration from VH3-23D to VH1-8. Single cell sequencing further revealed the proinflammatory biased features of these cells in RA. These CD27+IgD+ B cells were negatively correlated with RA patient disease activities and clinical manifestations. After effective therapy with disease remission in RA, these cells could be recovered. Taken together, these results have revealed that CD27+IgD+ B cells were impaired in RA with dysfunctional features, which might contribute to the disease perpetuation.