Revealing Molecular-Scale Structural Changes in Polymer Nanocomposites during Thermo-Oxidative Degradation Using Evolved Gas Analysis with High-Resolution Time-of-Flight Mass Spectrometry Combined with Principal Component Analysis and Kendrick Mass Defect Analysis.

This study introduces a novel method that utilizes evolved gas analysis with time-of-flight mass spectrometry (EGA-TOFMS) coupled with principal component analysis (PCA) and Kendrick mass defect (KMD) analysis, called EGA-PCA-KMD, to analyze complex structural changes in polymer materials during thermo-oxidative degradation. While EGA-TOFMS captures exact mass data related to the degradation components in the temperature-dependent mass spectra of the evolved products, numerous high-resolution mass spectra with large amounts of ion signals and varying intensities provide challenges for interpretation. To address this, we employed mathematical decomposition through PCA to selectively extract information about the ion series specific to the products that evolved from the degradation components. Additionally, KMD analysis was applied to the attribution of the exact mass signals extracted from the PCA, which categorizes and visualizes depending on the molecular compositions in a two-dimensional plot. The complex structural changes of the triblock copolymer thermoplastic elastomer and its nanocomposites containing nanodiamonds during thermo-oxidative degradation were elucidated using EGA-PCA-KMD to demonstrate the effectiveness of this characterization technique for polymer degradation. Furthermore, it is revealed that the formation of rigid matrix-filler interfacial interaction via the π-π stacking and chemical bonds in the nanocomposites contributes to improvement in the stability toward thermo-oxidative degradation. Our results highlight the benefits of EGA-PCA-KMD and provide valuable insights into polymer degradation.

Poorly Differentiated Chordoma of the Clivus With Loss of SMARCB1 Expression in a Pediatric Patient: A Case Report.

Poorly differentiated chordoma (PDC) is a rare, aggressive subtype of chordoma. A two-year-old girl presented with cervical pain, limb paralysis and respiratory failure. Magnetic resonance imaging and positron emission tomography-computed tomography revealed a tumor compressing the pons at the clivus and osteoblastic metastatic lesions of the left upper arm and right iliac bone. Her tumors shrank substantially after treatment with chemotherapy and proton beam therapy. Our initial diagnosis was an atypical teratoma/rhabdoid tumor, but final diagnosis of PDC was made on the basis of the immunohistochemical expression of brachyury. In addition, the detection of SMARCB1/INI1 mutation confirmed the diagnosis of PDC.

Muscle mass change during chemotherapy in children with high-risk neuroblastoma: a retrospective case series of 24 patients.

The clinical characteristics, cause, and risk factors of sarcopenia are unclear in children. The aim of this study was to describe the course of and identify the factors related to muscle mass change during chemotherapy in children with neuroblastoma. A total of 24 consecutive patients aged below 18 years with newly diagnosed high-risk neuroblastoma between 2010 and 2018 in our hospital were enrolled in a case-series study. The psoas muscle index (PMI) was calculated as a parameter of muscle mass based on computer tomography (CT) images of the psoas muscle. PMIs were evaluated at 4 time points (TPs): TP1, at the diagnosis of neuroblastoma; TP2, after the first cycle of chemotherapy; TP3, after the third cycle of chemotherapy; and TP4, at the end of the induction chemotherapy. PMI recovery was defined as an increase in PMI between TP2 and TP4. The mean PMI decreased by 15% between TP1 and TP2 (TP1 7.09 ± 0.99 vs. TP2 6.01 ± 0.98, P < 0.001) and by 10% between TP1 and TP4 (TP1 7.09 vs. TP4 6.35, P = 0.004). PMI recovery between TP1 and TP2 was observed in 7 (29%) patients. The median age of patients with PMI recovery was significantly lower (2 vs. 4 years, P = 0.028), and the proportion of boys was significantly higher in patients with PMI recovery (100% vs. 41%, P = 0.017).Conclusion: This study demonstrated that prominent PMI reduction occurs during the early time of chemotherapy, and a younger age and male sex may be predictive factors for PMI recovery. What is Known: • Sarcopenia is a common disorder in elderly people. • Several causes and risk factors have been reported in adults. • Children with previous hematological malignancies have decreased physical activity. What is New: • Prominent muscle mass loss was observed early in children with high-risk neuroblastoma during chemotherapy. • Age and sex were found to be potentially associated with muscle mass recovery.

Rapid Fingerprinting of High-Molecular-Weight Polymers by Laser Desorption-Ionization Using Through-Hole Alumina Membrane High-Resolution Mass Spectrometry.

Residual acid found in the desorption ionization using through-holes alumina membranes (DIUTHAME) induces a reproducible protonation/in-source dissociation of polymers made of ester, amide, or siloxane moieties during their surface-assisted laser desorption ionization (SALDI) mass analysis. Deposited on the DIUTHAME chips in solution (solvent-based) or in pure form by melting the polymer powder in situ (solvent-free), high-molecular-weight nylons, silicone, or functionalized celluloses among other polymers are instantly fingerprinted by laser DIUTHAME high-resolution mass spectrometry (MS) with specific patterns resembling their direct analysis in real-time (DART) single-stage or tandem mass spectra. Depending on the polymer, two main types of fingerprints are observed with either the protonated monomer or product ions revealing the nature of the repeating unit or its functionalization. This technique allows a rapid molecular analysis of industrial homopolymers regardless of their molecular weight and complementary to DART with simple or no sample preparation and also promisingly applicable for copolymers.

Molecular characterization of polyethylene oxide based oligomers by surface-assisted laser desorption/ionization mass spectrometry using a through-hole alumina membrane as active substrate.

RATIONALE: Molecular characterization of industrial oligomeric products is performed using surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS), termed desorption ionization using a through-hole alumina membrane (DIUTHAME). This paper describes the unique feature of a DIUTHAME chip applying active SALDI, which generates specific types of fragments of polyglycol samples. METHODS: Polyethylene oxide (PEO) and PEO-based materials were subjected to SALDI-MS. The influence of the presence or absence of a cationization salt on the mass spectrum was investigated. The resulting mass spectra composed of fragment ions were compared with those obtained by collision-induced dissociation (CID)-MS/MS. The specific fragment ions generated using the DIUTHAME chip were further subjected to high-energy CID-MS/MS. RESULTS: The addition of a cationization salt resulted in SALDI mass spectra with fewer fragment peaks. The mass spectra obtained without adding the cationization salt were composed of many more fragment ions caused by in-source decay. The fragmentation pattern was similar to that seen with low-energy CID. The resulting fragment ions were formed by selective cleavage at the C-O bond. High-energy CID-MS/MS can be performed for the specific fragment ions generated by in-source decay fragmentation. CONCLUSIONS: Molecular characterization of PEO-based oligomers by SALDI-MS using the DIUTHAME chip was successfully demonstrated. The selective fragmentation and high-energy CID-MS/MS of the in-source decay fragments made it possible to provide more detailed structural information. This unique feature of DIUTHAME gives it potential for use in new molecular characterization techniques.

A mass spectrometry imaging method for visualizing synthetic polymers by using average molecular weight and dispersity as indices.

RATIONALE: Matrix-assisted laser desorption/ionization mass spectrometric imaging (MSI) is considered to be a powerful tool for visualizing the spatial distribution of synthetic polymers. However, a conventional method extracting an image of a specific m/z value is not suitable for polymers, which have a mass distribution. It is necessary to develop the visualization method to show the spatial distribution of entire polymer series. METHODS: The mass peaks included in polymer series were specified from the average mass spectrum of the entire MSI measurement region by using Kendrick mass defect analysis. The images of those mass peaks were extracted and the number average molecular weight (Mn ), the weight average molecular weight (Mw ) and dispersity (D) were calculated for each pixel. Finally, the spatial distribution of the polymer series was summarized to images using Mn , Mw and D as indices. RESULTS: The effects of the methods were investigated by (i) polymers with different mass distributions and (ii) polymers with different repeat units and end-groups. In both cases, the spatial distribution of specific polymer series including several dozens to hundreds of mass peaks was summarized into three images related to Mn , Mw and D, which are familiar indices in polymer analysis. The results are able to provide an overview of the spatial variation of each polymer more intuitively. CONCLUSIONS: The visualization of Mn , Mw and D will help provide an overview of the spatial distribution of polymer series combined with ion intensity distribution made by conventional methods. It can be also applied to other mass spectrometric imaging methods such as desorption electrospray ionization (DESI) or time-of-flight secondary ion mass spectrometry (TOF-SIMS).

Pharmacokinetic analysis for model-supported therapeutic drug monitoring of busulfan in Japanese pediatric hematopoietic stem cell transplantation recipients.

This prospective observational study analyzed the pharmacokinetics of busulfan in Japanese children and evaluated the predicting accuracy of previous pediatric PPK models of busulfan. This study enrolled five patients (aged 2-12 years, BW 14-48 kg) receiving a busulfan-based conditioning regimen for hematopoietic stem cell transplantation at our hospital between January 2017 and December 2018. All patients received a 2-hour intravenous busulfan infusion four times daily for a total of 16 doses. After the infusions, 51 plasma samples were collected with the plasma busulfan concentration measured by liquid chromatography-tandem mass spectrometry. PPK model fitting was analyzed using the (%MPE) and the (%MAPE). Limited sampling strategies for estimating busulfan AUC were also evaluated. High interpatient variability was observed in the PK parameters. The most suitable PPK model that reflected our data was McCune's two-compartment model (%MPE -8.7, %MAPE 19.3). A combination sampling method using the busulfan concentration at 2 and 6 hours after the start of the first busulfan dose was found to be able to estimate AUC4 day . These results provide useful information on busulfan therapeutic drug monitoring in the Japanese pediatric population.

Graphical Ranking of Divisors to Get the Most out of a Resolution-Enhanced Kendrick Mass Defect Plot.

Resolution-enhanced Kendrick Mass Defect (KMD) analysis using the new concept of fractional base units (repeating unit R divided by integer X; R/ X as a mathematical moiety) is now a powerful data-processing tool to unravel complex mass spectra of polymers. It enhances regular KMD analysis using the chemical moiety, R, to compute mass defects with unprecedented separation of ion series differing by their isotopic or comonomeric contents, end-groups, or charge states in highly visual KMD plots. The value of the divisor, X, dictates the gain of separating power from the regular to the resolution-enhanced KMD plot, and its choice strongly affects the ease and speed of data interpretation. A simple tool to help select the best values of X depending on the users' needs is mandatory to rationalize the analysis and avoid a time-consuming trial-and-error methodology. We propose two graphical representations intuitively ranking the well-suited divisors for the appropriate separation of isotopes or co-oligomers for copolymeric mass-spectral data. Rankings are extended to any type of data set from homopolymeric blends to terpolymers by generalizing the formulas with three variables beyond the specific separation of isotopes. The RANK functions are now available in commercial or homemade spreadsheets (available upon request) to interactively select divisors and compute the associated KMD plots.

microRNA-181a-5p antisense oligonucleotides attenuate osteoarthritis in facet and knee joints.

OBJECTIVES: We recently identified microRNA-181a-5p (miR-181a-5p) as a critical mediator involved in the destruction of lumbar facet joint (FJ) cartilage. In this study, we tested if locked nucleic acid (LNA) miR-181a-5p antisense oligonucleotides (ASO) could be used as a therapeutic to limit articular cartilage degeneration. METHODS: We used a variety of experimental models consisting of both human samples and animal models of FJ and knee osteoarthritis (OA) to test the effects of LNA-miR-181a-5p ASO on articular cartilage degeneration. Histopathological analysis including immunohistochemistry and in situ hybridisation were used to detect key OA catabolic markers and microRNA, respectively. Apoptotic/cell death markers were evaluated by flow cytometry. qPCR and immunoblotting were applied to quantify gene and protein expression. RESULTS: miR-181a-5p expression was increased in human FJ OA and knee OA cartilage as well as injury-induced FJ OA (rat) and trauma-induced knee OA (mouse) cartilage compared with control cartilage, correlating with classical OA catabolic markers in human, rat and mouse cartilage. We demonstrated that LNA-miR-181a-5p ASO in rat and mouse chondrocytes reduced the expression of cartilage catabolic and chondrocyte apoptotic/cell death markers in vitro. Treatment of OA-induced rat FJ or mouse knee joints with intra-articular injections of in vivo grade LNA-miR-181a-5p ASO attenuated cartilage destruction, and the expression of catabolic, hypertrophic, apoptotic/cell death and type II collagen breakdown markers. Finally, treatment of LNA-miR-181a-5p ASO in cultures of human knee OA chondrocytes (in vitro) and cartilage explants (ex vivo) further demonstrated its cartilage protective effects. CONCLUSIONS: Our data demonstrate, for the first time, that LNA-miR-181a-5p ASO exhibit cartilage-protective effects in FJ and knee OA.

Low-dose azacitidine maintenance therapy after allogeneic stem cell transplantation for high-risk pediatric acute myeloid leukemia.

The dismal prognosis of pediatric acute myeloid leukemia (AML) relapsing after hematopoietic stem cell transplantation (HSCT) requires exploration of novel strategies to prevent relapse. Azacitidine (AZA) maintenance therapy could potentially reduce the recurrence rate post HSCT. Here, we presents the cases of three children with high-risk AML post HSCT who were treated with low-dose AZA maintenance therapy, demonstrating the feasibility of this therapy. Currently, all three are in complete remission for 13-41 months despite their high-risk characteristics. Our encouraging data warrant larger prospective studies to assess the efficacy and safety of low-dose AZA maintenance therapy post HSCT for pediatric patients with high-risk AML.

Predicting Heparin Responsiveness in Children Before Cardiopulmonary Bypass: A Retrospective Cohort Study.

BACKGROUND: Inadequate or excess administration of unfractionated heparin for cardiopulmonary bypass (CPB) can cause significant harm. Age-dependent differences in the pharmacodynamics and pharmacokinetics of heparin contribute to increased variability of heparin responsiveness in children. The aims of the current study were to (1) examine the correlation between predicted and observed heparin responsiveness in children before CPB measured using the Hemostasis Management System (HMS) Plus (Medtronic, Minneapolis, MN), (2) describe age-specific reference intervals for heparin sensitivity index (HSI) observed in children, and (3) test predictive models of HSI using preoperative clinical and laboratory data. METHODS: In this retrospective cohort study, children (ages ≤17 years) who required therapeutic heparinization for CPB in a 40-month period between September 2010 and December 2013 were investigated. Children weighing ≥45 kg or with a height ≥142 cm were excluded. HSI was defined as the difference between activated clotting time after heparin administration and the baseline activated clotting time divided by the heparin-loading dose (IU) per kilogram. Lin's concordance correlation coefficient was used for the primary analysis of the relationship between predicted and observed HSI. Reference intervals were calculated for HSI using medians and 2.5% and 97.5% percentiles according to established guidelines for clinical and laboratory standards. Nonparametric regression analyses were used to model the relationship between HSI (dependent variable) and preoperative covariates (independent variables). RESULTS: A total of 1281 eligible children were included in the final analysis. Overall, there was a moderate correlation between predicted and observed HSI measured using HMS Plus System (rho_c = 0.46; 95% confidence interval, 0.41-0.50; P < .001). Sixty-five percent (829 of 1281) of predicted HSI values were less than observed. From adjusted regression models, HSI was best predicted by preoperative international normalized ratio, platelet count, and weight, but this model accounted for only 25% of the variance in HSI. CONCLUSIONS: In a large cohort of children, heparin responsiveness before CPB was not reliably predicted by either in vitro measurement using the HMS Plus System or commonly available preoperative clinical and laboratory data. We describe age-specific reference intervals for HSI in children, and we anticipate that these data will aid the identification of heparin resistance in this population.

Ribosomal subunit protein typing using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) for the identification and discrimination of Aspergillus species.

BACKGROUND: Accurate identification of Aspergillus species is a very important subject. Mass spectral fingerprinting using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) is generally employed for the rapid identification of fungal isolates. However, the results are based on simple mass spectral pattern-matching, with no peak assignment and no taxonomic input. We propose here a ribosomal subunit protein (RSP) typing technique using MALDI-TOF MS for the identification and discrimination of Aspergillus species. The results are concluded to be phylogenetic in that they reflect the molecular evolution of housekeeping RSPs. RESULTS: The amino acid sequences of RSPs of genome-sequenced strains of Aspergillus species were first verified and compared to compile a reliable biomarker list for the identification of Aspergillus species. In this process, we revealed that many amino acid sequences of RSPs (about 10-60%, depending on strain) registered in the public protein databases needed to be corrected or newly added. The verified RSPs were allocated to RSP types based on their mass. Peak assignments of RSPs of each sample strain as observed by MALDI-TOF MS were then performed to set RSP type profiles, which were then further processed by means of cluster analysis. The resulting dendrogram based on RSP types showed a relatively good concordance with the tree based on ß-tubulin gene sequences. RSP typing was able to further discriminate the strains belonging to Aspergillus section Fumigati. CONCLUSIONS: The RSP typing method could be applied to identify Aspergillus species, even for species within section Fumigati. The discrimination power of RSP typing appears to be comparable to conventional ß-tubulin gene analysis. This method would therefore be suitable for species identification and discrimination at the strain to species level. Because RSP typing can characterize the strains within section Fumigati, this method has potential as a powerful and reliable tool in the field of clinical microbiology.

MALDI SpiralTOF high-resolution mass spectrometry and Kendrick mass defect analysis applied to the characterization of poly(ethylene-co-vinyl acetate) copolymers.

RATIONALE: Poly(ethylene-co-vinyl acetate) copolymers - usually referred to as EVA - are first class industrial polymers used for applications ranging from padding to photovoltaics as encapsulant for the silicon solar cells. Various techniques have been used for their characterization but the analysis of intact EVA chains using mass spectrometry (MS) has not been reported so far. METHODS: Three copolymers containing 18, 25 and 40 wt% vinyl acetate (VA) have been characterized using an off-line coupling of size-exclusion chromatography (SEC) and matrix-assisted laser desorption/ionization (MALDI) spiral-time-of-flight (TOF) high-resolution mass spectrometry (HRMS). The representativeness of those results for the entire samples has been checked using (13) C NMR spectroscopy. Lastly, Kendrick mass defect analysis has been proposed as an alternative and user-friendly data treatment method. RESULTS: The shortest chains isolated by SEC fractionation and mass-analyzed by HRMS have been thoroughly described in terms of end-groups (found to be hydrogens) and co-monomeric composition. The VA content was successfully derived from the peak assignments in MS spectra for the EVA 40 wt% and 25 wt% while it tended to be overestimated for the latest EVA 18 wt% (increasing poly(ethylene) character). Similar results have been found using a faster data treatment method relying on the Kendrick mass defect analysis of the MS data. CONCLUSIONS: EVA low molecular weight intact oligomers have been extensively characterized by MS for the first time and the structural features confidently extended to the full sample according to NMR data. The Kendrick mass analysis finally constituted an efficient method for a fast evaluation of their VA content with no need for manual assignment. © 2016 The Authors. Rapid Communications in Mass Spectrometry Published by John Wiley & Sons Ltd.

[Evaluation of the pharmaceutical stability of polaprezinc/sodium alginate gargle solution containing lidocaine].

OBJECTIVE: A gargle solution(L-P/AG)for the treatment of painful stomatitis was prepared by adding lidocaine to a polaprezinc/sodium alginate gargle solution(P/AG), and its pharmaceutical stability was evaluated. METHODS: L-P/AG was stored at 5, 25, and 40°C. The strengths of polaprezinc and lidocaine were determined. The viscosity and pH of L-P/AG were also determined, and its appearance was evaluated. RESULTS: When stored at 5 or 25°C in a dark place, L-P/AG showed neither reduction in the strength of either drug nor did it show a change in the viscosity, pH, or appearance. When stored exposed to light at 40°C, L-P/AG showed reductions in the strength of both drugs, as well as in viscosity and pH; furthermore, a change in appearance was noted. DISCUSSION: L-P/AG prepared for the treatment of painful stomatitis remains pharmaceutically stable for 28 days when stored at 25°C in a dark place.

Injectable bone tissue engineering using expanded mesenchymal stem cells.

Patients suffering from bone defects are often treated with autologous bone transplants, but this therapy can cause many complications. New approaches are therefore needed to improve treatment for bone defects, and stem cell therapy presents an exciting alternative approach. Although extensive evidence from basic studies using stem cells has been reported, few clinical applications using stem cells for bone tissue engineering have been developed. We investigated whether injectable tissue-engineered bone (TEB) composed of mesenchymal stem cells (MSCs) and platelet-rich plasma was able to regenerate functional bone in alveolar deficiencies. We performed these studies in animals and subsequently carried out large-scale clinical studies in patients with long-term follow-up; these showed good bone formation using minimally invasive MSC transplantation. All patients exhibited significantly improved bone volume with no side effects. Newly formed bone areas at 3 months were significantly increased over the preoperation baseline (p < .001) and reached levels equivalent to that of native bone. No significant bone resorption occurred during long-term follow-up. Injectable TEB restored masticatory function in patients. This novel clinical approach represents an effective therapeutic utilization of bone tissue engineering.

Income Level and Impaired Kidney Function Among Working Adults in Japan.

Importance: Chronic kidney disease (CKD) is a major public health issue, affecting 850 million people worldwide. Although previous studies have shown the association between socioeconomic status and CKD, little is known about whether this association exists in countries such as Japan where universal health coverage has been mostly achieved. Objective: To identify any association of income-based disparity with development of impaired kidney function among the working population of Japan. Design, Setting, and Participants: This was a nationwide retrospective cohort study of adults aged 34 to 74 years who were enrolled in the Japan Health Insurance Association insurance program, which covers approximately 40% of the working-age population (30 million enrollees) in Japan. Participants whose estimated glomerular filtration rate (eGFR) had been measured at least twice from 2015 to 2022 were included in the analysis, which was conducted from September 1, 2021, to March 31, 2023. Exposure: Individual income levels (deciles) in the fiscal year 2015. Main Outcomes and Measures: Odds ratios were calculated for rapid CKD progression (defined as an annual eGFR decline of more than 5 mL/min/1.73 m2), and hazard ratios, for the initiation of kidney replacement therapy (dialysis or kidney transplant) by income level deciles in the fiscal year 2015. Results: The study population totaled 5 591 060 individuals (mean [SD] age, 49.2 [9.3] years) of whom 33.4% were female. After adjusting for potential confounders, the lowest income decile (lowest 10th percentile) demonstrated a greater risk of rapid CKD progression (adjusted odds ratio, 1.70; 95% CI, 1.67-1.73) and a greater risk of kidney replacement therapy initiation (adjusted hazard ratio, 1.65; 95% CI, 1.47-1.86) compared with the highest income decile (top 10th percentile). A negative monotonic association was more pronounced among males and individuals without diabetes and was observed in individuals with early (CKD stage 1-2) and advanced (CKD stage 3-5) disease. Conclusions and Relevance: The findings of this retrospective cohort study suggest that, even in countries with universal health coverage, there may be a large income-based disparity in the risk of rapid CKD progression and initiation of kidney replacement therapy. These findings highlight the importance of adapting CKD prevention and management strategies according to an individual's socioeconomic status, even when basic health care services are financially guaranteed.

HIF-1α and MIF enhance neutrophil-driven type 3 immunity and chondrogenesis in a murine spondyloarthritis model.

The hallmarks of spondyloarthritis (SpA) are type 3 immunity-driven inflammation and new bone formation (NBF). Macrophage migration inhibitory factor (MIF) was found to be a key driver of the pathogenesis of SpA by amplifying type 3 immunity, yet MIF-interacting molecules and networks remain elusive. Herein, we identified hypoxia-inducible factor-1 alpha (HIF1A) as an interacting partner molecule of MIF that drives SpA pathologies, including inflammation and NBF. HIF1A expression was increased in the joint tissues and synovial fluid of SpA patients and curdlan-injected SKG (curdlan-SKG) mice compared to the respective controls. Under hypoxic conditions in which HIF1A was stabilized, human and mouse neutrophils exhibited substantially increased expression of MIF and IL-23, an upstream type 3 immunity-related cytokine. Similar to MIF, systemic overexpression of IL-23 induced SpA pathology in SKG mice, while the injection of a HIF1A-selective inhibitor (PX-478) into curdlan-SKG mice prevented or attenuated SpA pathology, as indicated by a marked reduction in the expression of MIF and IL-23. Furthermore, genetic deletion of MIF or HIF1A inhibition with PX-478 in IL-23-overexpressing SKG mice did not induce evident arthritis or NBF, despite the presence of psoriasis-like dermatitis and blepharitis. We also found that MIF- and IL-23-expressing neutrophils infiltrated areas of the NBF in curdlan-SKG mice. These neutrophils potentially increased chondrogenesis and cell proliferation via the upregulation of STAT3 in periosteal cells and ligamental cells during endochondral ossification. Together, these results provide supporting evidence for an MIF/HIF1A regulatory network, and inhibition of HIF1A may be a novel therapeutic approach for SpA by suppressing type 3 immunity-mediated inflammation and NBF.