Itaconate and dimethyl itaconate upregulate IL-6 production in the LPS-induced inflammation in mice.

Itaconate is one of the most studied immunometabolites produced by myeloid cells during inflammatory response. It mediates a wide range of anti-inflammatory and immunoregulatory effects and plays a role in a number of pathological states, including autoimmunity and cancer. Itaconate and its derivatives are considered as potential therapeutic agents for treatment of inflammatory diseases. While immunoregulatory effects of itaconate have been extensively studied in vitro and using knock-out mouse models, less is known about how therapeutic administration of this metabolite regulates inflammatory response in vivo. Here, we investigate the immunoregulatory properties of exogenous administration of itaconate (ITA) and its derivative dimethyl itaconate (DI) in a mouse model of LPS-induced inflammation. The data show that administration of ITA or DI controls systemic production of multiple cytokines, including increased IL-10 production. However, only DI was able to suppress systemic production of IFNγ and IL-1ß. In contrast to in vitro data, administration of ITA or DI in vivo resulted in systemic upregulation of IL-6 in the blood. Electrophilic stress due to ITA or DI was not responsible for IL-6 upregulation. However, inhibition of SDH with dimethyl malonate (DM) also resulted in elevated systemic levels of IL-6 and IL-10. Taken together, our study reports a novel effect of exogenous itaconate and its derivative DI on the production of IL-6 in vivo, with important implications for the development of itaconate-based anti-inflammatory therapies.

The immunometabolite itaconate stimulates OXGR1 to promote mucociliary clearance during the pulmonary innate immune response.

Pathogens and inflammatory conditions rapidly induce the expression of immune-responsive gene 1 (IRG1) in cells of myeloid lineage. IRG1 encodes an aconitate decarboxylase (ACOD1) that produces the immunomodulatory metabolite itaconate (ITA). In addition to rapid intracellular accumulation, ITA is also secreted from the cell, but whether secreted ITA functions as a signaling molecule is unclear. Here, we identified ITA as an orthosteric agonist of the GPCR OXGR1, with an EC50 of approximately 0.3 mM, which was in the same range as the physiological concentration of extracellular ITA upon macrophage activation. ITA activated OXGR1 to induce Ca2+ mobilization, ERK phosphorylation, and endocytosis of the receptor. In a mouse model of pulmonary infection with bacterial Pseudomonas aeruginosa, ITA stimulated Oxgr1-dependent mucus secretion and transport in respiratory epithelium, the primary innate defense mechanism of the airway. Our study thus identifies ITA as a bona fide ligand for OXGR1 and the ITA/OXGR1 paracrine signaling pathway during the pulmonary innate immune response.

Diagnostic Accuracy of the Synovial Fluid α-Defensin Lateral Flow Test in Periprosthetic Joint Infection: A Meta-analysis.

OBJECTIVES: There is a controversy on the diagnostic reliability and accuracy of synovial fluid α-defensin in periprosthetic joint infection (PJI). We performed this meta-analysis to evaluate the diagnostic accuracy of the α-defensin lateral flow test in PJI. METHODS: PubMed, Embase, and the Cochrane library were systematically searched, and articles (up to January 2020) on the diagnosis of hip and knee PJIs using the α-defensin Synovasure lateral flow test were included. The diagnostic accuracy of the α-defensin lateral flow test in PJI was evaluated using meta-analysis. The pooled sensitivity, specificity, accuracy, positive and negative likelihood ratio, diagnostic odds ratio, and post-test probabilities were calculated. RESULTS: Seventeen studies including 1443 cases were included. Meta-analysis showed the pooled sensitivity, specificity, positive likelihood ratio, negative likelihood ratio, and a diagnostic odds ratio was 0.83 (95% CI 0.77, 0.88), 0.95 (95% CI 0.93, 0.97), 16.86 (95% CI 11.67, 24.37), 0.17 (95% CI 0.13, 0.24) and 85.30 (95% CI 47.76, 152.35), respectively. The area under the hierarchical summary receiver operating characteristic curve was 0.97 (95% CI 0.95, 0.98). Subgroup analysis also confirmed the high efficiency of α-defensin Synovasure lateral flow test in diagnosing PJIs, irrespective of ethnicity. Fagan's nomogram analysis there was a high positive post-test probability of 94% and a low negative post-test probability of 15%. CONCLUSIONS: We indicated that the α-defensin lateral flow test had a high accuracy for diagnosing PJI. Large-scale studies are needed to validate its significance in PJI diagnosis.

Tumor-derived neomorphic mutations in ASXL1 impairs the BAP1-ASXL1-FOXK1/K2 transcription network.

Additional sex combs-like 1 (ASXL1) interacts with BRCA1-associated protein 1 (BAP1) deubiquitinase to oppose the polycomb repressive complex 1 (PRC1)-mediated histone H2A ubiquitylation. Germline BAP1 mutations are found in a spectrum of human malignancies, while ASXL1 mutations recurrently occur in myeloid neoplasm and are associated with poor prognosis. Nearly all ASXL1 mutations are heterozygous frameshift or nonsense mutations in the middle or to a less extent the C-terminal region, resulting in the production of C-terminally truncated mutant ASXL1 proteins. How ASXL1 regulates specific target genes and how the C-terminal truncation of ASXL1 promotes leukemogenesis are unclear. Here, we report that ASXL1 interacts with forkhead transcription factors FOXK1 and FOXK2 to regulate a subset of FOXK1/K2 target genes. We show that the C-terminally truncated mutant ASXL1 proteins are expressed at much higher levels than the wild-type protein in ASXL1 heterozygous leukemia cells, and lose the ability to interact with FOXK1/K2. Specific deletion of the mutant allele eliminates the expression of C-terminally truncated ASXL1 and increases the association of wild-type ASXL1 with BAP1, thereby restoring the expression of BAP1-ASXL1-FOXK1/K2 target genes, particularly those involved in glucose metabolism, oxygen sensing, and JAK-STAT3 signaling pathways. In addition to FOXK1/K2, we also identify other DNA-binding transcription regulators including transcription factors (TFs) which interact with wild-type ASXL1, but not C-terminally truncated mutant. Our results suggest that ASXL1 mutations result in neomorphic alleles that contribute to leukemogenesis at least in part through dominantly inhibiting the wild-type ASXL1 from interacting with BAP1 and thereby impairing the function of ASXL1-BAP1-TF in regulating target genes and leukemia cell growth.

[Advances on treatment of periprosthetic infection and antibiotic delivery system after total hip arthroplasty].

Periprosthetic infection after hip replacement is a clinical catastrophic disease, which often leads to the failure of the prosthesis. It needs the combination of systemic antibiotics to cure the infection, which brings huge burden to doctors and patients. There are strict indications for debridement and one-stage revision of the prosthesis, and few cases meet the requirements. The second revision is still the gold standard for the treatment of periprosthetic infection. It is suitable for all infection conditions and has a high success rate. On the second phase of renovation, the antibiotic sustained release system plays a key role, and the carrier of antibiotic sustained-release system is the focus of current research, including classic bone cement and absorbable biomaterials. Bone cement has strong mechanical strength, but the antibiotic release shows a sharp decline trend; the absorbable biomaterials can continuously release antibiotics with high concentration, but the mechanical strength is poor, so it could not use alone. The combination of bone cement and absorbable biomaterials will be an ideal antibiotic carrier. PMMA is the most commonly used antibiotic carrier, but the antibiotic release concentration is decreased sharply after 24 hours. It will be difficult to control the infection and increase the risk of bacterial resistance if it is lower than the minimum inhibitory concentration. The biodegradable materials can release antibiotics completely, with long release time and high concentration, but low mechanical strength. Antibiotic spacer plays an important role in the control of infection. In the future, how to further extend the antibiotic release time of antibiotic sustained-release system, increase the amount of antibiotic release and maintain the mechanical strength of the material will be studied.

[Procalcitonin as a sensitive index of chronic infection after artificial joint replacement:a retrospective analysis].

OBJECTIVE: To study the predictive value of procalcitonin as a serum biomarker in diagnosis of late periprosthetic joint infection(PJI) for providing theoretical reference basis for diagnosis of PJI. METHODS: A total of 77 cases were retrospective included from January 2015 to December 2017 for revision of total hip arthroplasty and total knee arthroplasty, according to the diagnostic criteria of Musculoskeletal Infection Society(MISI). All cases were divided into infection group and non-infection group. Infection group included 21 cases, 7 cases for male and 14 cases for female, with an average age of (60.70±8.75) years old (ranged 43 to 75 years old). Non-infection group 56 included cases, 24 cases for male and 32 cases for female, with an average age (64.40±12.14) years old (ranged 43 to 85 years old). Concentration of preoperative serum procalcitonin was examined and the chi-square test was used to compare positive rate between the two groups. RESULTS: Two cases in the infection group had positive serum procalcitonin, 0.06 ng/L and 0.10 ng/L respectively, with the positive rate of 9.52%; 4 cases in non-infected group had positive serum procalcitonin, 0.05 ng/L, 0.06 ng/L, 0.06 ng/L, 0.16 ng/L respectively, with the positive rate of 7.14%. No statistically significant difference was observed between the two groups (P=0.662). CONCLUSIONS: Most of PJI were low toxicity infection with little systemic inflammatory response, so the concentration of serum procalcitonin was normal or slightly higher level, had little clinical significance for diagnosis of PJI. But the cases of this retrospective study are not enough, more cases are needed for further study.

Does serum calcium relate to different types of hip fracture? A retrospective study.

PURPOSE: To investigate the potential correlation between two different types of hip fractures and serum calcium levels. METHODS: We consecutively studied 101 cases of femoral neck fracture and 95 cases of femoral inter- trochanteric fracture between January 2011 and December 2013. Fasting blood samples were taken and serum calcium measurements were performed respectively in three periods: the time of admission, postoperation, and discharge. Creatinine, alkaline phosphatase and albumin were also analyzed. RESULTS: Considering the levels of serum calcium between two groups at the time of admission, post- operation and discharge, there was significant difference at admission and discharge (p <0.05), while there was no significant difference at the time of postoperation (p > 0.05). The magnitude of serum calcium fluctuation was larger in femoral neck group than femoral intertrochanteric group. Concerning alkaline phosphatase and albumin levels at admission, there was no significant difference between two groups (p > 0.05). CONCLUSION: The capability of reservation and restoration of serum calcium in patients with femoral neck fracture is better than that in patients with femoral intertrochanteric fracture. A low serum calcium level may be susceptible to femoral intertrochanteric fracture.

Vascularised greater trochanter bone graft, combined free iliac flap and impaction bone grafting for osteonecrosis of the femoral head.

PURPOSE: To investigate the curative efficacy of osteonecrosis of the femoral head (ONFH) in a hip-preserving operative approach, by grafting a vascularized greater trochanter flap combined with a free iliac flap, in an attempt to seek an innovative approach for patients who suffered middle to late stage ONFH without total hip arthroplasty (THA) surgery. METHOD: Our research included a total of 60 patients (66 hips) who accepted hip-preserving surgery by grafting a vascularized greater trochanter flap combined with a free iliac flap which was tightly filled by hammering because of ONFH (most were Association Research Circulation Osseous (ARCO) stage III patients) from January, 2006 to December, 2010. A Harris Hip Score was obtained during follow-ups, evaluating the clinical efficacy, X-rays were taken regularly for image assessing, and the SF-36 scale was used for estimating quality of life. Terminal observation time was considered when patients had symptom-dependant indications for performing another hip-preserving surgery or THA surgery. RESULTS: Fifty-eight patients (64 hips) were eventually contacted by telephone for an out-patient clinic return visit, with a mean follow-up time of 35.8 months (varied from 12 months to 60 months), but two patients lost contact for various reasons. The demographic data were as follows: there were 16 ARCO IIIA cases, 22 ARCO IIIB cases, and 26 ARCO IIIC cases, respectively. Postoperative X-rays revealed a well-repaired necrotic area of the femoral head and improvement of femoral-acetabulum coverage. The last follow-up mean Harris Hip Score was 86.56 ± 7.38 (excellent results reached 87.50%), which were greatly improved compared to 50.95 ± 6.86 pre-operatively. Also the postoperative mean scores of all dimensions of the SF-36 scale were improved to some extent. Additionally the physical component summary (PSC) scores were enhanced from 42 ± 13 pre-operatively to 78 ± 11, while the postoperative mental component summary (MCS) scores (76 ± 11) largely increased in contrast to pre-operative scores (51 ± 10), with both target indices having statistical significance (p = 0.005, p = 0.01), signifying hugely improvement of the quality of life of the patients. A correlation was found between Harris Hip Score and all dimensions of SF-36 scale (r = 0.32-0.72), especially closely correlated with physical functioning (PF), role-physical (RP) and bodily pain (BP) in PCS aspect (r = 0.72, p < 0.01; r = 0.58, p < 0.01; r = 0.65, p < 0.01, respectively). CONCLUSION: There is definite curative efficacy for the treatment of ONFH with an hip-preserving operative approach by grafting a vascularized greater trochanter flap combined with a free iliac flap which was tightly filled by hammering. This kind of operative approach reconstructs the biological stability of femoral head, which promotes repair of necrotic areas and indirectly preserves the femoral head of patients and a majority of hip function. It possesses vast clinical as well as practical significance, because the long-term efficacy can satisfy fundamental life requirements, especially for those young and middle-aged patients who suffer ONFH to avoid or put off the time of total hip arthroplasty (THA) surgery.

[Experimental study of directional differentiation of bone mesenchymal stem cells (BMSCs) to osteoblasts guided by serum containing cistanche deserticola].

OBJECTIVE: To study directional differentiation of BMSCs guided by Desert living Cistanche (Herba Cistanches) which invigorates the kidney. METHODS: Primary BMSCs were obtained by whole bone marrow culture and subcultured to the fourth generation by trypsin digestion, and than inoculated into two six-well plates at 5 x 10(6) cells per milliliter, all the plates were divided into three groups as blank group, Dexamethasone (DXM) group and Herba Cistanches group, three wells in each group, medium were changed at day 2. The blank group were changed with L-DMEM containing 10% FBS. The DXM group were changed with medium containing 10 mmol/L beta-sodium glycerophosphate, 0.1 micromol/L DXM and 50 mg/L vitamin C. The Herba Cistanches group were changed with medium containing 10% blood serum containing Herba Cistanches and L-DMEM. One of the six-well plates was stained by alkaline phosphatase (AKP) at the tenth day,the other one was stained by alizarin Bordeaux at the twentieth day. RESULTS: At the tenth day DXM group and Herba Cistanches group were ALKP stained positive; from the 12th day,white calcium nodus could be seen at the surface of the wells; which alizarin stained positive by the twentyth day. CONCLUSION: The medium containing Herba Cistanches can guide BMSCs to differentiate into osteoblast, which promises a favorable prospect for the treatment of osteoporosis and bone fracture disunion.

[Human osteoblast-like cells OS-732 intervened with alcohol and treated with liuweidihuang pill medicated serum].

OBJECTIVE: To investigate the mechanism of osteonecrosis of the femoral head caused by alcohol and the therapeutic effects of the method of invigorating the kidney. METHODS: The osteoblast-like cells were intervened with 0.3, 0.9, 1.5 mol/L alcohol and observed by flow cytometry and light microscopy and electro microscopy. The cells intervened by 0.15 mol/L alcohol were treated with liuweidihuang pill medicated serum and observed by flow cytometry and light microscopy and electro microscopy. RESULTS: Apoptosis rate and the degree of cell necrosis had positive correlation with the concentration and the acting time of alcohol. The polymorphism of cells wer turned into roundness, and the cellular ecphyma was shortened and thinning, ribosome and endocytoplasmic reticulum were diminished. Part of the ruffle in mitochondria was disappeared. After treatment, the cellular ecphyma was recovered and the endoplasmic reticulum and golgi apparatus were visible. CONCLUSION: The osteoblast-like cells could be necrosis and apoptosis by the alcohol and were positive correlation with the intervened concentration and acting time. Chinese medicine with method of invigorating the kidney has effect on rehabilitating the cellular activity.