Immune cell profiling in the joint following human and murine articular fracture.

OBJECTIVE: Human and in vivo animal research implicates inflammation following articular fracture as contributing to post-traumatic arthritis. However, relevant immune cell subsets present following injury are currently undefined. Immunophenotyping human and murine synovial fluid may help to identify immune cell populations that play key roles in the response to articular fracture. METHODS: Immunophenotyping by polychromatic flow cytometry was performed on human and mouse synovial fluid following articular fracture. Specimens were collected in patients with closed ankle fracture at the time of surgical fixation and from C57BL/6 mice with closed articular knee fracture. Immune cells were collected from injured and uninjured joints in mice via a novel cell isolation method. Whole blood samples were also collected. Immunohistochemistry (IHC) was performed on mouse synovial tissue to assess for macrophages and T cells. RESULTS: Following intra-articular fracture, the prominent human synovial fluid immune cell subset was CD3+ T cells, containing both CD4+ and CD8+ T cells. In mice, infiltration of CD45+ immune cells in synovial fluid of the fractured limb was dominated by CD19+ B cells and CD3+ T cells at 7 days after intra-articular fracture. We also detected adaptive immune cells, including macrophages, NK cells, dendritic cells and monocytes. Macrophage and T cell findings were supported by IHC of murine synovial tissue. CONCLUSIONS: Determining specific cell populations that mediate the immune response is essential to elucidating the chain of events initiated after injury and may be an important step in identifying potential immune signatures predictive of PTA susceptibility or potential therapeutic targets.

An Asian perspective on improving outcomes for nasal bone fractures by establishing specific treatment options.

OBJECTIVE: To report treatment outcomes of patients with different types of nasal bone fracture, following a tailored treatment protocol. DESIGN: The patterns and the severity of the fractures were determined by a preoperative facial photo and nasal bone computed tomography (CT) retrospectively. SETTING: A tertiary referral centre. PARTICIPANTS: We evaluated 129 patients who underwent surgery between March 2002 and January 2014. Patients were subjected to five different treatment methods depending on the severity of injury. MAIN OUTCOME MEASURES: Medical records were reviewed to assess rates of complications and revision surgery. Cosmetic and functional (the degree of nasal obstruction) outcomes were measured using the Likert scale from 1 (very dissatisfied, severe obstruction) to 5 (very satisfied, no obstruction). RESULTS: The mean elapsed time from injury to surgery was 14.9 days. The overall treatment failure rate, defined as a deformity that required revision, was 6.2%. The patient satisfaction scores for aesthetic and functional outcomes were 3.9 and 4.6 respectively. CONCLUSION: The prudent selection of patients with indication for closed reduction, and further specification of variable treatment options for the various degrees of nasal bone fracture, may be helpful in achieving an improved treatment outcome. LEVEL OF EVIDENCE: level IV.

Characterization of CD4 and CD8 T cell responses in MuSK myasthenia gravis.

Muscle specific tyrosine kinase myasthenia gravis (MuSK MG) is a form of autoimmune MG that predominantly affects women and has unique clinical features, including prominent bulbar weakness, muscle atrophy, and excellent response to therapeutic plasma exchange. Patients with MuSK MG have predominantly IgG4 autoantibodies directed against MuSK on the postsynaptic muscle membrane. Lymphocyte functionality has not been reported in this condition. The goal of this study was to characterize T cell responses in patients with MuSK MG. Intracellular production of IFN-gamma, TNF-alpha, IL-2, IL-17, and IL-21 by CD4+ and CD8+ T cells was measured by polychromatic flow cytometry in peripheral blood samples from 11 Musk MG patients and 10 healthy controls. Only one MuSK MG patient was not receiving immunosuppressive therapy. Regulatory T cells (Treg) were also included in our analysis to determine if changes in T cell function were due to altered Treg frequencies. CD8+ T cells from MuSK MG patients had higher frequencies of polyfunctional responses than controls, and CD4+ T cells had higher IL-2, TNF-alpha, and IL-17. MuSK MG patients had a higher percentage of CD4+ T cells producing combinations of IFN-gamma/IL-2/TNF-gamma, TNF-alpha/IL-2, and IFN-gamma/TNF-alpha. Interestingly, Treg numbers and CD39 expression were not different from control values. MuSK MG patients had increased frequencies of Th1 and Th17 cytokines and were primed for polyfunctional proinflammatory responses that cannot be explained by a defect in CD39 expression or Treg number.

A model for pregnancy rates after IVF-ET in patients with infertility and endometriosis.

OBJECTIVE: The objective of the present study was to examine the clinical factors influencing the pregnancy rate of infertile patients with endometriosis and establish a predictive model. PATIENTS AND METHODS: This study included 158 patients (158 cycles) with infertility and endometriosis who underwent laparoscopic surgery, and in vitro fertilization and embryo transfer (IVF-ET) were evaluated retrospectively between January 2019 and December 2020. The clinical factors in the pregnant and non-pregnant group were analyzed by univariate analysis. Statistically significant variables were subsequently used for multivariate logistic regression to establish the prediction model. RESULTS: Multivariate logistic regression analyses showed that GnRH-a treatment after operation (OR, 6.562; 95% CI: 2.782-15.477; p<0.01), ASRM stage (OR, 0.218; 95% CI: 0.093-0.509; p<0.05), the number of high-quality transferred embryos (OR, 3.155; 95% CI: 1.647-6.047; p<0.05) were independently associated with successful pregnancy. The area under the curve (AUC) of the prediction model was 0.774 (95% CI: 0.700-0.847). According to Hosmer-Lemeshow, the model was well fitted (p>0.05). We applied the bootstrapping method to internal validation, and the result showed that the pregnancy rate predicted by the model and the real data were consistent. CONCLUSIONS: The models for predicting pregnancy rates after IVF-ET in infertility and endometriosis patients showed high accuracy. The effective methods to increase the number of high-quality embryos need further study.

The activation of B-Raf and Raf-1 after electroconvulsive shock in the rat hippocampus.

We demonstrated that ECS activates the kinase activity of B-Raf and Raf-1 in the rat hippocampus. The activity was maximal at one minute after ECS and temporally coincided with the increased membrane translocation of Rafs and the reported activity of MAPK, but not with the phosphorylation of Rafs.

TRIM72 negatively regulates myogenesis via targeting insulin receptor substrate-1.

Lipid rafts have been known to be platforms to initiate cellular signal transduction of insulin-like growth factor (IGF) inducing skeletal muscle differentiation and hypertrophy. Here, tripartite motif 72 (TRIM72), with a really interesting new gene (RING)-finger domain, a B-box, two coiled-coil domains, and a SPRY (SPla and RYanodine receptor) domain, was revealed to be predominantly expressed in the sarcolemma lipid rafts of skeletal and cardiac muscles. Adenoviral TRIM72 overexpression prevented but RNAi-mediated TRIM72 silencing enhanced C2C12 myogenesis by modulating the IGF-induced insulin receptor substrate-1 (IRS-1) activation through the molecular association of TRIM72 with IRS-1. Furthermore, myogenic activity was highly enhanced with increased IGF-induced Akt activation in the satellite cells of TRIM72(-/-) mice, compared to those of TRIM72+/+ mice. Because TRIM72 promoter analysis shows that two proximal E-boxes in TRIM72 promoter were essential for MyoD- and Akt-dependent TRIM72 transcription, we can conclude that TRIM72 is a novel antagonist of IRS-1, and is essential as a negative regulator of IGF-induced muscle differentiation.

The influence of carotid artery catheterization technique on the incidence of thromboembolism during carotid artery stenting.

BACKGROUND AND PURPOSE: Thromboembolic events related to CAS continue to be the main limitation to the widespread use of this technique as a first-line treatment for carotid occlusive disease. Our aim was to evaluate thromboembolism during CAS using DWI for catheterization techniques of the carotid artery. MATERIALS AND METHODS: Thirty-two consecutive patients with symptomatic carotid stenosis underwent CAS involving 1 of 2 carotid artery catheterization techniques: One used a 7F or 8F catheter (group 1, n = 16) and the other used a coaxial system in which a 7F or 8F catheter was used in conjunction with a 4F or 5F catheter (group 2, n = 16). DWI was performed before and after CAS. Clinical variables, the number and location of NES on DWI after CAS, were compared between the 2 groups. RESULTS: NES on DWI occurred in 53% of all patients. The incidence of NES was significantly higher in patients 65 years of age and older versus those younger than 65 years of age (P = .013). All NESs were asymptomatic, and their rate of occurrence did not differ significantly between groups 1 and 2. The incidence of NES in the other territories that were outside that of the treated carotid artery (P = .004) and the incidence of multiple NESs (P = .04) were significantly higher in group 1. CONCLUSIONS: NES in the other territories mainly arises from the atherosclerotic aortic arch and arch vessels during the manipulation of endoluminal devices. The carotid artery catheterization technique using the coaxial system with a 7F or 8F catheter in conjunction with a 4F or 5F catheter reduced the incidence of NES in the other territories.

Analysis of the results of the micronucleus test in patients presenting upper digestive tract cancers and in non-cancerous subjects.

A micronucleus test was performed on 75 subjects of whom 38 presented with cancer of the upper digestive tract and 37 were free of disease; the absence of cancerous or pre-cancerous lesions in this latter group was confirmed by endoscopy and vital staining. The daily levels of alcohol and tobacco consumption of the 75 subjects were determined by precise questioning: 78% of the non-cancerous subjects smoked less than 10 g of tobacco per day whereas 79% of the cancer patients smoked 10 g or more daily. The alcohol intake of 78% of the non-cancerous subjects and 63% of the cancer patients was less than 101 ml per day. Only 10% of the cancer patients had combined daily intake levels corresponding to the threshold of sensitivity of the micronucleus test as defined by previous studies. The mean frequency of micronucleated buccal cells was 0.26% in the cancer patients and 0.13% in the non-cancerous subjects. All non-cancerous patients presented a negative test. Only 5% of the cancer patients presented a micronucleated cell frequency above 1% and could thus be considered as positive. It thus appears that the micronucleus test was not significantly positive in our population of 38 cancer patients.