Tranexamic Acid

After centrifugation, cell pellets from the enriched and column flow-through fractions were resuspended to 100 µl and 2 ml, respectively, with sorter buffer, and 5 µl was removed for cell counting. Cell suspensions were incubated with surface antibodies for 25 min on ice and washed with sorter buffer. Surface antibodies, used in various combinations, were as follows: FITC-, AF647-, or Biotin-labeled GL7 (BD or eBioscience); AF700- or Pacific blue–labeled anti-CD38 (eBioscience or BioLegend), V500- or eF450-labeled anti-B220 (BD or eBioscience); AF700- or eF605NC-labeled anti-CD19 (eBioscience); eF450-labeled anti-CD21/35 (eBioscience); FITC-, PE-Cy7-, or Biotin-labeled anti-CD23 (eBioscience); eF605NC-labeled anti-CD24 (eBioscience); Biotin-labeled anti-CD40 (eBioscience); FITC-labeled anti-CD43 (clone S7; BD); AF700-labeled anti-CD44 (eBioscience); PerCP-Cy5.5-labeled anti-CD80 (BioLegend); APC-labeled anti-CD86 (eBioscience); eF605NC-labeled anti-CD93 (clone AA4.1; eBioscience), FITC-labeled anti-CD95 (eBioscience); FITC-labeled Igβ (CD79b; BioLegend); AMCA- or PE-Cy7–labeled anti-IgM (Jackson ImmunoResearch Laboratories or eBioscience); PerCP-Cy5.5– or eF450-labeled anti-IgD (BioLegend or eBioscience); APC-labeled anti-IgG1 (BD); and APC-eF780–labeled anti-CD11c, anti-CD4, anti-CD8, anti-Thy1.2, anti-F4/80, and anti-Gr-1 (eBioscience). In some experiments where biotin-labeled antibodies were used, cells were then stained with FITC-, eF605NC or PE-Cy7-labeled streptavidin (eBioscience) for 15 min on ice and washed with sorter buffer. In experiments where cell fixation was not required, cells were resuspended in 0.02 µg/ml DAPI (Sigma-Aldrich) and DAPI⁺ cells excluded from analysis.
For intracellular staining, pelleted samples were resuspended in 250 µl Cytofix/Cytoperm (BD) for 25 min on ice and washed in permeabilization buffer (BD) before labeling with AF350-conjugated goat anti–mouse Ig H+L (Jackson ImmunoResearch Laboratories) for 30 min on ice and being washed with permeabilization buffer.
Flow cytometry was performed on a 4-laser (355 nm, 405 nm, 488 nm, and 633 nm) or 5-laser (355 nm, 405 nm, 488 nm, 561 nm, and d640 nm) LSR II device (BD) and analyzed with FlowJo software (Tree Star). Fluorescent AccuCheck counting beads (Invitrogen) were used to calculate total numbers of live lymphocytes in the column-bound and flow-through suspensions, as previously described (Pape et al., 2011 (link)). Samples collected after OVA injection or KRN adoptive transfer often had a significant number of tetramer-specific B cells that were not retained by the column. When this occurred, the number of tetramer-specific cells in the column flow through fraction was included in the total number of cells.

Postoperative management included: Conventional antibiotics were applied intraoperatively to prevent infection, and antibacterial drugs were applied prophylactically within 24 hours postoperatively. Tranexamic acid 1 was given intravenously 3 hours postoperatively. Subcutaneous anticoagulation with 4100 U of low-molecular heparin was started 10 hours after surgery, qd × 5 days. After discharge, the patient was given oral rivaroxaban 10 mg, qd × 14 days. Ice packs were applied intermittently for 48 hours after surgery, and the dressing was changed every other day. The incision was removed at 12 to 14 days postoperatively with outpatient review. The patient started normal weight-bearing walking with the aid of a walker 1 day after surgery THA. Patients were instructed to actively flex and extend the knee joint, perform ankle pump exercises and quadriceps isometric contraction exercises as well as passive exercises 1 day after surgery.
Validated updated version of diagnostic criteria for PJI in 2018 are as follows^{[13 (link)]} (based on the diagnostic criteria of PJI proposed by the Musculoskeletal infection society in 2011):
Two positive cultures or the presence of a sinus tract were considered major criteria and diagnostic of PJI. The calculated weights of an elevated serum CRP (>1 mg/dL), D-dimer(>860ng/mL) and ESR (>30mm/hour) were 2, 2 and 1 points, respectively. Elevated synovial fluid WBC count (>3000 cells/µL), alpha-defensin (signal-to cutoff ratio > 1), LE (++), PMN% (>80%) and synovial CRP (>6.9mg/L) received 3, 3, 3, 2 and 1 points, respectively. Patients with an aggregate score of greater than or equal to 6 were considered infected while a score between 2 and 5, required the inclusion of intraoperative findings for confirming or refuting the diagnosis. Intraoperative findings of positive histology, purulence and single positive culture were assigned 3, 3, and 2 points, respectively. Combined with the preoperative score, a total of greater than or equal to 6 was considered infected, a score between 4 and 5 was inconclusive, and a score of 3 or less was not infected.