Gait analysis of a patient after femoral nerve and malignant soft tissue tumor resections: a case report.

BACKGROUND: Malignant femoral soft tissue tumors are occasionally resected together with the femoral nerves, but this can cause loss of knee extensor muscle activity. To the best of our knowledge, no previous reports have detailed the gait analysis of such cases in combination with electromyography. Herein, we report the gait analysis of a patient who underwent left groin synovial sarcoma and left femoral nerve resection 12 years ago. CASE PRESENTATION: We analyzed the gait of a 38-year-old man who was able to walk unaided after the resection of a synovial sarcoma in the left groin together with the ipsilateral femoral nerve. The muscle activities of the affected medial (MH) and lateral hamstrings (LH), and lateral heads of the gastrocnemius (GL) were increased during 50-75% of the stance phase. The hip flexion angle of the affected limb was smaller, and the ankle plantar flexion angle of the affected limb was larger than that of the non-affected limb. This means that in the affected limb, the hip and ankle angles were adjusted to prevent knee collapse, and the MH, LH, and GL muscles contributed in the mid- and late-stance phases. Moreover, we found that the hamstring and gastrocnemius of the affected limb worked together to keep the ipsilateral knee extended in the mid-stance phase and slightly flexed in the late-stance phase. CONCLUSIONS: Patients capable of walking after femoral nerve resection may control their hamstrings and gastrocnemius muscles collaboratively to prevent ipsilateral knee collapse in the mid- and late-stance phases.

MicroRNA-329-3p inhibits the Wnt/ß-catenin pathway and proliferation of osteosarcoma cells by targeting transcription factor 7-like 1.

An important factor in the emergence and progression of osteosarcoma (OS) is the dysregulated expression of microRNAs (miRNAs). Transcription factor 7-like 1 (TCF7L1), a member of the T cell factor/lymphoid enhancer factor (TCF/LEF) transcription factor family, interacts with the Wnt signaling pathway regulator ß-catenin and acts as a DNA-specific binding protein. This study sought to elucidate the impact of the interaction between miR-329-3p and TCF7L1 on the growth and apoptosis of OS and analyze the regulatory expression relationship between miRNA and mRNA in osteosarcoma cells using a variety of approaches. MiR329-3p was significantly downregulated, while TCF7L1 was considerably up-regulated in all examined OS cell lines. Additionally, a clinical comparison study was performed using the TCGA database. Subsequently, the regulatory relationship between miR-329-3p and TCF7L1 on the proliferation and apoptosis of OS cells was verified through in vitro and in vivo experiments. When miR-329-3p was transfected into the OS cell line, the expression of TCF7L1 decreased, the proliferation of OS cells was inhibited, the cytoskeleton disintegrated, and the nucleus condensed to form apoptotic bodies. The expression of proteins that indicate apoptosis increased simultaneously. The cell cycle was arrested in the G0/G1 phase, and the G1/S transition was blocked. The introduction of miR-329-3p also inhibited downstream Cyclin D1 of the Wnt pathway. Xenograft experiments indicated that the overexpression of miR-329-3p significantly inhibited the growth of OS xenografts in nude mice, and the expression of TCF7L1 and c-Myc in tumor tissues decreased. MiR-329-3p was significantly reduced in OS cells and played a suppressive role in tumorigenesis and proliferation by targeting TCF7L1 both in vitro and in vivo. Osteosarcoma cell cycle arrest and pathway inhibition were observed upon the regulation of TCF7L1 by miR-329-3p. Summarizing these results, it can be inferred that miR-329-3p exerts anticancer effects in osteosarcoma by inhibiting TCF7L1.

Sequential treatments with TRK inhibitors in a patient with NTRK fusion-positive sarcoma: A case report.

RATIONALE: Precision medicine and tumor-agnostic treatment strategies have recently been promoted for clinical use. One of the most successful treatments in patients with neurotrophic tyrosine receptor kinase (NTRK) fusion-positive tumors is targeting the tropomyosin receptor kinase (TRK) with an inhibitor. The TRK inhibitors, larotrectinib, and entrectinib, have been approved in many countries. Nevertheless, the most effective administration regimen for these TRK inhibitors is uncertain. To date, no reports have shown the efficacy of sequential treatment with larotrectinib and entrectinib in patients with NTRK fusion-positive tumors. In this report, we present a patient with NTRK fusion-positive sarcoma arising from the anterior mediastinum, with tumor progression after 4 months of entrectinib use. The patient took larotrectinib subsequently and maintained disease control for more than 21 months. PATIENT CONCERNS: A 48-year-old female visited a physician because she experienced difficulty in breathing and chest and back pain with no obvious cause 2 months ago. Computed tomography (CT)-guided biopsy was performed at a district general hospital, and histopathological examination revealed a small round cell tumor. She was referred to our hospital, and a second CT-guided biopsy was performed to confirm the pathological diagnosis. Considering the results of the histopathological examination, Ewing sarcoma was suspected, but a specific fusion gene was not detected due to poor quality specimens. DIAGNOSES: After 3 regimens of cytotoxic chemotherapy, biopsy was repeated, and specimens were analyzed using next-generation sequencing. The PHF20-NTRK1 fusion gene was detected, and the tumor was finally diagnosed as an NTRK fusion-positive sarcoma. INTERVENTIONS: She was administered the TRK inhibitor entrectinib, but the tumor started to grow after 4 months of medication, and she stopped taking entrectinib. After 1 cycle of cytotoxic chemotherapy, another TRK inhibitor, larotrectinib, was administered. OUTCOMES: Her stable disease was maintained for more than 21 months. Here, we have shown that sequential administration of both drugs can be effective. LESSONS: In the treatment of NTRK fusion-positive tumors, there are cases in which 2 approved first-generation TRK inhibitors can be used sequentially.

Expression of Glial-Cell-Line-Derived Neurotrophic Factor Family Ligands in Human Intervertebral Discs.

Glial-cell-line-derived neurotrophic factor (GDNF) family ligands (GFLs) contribute to the sensitization of primary afferents and are involved in the pathogenesis of inflammatory pain. The purpose of this preliminary study was to examine the expression of other GFLs (neurturin (NRTN), artemin (ARTN), persephin (PSPN)) and receptors in human IVD cells and tissues exhibiting early and advanced stages of degeneration. Human IVD cells were cultured as a monolayer after isolation from the nucleus pulposus (NP) and anulus fibrosus (AF) tissues. The mRNA expression of NRTN, ARTN, PSPN, and their receptors (GFRA2-GFRA4) was quantified using real-time PCR. Protein expression was evaluated using immunohistochemistry and Western blotting. The expression of NRTN, ARTN, PSPN, and their co-receptors (GFRA2-GFRA4) was identified in human IVD cells at both mRNA and protein levels. A trend was noted wherein the mRNA expression of ARTN, PSPN, and GFRA2 was upregulated by IL-1ß treatment in a dose-dependent manner. The percentages of immunopositive cells in the advanced degenerate stage of ARTN, PSPN, and GFRA2 were significantly higher than those in the early degenerate stage. Their expression was enhanced in advanced tissue degeneration, which suggests that GFLs (ARTN and PSPN) may be involved in the pathogenesis of discogenic pain.

Tumor-suppressive microRNA-152 inhibits the proliferation of Ewing's sarcoma cells by targeting CDK5R1.

We elucidated the mechanism through which the reduced expression of miR-152 leads to the overexpression of its target cyclin-dependent kinase-5 activator 1 (CDK5R1) in Ewing's sarcoma (ES) cells and the role of this mechanism in the proliferation of ES cells. To explore possible oncogenic factors in ES, we conducted microarray-based investigation and profiled the changes in miRNA expression and their effects on downstream mRNAs in five ES cell lines and human mesenchymal stem cells (hMSCs). miR-152 was significantly downregulated, while cyclin-dependent kinase-5 activator 1 (CDK5R1) expression was significantly upregulated in all tested ES cells as compared to hMSCs. The overexpression of CDK5R1 led to the activation of CDK5, enabling the phosphorylation of retinoblastoma protein and persistent overexpression of CCNE. Moreover, miR-152 suppressed cell proliferation via cell cycle retardation, and its upregulation reduced tumor size and CCNE expression in tumor tissues. The overexpression of cyclin E (CCNE) has been detected in ES cells, but the detailed mechanisms have not been previously elucidated. These findings identify the miR152-CDK5R1 signaling axis as a critical mechanism for tumorigenesis that may serve as a new therapeutic target in Ewing's sarcoma. We believe that our results will aid in the development of effective treatment strategies for patients with ES.

The anti-oncogenic effect of 17-DMAG via the inactivation of HSP90 and MET pathway in osteosarcoma cells.

Heat shock protein (HSP) 90 plays a crucial role in correcting the misfolded three-dimensional structure of proteins, assisting them in folding into proper conformations. HSP90 is critical in maintaining the normal functions of various proteins within cells, as essential factors for cellular homeostasis. Contrastingly, HSP90 simultaneously supports the maturation of cancer-related proteins, including mesenchymal epithelial transition factor (MET) within tumor cells. All osteosarcoma cell lines had elevated MET expression in the cDNA array in our possession. MET, a tyrosine kinase receptor, promotes proliferation and an anti-apoptotic state through the activation of the MET pathway constructed by HSP90. In this study, we treated osteosarcoma cells with an HSP90 inhibitor, 17-demethoxygeldanamycin hydrochloride (17-DMAG), and assessed the changes in the MET signaling pathway and also the antitumor effect of the drug. The cell cycle in osteosarcoma cells administered 17-DMAG was found to be halted at the G2/M phase. Additionally, treatment with 17-DMAG inhibited cell proliferation and induced apoptosis. Inhibition of tumor cell proliferation was also observed in an in vivo model system, mice that were treated with 17-DMAG. Based on the results of this study, we were able to confirm that 17-DMAG promotes inhibition of osteosarcoma cell proliferation and induction of apoptosis by inhibition of MET, a protein highly expressed in osteosarcoma cells. This approach may be useful for the establishment of a new treatment strategy for patients resistant to the standard treatment for osteosarcoma.

Evaluation of the two-point calibration bromocresol green method, showing reduced deviation from the bromocresol purple method in sera from patients with hypoalbuminemia.

BACKGROUND: The bromocresol green (BCG) and bromocresol purple (BCP) methods are widely used for albumin measurements in routine testing, but the BCG method is known to react with globulin fractions and to have low specificity for albumin. We evaluated a calibration method using different concentrations of human serum albumin standards (two-point calibration BCG method) with the aim of reducing the effect of globulin fractions on the BCG method in patients with hypoalbuminemia. METHOD: In the two-point calibration BCG method, two concentrations of standard solutions and their calibration values are set based on the difference in albumin concentrations measured by the BCG method (BCG-HSA method) and the modified BCP (modified BCP-HSA method) calibrated with human serum albumin standard solution (HSA). Albumin concentrations were measured in 136 patient serum samples (healthy group: 52, hypoalbuminemic group: 84) by the two-point calibrated BCG method and compared with those obtained using the modified BCP-HSA method. RESULTS: The mean albumin concentrations obtained using the two-point calibrated BCG and modified BCP-HSA methods were 39.18 ± 3.42 g/L and 39.37 ± 3.14 g/L (healthy group) and 26.20 ± 6.23 g/L and 26.23 ± 5.67 g/L (hypoalbuminemia group), respectively. The results of the two-point calibration BCG method were in a close agreement over the entire concentration range tested compared to the modified BCP-HSA method. CONCLUSIONS: Based on these results, this calibration method reduces the influence of the globulin fraction on the BCG method. In the hypoalbuminemic group, the calibration method was shown to provide results consistent with the BCP method, which is highly specific for albumin.

Relationship between K-line distance and surgical outcome in cases of laminoplasty for cervical ossification of the posterior longitudinal ligament.

Although previous studies indicate that changes in cervical alignment after laminoplasty and dynamic factors influence surgical outcomes of cervical ossification of the posterior longitudinal ligament (OPLL), the relationship between the surgical outcomes, the distance between the kyphosis-line (K-line) and OPLL, and dynamic factors have not yet been quantitatively evaluated. The purpose of the present study was to analyze the relationship between ΔK-line distance and surgical outcomes in cases of laminoplasty for OPLL of the cervical spine. We retrospectively reviewed 46 consecutive patients (33 men and 13 women) with cervical OPLL who underwent laminoplasty. "K-line distance" was measured as the minimum interval between the K-line and OPLL on lateral radiographs. The following factors were analyzed: K-line distance in neutral, flexion, and extension neck positions, ΔK-line distance, preoperative C2-7 range of motion (ROM), preoperative segmental ROM, preoperative C2-7 lordotic angle, occupying ratio of the OPLL, disease duration, preoperative and postoperative Japanese Orthopaedic Association (JOA) score, and recovery rate. Patients were divided into flexion K-line (+) and flexion K-line (-) groups. We then analyzed the influence of the K-line distance on surgical outcomes and conducted multivariate analysis to analyze the factors affecting surgical outcomes. The JOA score recovery rate in the flexion K-line (-) group was significantly lower than that in the flexion K-line (+) group (P = .024). The ΔK-line distance was significantly negatively correlated with the JOA score recovery rate (r = -0.531, P < .001). Additionally, multivariate analysis showed that ΔK-line distance (OR = -2.143, P = .015) was negatively correlated with the JOA score recovery rate. The ΔK-line distance is considered useful for the quantitative evaluation of dynamic factors and static compression factors due to OPLL through the measurement of dynamic radiographic images.

Surrogacy analysis of intermediate end-points for overall survival in randomized controlled trials of rhabdomyosarcoma.

Treatment of malignant tumors, such as rhabdomyosarcoma (RMS), can improve overall survival (OS). It is time-consuming and expensive for patients to obtain benefits from randomized controlled trials (RCTs) with OS as the primary end-point. Therefore, another surrogate end-point is necessary; however, there is no report on the surrogacy analysis of RMS. In this study, we performed a systematic review of RCTs, involving patients with newly diagnosed RMS, and 11 RCTs were identified. We performed a meta-analysis to assess the surrogacy of intermediate end-points for OS. The correlations between surrogate end-points and OS were investigated using Spearman's rank correlation coefficient (ρ). The coefficient of determination (R2) was employed to measure the strength of the association. A total of 5183 patients were randomly allocated to 34 treatment groups. A marginal correlation (R2 = 0.281, ρ = 0.445) between the hazard ratios (HRs) for event-free survival (EFS) and OS was observed. In patients with localized RMS, the EFS HR had a weaker correlation with OS HR in the sensitivity analysis than that in the primary analysis. Overall, the surrogacy of EFS for OS cannot be confirmed.

Analysis of rotational deformity correction by lateral lumbar interbody fusion with two-staged anterior-posterior combined corrective fusion surgery for adult degenerative kyphoscoliosis.

The present study is retrospective analysis of consecutively collected data. Lateral lumber interbody fusion (LLIF) is widely used in cases of adult spinal deformities. However, the corrective effects of LLIF cage insertion on the vertebral rotation deformity in the axial plane and the individual effects of LLIF and direct vertebral rotation (DVR) on rotational correction are unclear. To individually examine the corrective effects of LLIF and posterior corrective fusion surgery with direct DVR on vertebral rotation deformities in adult degenerative kyphoscoliosis. We analyzed 21 patients (5 males and 16 females) who underwent two-staged anterior-posterior combined corrective fusion surgery for adult degenerative kyphoscoliosis. Surgical time, blood loss, facet joint osteoarthritis (OA) grade, disc degeneration, cage height, vertebral rotational angle, and various X-ray parameters were investigated as evaluation items. The X-ray parameters showed significant postoperative improvements. The mean vertebral rotation angle was 6.4°â€…±â€…5.2° preoperatively, 3.5°â€…±â€…3.3° after LLIF (P = .014, vs preoperative), and 1.6°â€…±â€…1.7° after posterior corrective fusion surgery with DVR (P = .011, vs preoperative). Correlation analysis between the vertebral rotation angle and various measured values revealed that the vertebral rotation angle after LLIF was correlated with the cage height (r = -0.46, P = .032). The vertebral rotation angle after DVR was correlated with the facet joint OA grade (r = -0.49, P = .018) and the wedge angle after posterior corrective fusion surgery with DVR (R = 0.57, P = .006). We conclude that the effects of rotational deformity correction with LLIF cage insertion and additional posterior corrective fixation with DVR can be useful for correcting vertebral rotation deformities.

The effects of romosozumab combined with active vitamin D3 on fracture healing in ovariectomized rats.

BACKGROUND: In this study, we investigated the potential acceleration of fracture healing and bone mineral density-increasing effects of romosozumab and active vitamin D3 combination therapy for fractures in ovariectomized rats. METHODS: Ovariectomy was performed on 40 24-week-old female Sprague-Dawley rats. After 8 weeks, the rats were subjected to periosteum removal and osteotomy of the femoral shaft followed by osteosynthesis with intramedullary nailing to create fracture models. The rats were then divided into four groups: C group (control), R group (receiving romosozumab at 25 mg/kg once a month via subcutaneous injection), VD group (receiving active vitamin D3 at 0.2 µg/kg twice a week via subcutaneous injection), and R + VD group. Further, 10 rats were included in a sham group. At 10 weeks after the intervention, both femurs were removed and blood samples were collected from all rats. Soft X-ray imaging was used to evaluate bone union, and microcomputed tomography (micro-CT) was used for bone morphometric evaluation. Toluidine blue staining was used for the histopathological evaluation of the undecalcified specimens, and bone turnover marker levels were measured using enzyme-linked immunosorbent assay. RESULTS: Bone morphometry analysis via micro-CT revealed increased mineral density of the trabecular bone in the R + VD group femurs, demonstrating the effectiveness of romosozumab plus active vitamin D3 combination therapy. However, there were no differences in bone union evaluated using soft X-ray imaging, indicating no acceleration of fracture healing. CONCLUSIONS: Although romosozumab and active vitamin D3 combination therapy increased trabecular bone volume, there was no evidence on its ability to accelerate fracture healing.

Analysis of treatment effect with teriparatide on device-related vertebral osteopenia after lumbar spinal interbody fusion using Hounsfield unit values: A retrospective cohort study.

The aim of the present study was to investigate the effect of teriparatide on device-related vertebral osteopenia after single lumbar spinal interbody fusion and compare osteopenia in fused and nonfused spinal segments using Hounsfield unit (HU) values. The present study was a retrospective cohort study. We reviewed 68 consecutive patients (28 men and 40 women) who underwent single-segment (L4-5) transforaminal lumbar interbody fusion with cage and pedicle screw fixation. The patients were divided into 2 groups according to whether they were treated with teriparatide (teriparatide and nonmedication groups). The primary outcome measure was HU values measured on computed tomography images from each L1 to S1 vertebral body12-month postoperatively. Secondary outcome measures were femoral neck bone mineral density (BMD), T-score, osseous union, and clinical outcomes using the Japanese Orthopedic Association scoring system 12-month postoperatively. There were significant decreases in HU values of lumbar vertebral bodies at all levels and BMD and T-score values obtained using dual-energy X-ray absorptiometry of the femur between preoperative and postoperative 12-month computed tomography in the nonmedication group (P < .05). On the other hand, there were no significant differences between properative and postoperative 12-month HU values of each lumbar vertebral body and BMD values of the femur in the teriparatide group. Osseous fusion scores in the teriparatide group were significantly better than those in the nonmedication group. There were no significant differences in postoperative Japanese Orthopedic Association scores between the 2 groups. Administration of teriparatide during the perioperative period may prevent bone loss associated with spinal fusion surgery.

Patellar dislocation following distal femoral replacement after extra-articular knee resection for bone sarcoma: A case report.

BACKGROUND: For the treatment of bone sarcoma in the distal femur, wide-margin resection and knee reconstruction with tumor endoprosthesis are standard therapies. Extra-articular knee resection is required in cases of tumor invasion of the knee joint; however, the incidence of complications, such as aseptic loosening, prosthesis infection, and implant failure, is higher than that following intra-articular knee resection. To the best of our knowledge, there are three reports of patellar dislocations after replacement of a tumor endoprosthesis. CASE SUMMARY: A 36-year-old man with no significant past medical history was admitted to our institution with continuous pain in his left knee for 4 mo. An open biopsy was performed, and the patient was diagnosed with a left distal femoral malignant bone tumor. Extra-articular knee resection and knee reconstruction with a tumor endoprosthesis were performed. Although the alignment of the tumor prosthesis was acceptable, knee instability was noticed postoperatively. The axial radiographic view of the patellar and computed tomography showed lateral patellar dislocation at 4 wk postoperatively. The patient had to undergo a lateral release and proximal realignment. He could perform his daily activities at 9 mo postoperatively. Radiography revealed no patellar re-dislocation. CONCLUSION: Proximal realignment may be considered during primary surgery if there is an imbalance in the forces controlling the patellar tracking.

Analysis of the signal cross talk via CCL26 in the tumor microenvironment in osteosarcoma.

Interaction with surrounding healthy cells plays a major role in the growth and metastasis of osteosarcoma. In this study, we hypothesized that humoral factors, which do not require direct contact with cells, are involved in the interaction between osteosarcoma and the surrounding cells. We identified the humoral factor involved in the association between tumor cells and surrounding normal cells using a co-culture model and investigated the significance of our findings. When human osteosarcoma cells (MG63) and human mesenchymal stem cells (hMSCs) were co-cultured and comprehensively analyzed for changes in each culture group, we found that the expression of chemokine (CC motif) ligand 26 (CCL26) was significantly enhanced. We also analyzed the changes in cell proliferation in co-culture, enhanced interaction with administration of recombinant CCL26 (rCCL26), reduced interaction with administration of anti-CCL26 antibodies, changes in invasive and metastatic abilities. CCL26 levels, motility, and invasive capability increased in the co-culture group and the group with added rCCL26, compared to the corresponding values in the MG63 single culture group. In the group with added CCL26 neutralizing antibodies, CCL26 level decreased in both the single and co-culture groups, and motility and invasive ability were also reduced. In a nude mice lung metastasis model, the number of lung metastases increased in the co-culture group and the group with added rCCL26, whereas the number of tumors were suppressed in the group with added neutralizing antibodies compared to those in the MG63 alone. This study identified a possible mechanism by which osteosarcoma cells altered the properties of normal cells to favorably change the microenvironment proximal to tumors and to promote distant metastasis.

Sensitive quantification of free pazopanib using ultra-high performance liquid chromatography coupled to tandem mass spectrometry and assessment of clinical application.

Pazopanib is widely used to treat renal cell carcinomas and soft tissue tumors in Japan. Although several reports demonstrated the usefulness of therapeutic drug monitoring (TDM) of pazopanib, those studies measured only total pazopanib concentration. For drugs with high protein binding rates such as pazopanib, measuring free concentrations may be clinically more useful than measuring total concentrations. In this study, we aimed to develop a high-throughput method for quantification of free pazopanib in human plasma using ultra-high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS). Free pazopanib was separated by ultrafiltration. After a simple solid-phase extraction step using a 96-well plate, pazopanib was analyzed by UHPLC-MS/MS in positive electrospray ionization mode. The novel method fulfilled the requirements of the US Food and Drug Administration guidelines for assay validation, and the lower limit of quantification was 0.05 ng/mL. The calibration curve was linear over the concentration range of 0.05-50 ng/mL. The average recovery rate was 66.9 ± 2.1% (mean ± SD). The precision was below 7.02%, and accuracy was within 10.60% across all quality control levels. Matrix effect varied between 44.4% and 60.4%. This assay was successfully applied to measure trough free pazopanib concentrations in three patients treated with pazopanib for soft tissue tumors. We succeeded to develop a novel high-throughput UHPLC-MS/MS method for quantification of free pazopanib in human plasma. This method can be applied to TDM for patients receiving pazopanib in the clinical setting.

Gait analysis of a patient who underwent complete resection of the patella and quadriceps femoris for soft tissue sarcoma.

The knee extension mechanism including the quadriceps femoris muscles and patella plays a crucial role in the stance phase of a normal gait cycle. We performed gait analysis of a patient who had undergone complete resection of the knee extension mechanism. An 8-month-old boy developed infantile fibrosarcoma of the right knee and underwent resection of the quadriceps femoris muscles, patella, and patellar tendon. The gait analysis performed at 8 years of age demonstrated that he could maintain the knee joint extension position during the stance phase. Increased muscle activities in the hamstring and gastrocnemius were observed. The results suggest that the hamstring and gastrocnemius muscles might play a role in maintaining the knee extension position during the stance phase. We suggest the importance of reinforcing these muscles in rehabilitation for patients who lost the knee extension mechanism.

Primary clear cell sarcoma of the femur: a unique case with RT-PCR and direct sequencing confirmation of EWSR1/ATF1 fusion gene.

BACKGROUND: It is very rare for clear cell sarcomas (CCS) to arise in the bone. During diagnosis, it is important to distinguish primary CCS of bone from bone metastasis of melanoma because this difference fundamentally changes the therapeutic options. Recently, characteristic fusion genes of CCS have been detected using reverse transcription polymerase chain reaction (RT-PCR) or direct sequencing which allowed to distinguish CCS from melanoma. However, there was no study applying these analyses with positive results. In this case, we describe the use of fusion gene analysis to diagnose a primary CCS of the bone. CASE PRESENTATION: A 36-year-old male presented with a four-months history of left knee pain. Magnetic resonance imaging showed a lesion in the left femoral medial epicondyle. Histological examination of the biopsy specimen revealed proliferating oval or rounded cells. These cells had clear cytoplasm arranged in fascicles or compact nests with frequent deposits of brown pigment. Furthermore, immunohistochemistry analysis revealed that tumor cells were positive for S-100 protein, HMB-45, Melan-A, and SOX10. It stained negative for CD34 and BRAF v600e. Conclusively, detection of the EWSR1/ATF1 fusion gene using RT-PCR and direct sequencing confirmed that the lesion was a primary CCS of the bone. Wide-margin resection and reconstruction with a tumor endoprosthesis were performed. CONCLUSIONS: Herein, we diagnosed a rare case of primary CCS of the bone by detecting EWSR1/ATF1 fusion gene using RT-PCR and direct sequencing. Since fluorescence-in situ hybridization (FISH) and RT-PCR could show false positive by mainly due to technical problems, it is better to perform direct sequencing to confidently diagnose the tumor as a primary CCS especially at very rare site such as bone.

Development of a High-Throughput Quantification Method for Pazopanib Using Ultra-Performance Liquid Chromatography-Tandem Mass Spectrometry and Its Clinical Application in Patients With Soft Tissue Tumors.

BACKGROUND: Pazopanib is widely used to treat renal cell carcinomas and soft tissue tumors in Japan. Pazopanib has significant therapeutic efficacy but it is associated with frequent severe adverse effects. Therapeutic drug monitoring (TDM) may help to prevent adverse effects. A more convenient and rapid pazopanib assay is desirable for the application of TDM in clinical settings. In this study, the authors developed a high-throughput method for quantifying pazopanib in human plasma using ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). METHODS: After a simple solid-phase extraction step using a 96-well plate, pazopanib was analyzed by UHPLC-MS/MS in the positive electrospray ionization mode. RESULTS: The novel method fulfilled the requirements of the US Food and Drug Administration and the European Medicines Agency guidelines for assay validation, and the lower limit of quantification was 0.5 mcg/mL. The calibration curves were linear over the concentration range of 0.5-100 mcg/mL. The average recovery rate was 102.0% ± 3.9% (mean ± SD). The precision was below 5.0%, and the accuracy was within 12.0% for all quality control levels. Matrix effect varied between 90.9% and 97.1%. This assay was successfully applied to TDM of pazopanib trough concentrations in 3 patients treated with the drug for soft tissue tumors. CONCLUSIONS: The authors succeeded in developing a novel high-throughput UHPLC-MS/MS method for quantifying pazopanib in human plasma. This method can be applied to TDM of patients receiving pazopanib in clinical settings.

Osteomyelitis of the Distal Phalanx of the Thumb due to Parvimonas micra and Fusobacterium nucleatum: A Case Report.

We herein report a case of osteomyelitis of the distal phalanx of the thumb of a 55-year-old man caused by Parvimonas micra and Fusobacterium nucleatum. Osteomyelitis often occurs in long bones and rarely occurs in the bones of the fingers. In addition, osteomyelitis of the finger frequently occurs after trauma or surgery, and blood-borne infection is very rare. P. micra and F. nucleatum, normal flora of the oral cavity, are very rare pathogenic bacteria of osteomyelitis except in periodontal disease, and there are no previous reports regarding the occurrence of osteomyelitis due to P. micra and F. nucleatum in the finger bones.

A meta-analytic evaluation of the correlation between event-free survival and overall survival in randomized controlled trials of newly diagnosed Ewing sarcoma.

BACKGROUND: In randomized controlled trials (RCTs) of adjuvant treatment for malignant tumors, event-free survival (EFS) is considered the most acceptable surrogate for overall survival (OS). However, even though EFS has repeatedly been selected as a primary endpoint in RCTs of Ewing sarcoma (ES), the surrogacy of EFS for OS has not been investigated. This study aimed to evaluate the correlation between EFS and OS in RCTs of chemotherapy for newly diagnosed ES using a meta-analytic approach. METHODS: We identified seven RCTs of newly diagnosed ES through a systematic review, and a meta-analysis was performed to evaluate the efficacy and adverse events associated with chemotherapy for previously untreated ES. The correlation between EFS and OS was investigated using weighted linear regression analysis and Spearman rank correlation coefficients (ρ). The strength of the correlation was evaluated using the coefficient of determination (R2). RESULTS: A total of 3612 patients were randomly assigned to 17 treatment arms in the eligible RCTs. The meta-analysis revealed that the hazard ratios for OS and EFS showed significantly better results in the experimental treatment groups with increasing toxicities. The correlation between the hazard ratios for EFS and OS was good (R2 = 0.747, ρ = 0.683), and the correlation tended to be more favorable in cases of localized ES (R2 = 0.818, ρ = 0.929). CONCLUSIONS: Overall, the trial-level correlation between EFS and OS was good for newly diagnosed ES and was very good in cases of localized disease. EFS may be a useful endpoint in RCTs of ES chemotherapy, and it is worth verifying using individual patient data.