Molecular characterization of ferroptosis in soft tissue sarcoma constructs a prognostic and immunotherapeutic signature through experimental and bioinformatics analyses.

Ferroptosis regulators have been found to affect tumor progression. However, studies focusing on ferroptosis and soft tissue sarcoma (STS) are rare. Somatic mutation, copy number variation, reverse transcription-quantitative polymerase chain reaction (RT-qPCR) analysis, consensus clustering, differentially expressed genes analysis (DEGs), principal component analysis (PCA) and gene set enrichment analysis (GSEA) were used to identify and explore different ferroptosis modifications in STS. A nomogram was constructed to predict the prognosis of STS. Moreover, three immunotherapy datasets were used to assess the Fescore. Western blotting, siRNA transfection, EdU assay and reactive oxygen species (ROS) measurement were performed. 16 prognostic ferroptosis regulators were screened and significant differences were observed in somatic mutation, copy number variation (CNV) and RT-qPCR among these ferroptosis regulators. 2 different ferroptosis modification patterns were found (Fe cluster A and B). Fe cluster A with higher Fescore was correlated with p53 pathway and had better prognosis of STS (p = 0.002) while Fe cluster B with lower Fescore was correlated with angiogenesis and MYC pathway and showed a poorer outcome. Besides, the nomogram effectively predicted the outcome of STS and the Fescore could also well predict the prognosis of other 16 tumors and immunotherapy response. Downregulation of LOX also inhibited growth and increased ROS production in sarcoma cells. The molecular characterization of ferroptosis regulators in STS was explored and an Fescore was constructed. The Fescore quantified ferroptosis modification in STS patients and effectively predicted the prognosis of a variety of tumors, providing novel insights for precision medicine.

Molecular Characteristics of m6A Regulators and Tumor Microenvironment Infiltration in Soft Tissue Sarcoma: A Gene-Based Study.

Background: N6-methyladenosine (m6A) methylation played a key role in tumor growth. However, the relationship between m6A and soft tissue sarcoma (STS) was still unclear. Methods: The characterization and patterns of m6A modification in STS (TCGA-SARC and GSE17674) were analyzed comprehensively through bioinformatics and real-time polymerase chain reaction (RT-PCR). The effects of different m6A modification patterns on prognosis and immune infiltration of STS were further explored. Differentially expressed gene (DEG) analysis was performed. Moreover, an m6Ascore was constructed by principal component analysis (PCA). In addition, two immunotherapy datasets (IMvigor210 and GSE78220) and a sarcoma dataset (GSE17618) were used to evaluate the m6Ascore. Results: Huge differences were found in somatic mutation, CNV, and expression of 25 m6A regulators in STS. Two modification patterns (A and B) in STS were further identified and the m6A cluster A showed a better clinical outcome with a lower immune/stromal score compared with the m6A cluster B (p < 0.050).In addition to , most STS samples from m6A cluster A showed a high m6Ascore, which was related to mismatch repair and a better prognosis of STS (p < 0.001). In contrast, the m6A cluster B, characterized by a low m6Ascore, was related to the MYC signaling pathway, which led to a poor prognosis of STS. A high m6Ascore also contributed to a better outcome of PD-1/PD-L1 blockade immunotherapy. Conclusion: The modification patterns of 25 m6A regulators in the STS microenvironment were explored comprehensively. The novel m6Ascore effectively predicted the characteristics of the tumor microenvironment (TME) and outcome in STS and provided novel insights for future immunotherapy.

Incomplete protective effects of minocycline on traumatic brain injury in rats and mice / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To evaluate protective effect of minocycline,a semisynthetic tetracycline derivative on different traumatic brain injuries in rats and mice.</p><p><b>METHODS</b>The opened brain trauma was induced in rats and the closed head injury and cold brain injury were induced in mice. In 3 brain trauma models, minocycline (45 mg/kg, ip) was administered twice daily for 2 d before the operation, at 30 min before and 1 h after the operation, and once daily for 2 d following the operation (totally 8 doses in 5 d). After the operation, the behavioral alteration was observed daily, lesion area and survival neuron density were measured at the end of the experiments (14 d after the injuries).</p><p><b>RESULT</b>For rat opened traumatic injury, minocycline promoted the recovery of hindlimb motor activity (inclined board angle), but did not alter other indexes. For mouse closed head traumatic injury, minocycline reduced the neuron loss, but did not improve behavioral dysfunction. For mouse cold injury-induced trauma, minocycline reduced death rate and lesion area, but did not remarkably improve behavior and neuron loss.</p><p><b>CONCLUSION</b>Minocycline only has an incomplete neuroprotective effect on different brain traumatic injuries in rats and mice.</p>

Photomacrography of brain surface for evaluating blood-brain barrier disruption within 24 h after focal cerebral ischemia in mice / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To establish a new photomacrographic analysis of morphological changes on brain surface to evaluate blood-brain barrier (BBB) disruption.</p><p><b>METHODS</b>Permanent focal cerebral ischemia was induced by middle cerebral artery occlusion (MCAO) in mice. Brains were removed 10 min, 0.5, 1, 3, 6, 12 and 24 h after MCAO. The whole brains and brain slices were photographed by a digital camera. BBB disruption was evaluated by hemorrhage and traced Evans blue (EB) on the brain surface. Fluoremetric quantitation of EB and water content in the brains were also performed at various time points.</p><p><b>RESULT</b>Photomacrographic morphological analysis showed that hemorrhage and traced EB on the surface of the brains significantly increased from 3 h after focal cerebral ischemia,which were correlated to the results in the brain slices. EB content in the ischemic hemispheres was significantly increased from 0.5 h after MCAO, and water content was increased from 1 h after MCAO.</p><p><b>CONCLUSION</b>Photomacrographic measurement is a simple and useful method for evaluating BBB disruption semi-quantitatively, and can detect BBB disruption earlier after focal cerebral ischemia in mice.</p>

Macrophotographic measurement of brain surface area for evaluating brain edema after focal cerebral ischemia in mice / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To establish a new macrophotographic measurement of brain surface area to evaluate brain edema after focal cerebral ischemia in mice.</p><p><b>METHODS</b>Permanent focal cerebral ischemia was induced by middle cerebral artery occlusion (MCAO) in mice. The brains were removed 10,30 min,1,3,6,12 and 24 h after MCAO, and photographed in dorsal and lateral views by a digital camera. Then, 6 coronal slices of 1 mm thick were cut and photographed. Finally, the water content of brain tissue was measured by heating at 110 degrees C for 24 h. The left and right hemisphere areas of the brains and the brain slices were analyzed and calculated by MedBrain 2 imaging analyzer to evaluate brain edema.</p><p><b>RESULT</b>The macrophotographic measurement showed that the ischemic hemisphere areas significantly increased from 1 h after focal cerebral ischmia, which was similar to the measurement of water content. This measurement for brain edema correlated well with those of water content and brain slice volume.</p><p><b>CONCLUSION</b>The macrophotographic measurement is an objective and quantitative method for evaluating brain edema after focal cerebral ischemia.</p>