Generalized machine learning based on multi-omics data to profile the effect of ferroptosis pathway on prognosis and immunotherapy response in patients with bladder cancer.

INTRODUCTION: Bladder cancer (BLCA) affects millions of people worldwide, with high rates of incidence and mortality. Ferroptosis proves to be a novel form of cell death process that is triggered by oxidative stress. METHODS: We procured a total of 25 single nuclear RNA-seq (snRNA-seq) samples from GSE169379 in GEO database. We obtained different cohorts of BLCA patients from the TCGA and GEO databases for model training and validation. A total of 369 ferroptosis-related genes (FRGs) were selected from the FerrDb database. AUCell analysis was performed to assign ferroptosis scores to all the cell types. Weighted Gene Co-Expression Network Analysis (WGCNA), COX, and LASSO regression analysis were conducted to retain and finalize the genes of prognostic values. Various bioinformatic approaches were utilized to depict immune infiltration profile. We conducted a series of colony formation analysis, flow cytometry and western blot (WB) analysis to determine the role of SKAP1 in BLCA. RESULTS: We divided the cells into high ferroptosis group and low ferroptosis group according to ferroptosis activity score, and then screened 2150 genes most associated with ferroptosis by differential expression analysis, which are related to UV-induced DNA damage, male hormone response, fatty acid metabolism and hypoxia. Subsequently, WGCNA algorithm further screened 741 ferroptosis related genes from the 2150 genes for the construction of prognostic model. Lasso-Cox regression analysis was used to construct the prognostic model, and the prognostic model consisting of 6 genes was obtained, namely JUN, SYT1, MAP3K8, GALNT14, TCIRG1, and SKAP1. Next, we constructed a nomogram model that integrated clinical factors to improving the accuracy. In addition, we performed drug sensitivity analyses in different subgroups and found that Staurosporine, Rapamycin, Gemcitabine, and BI-2536 may be candidates for the drugs treatment in high-risk populations. The ESTIMATE results showed higher stromal scores, immune scores, and ESTIMATE scores in the low-risk group, indicating a higher overall immunity level and immunogenicity of tumor microenvironment (TME) in this group, and tumor immune dysfunction and exclusion (TIDE) analysis confirmed a better response to immunotherapy in the low-risk group. Finally, we selected the oncogene SKAP1 in the prognostic gene for in vitro validation, and found that SKAP1 directly regulated BLCA cell proliferation and apoptosis. CONCLUSION: We identified a set of six genes, JUN, SYT1, MAP3K8, GALNT14, TCIRG1, and SKAP1, that exhibited significant potential in stratification of BLCA patients with varying prognosis. In addition, we uncovered the direct regulatory effect of SKAP1 on BLCA cell proliferation and apoptosis, shedding some light on the role of FRGs in pathogenesis of BLCA.

TRIM33 protects osteoblasts from oxidative stress-induced apoptosis in osteoporosis by inhibiting FOXO3a ubiquitylation and degradation.

This study aimed to probe into the effect of TRIM33 on oxidative stress-induced apoptosis of osteoblasts in osteoporosis and to probe into the underlying mechanism. The apoptosis of osteoblasts was induced by H2 O2 treatment and tested by flow cytometry. A mouse osteoporosis model was conducted by ovariectomy (OVX). The function of TRIM33 was assessed by in vitro and in vivo experiments. The mechanism of TRIM33 was determined by immunoprecipitation, immunofluorescent staining and co-transfection experiments. Here, we found that TRIM33 expression was lessened in the osteoblasts of patients with osteoporosis and was positively correlated with the bone mineral density of these patients. FOXO3a and TRIM33 were co-localized in the osteoblasts nuclei. TRIM33 silence boosted FOXO3a degradation in normal osteoblasts, while TRIM33 overexpression restrained FOXO3a degradation in H2 O2 -treated osteoblasts. The binding of TRIM33 to CBP and its overexpression restrained CBP-mediated FOXO3a acetylation, thereby attenuating FOXO3a ubiquitylation. The H2 O2 -induced apoptosis of osteoblasts was restrained by TRIM33 overexpression, while the FOXO3a knockdown reversed this trend. The in vivo experiments corroborated that TRIM33 overexpression attenuated the OVX-driven impacts in mice. In general, our findings expounded that TRIM33 protected osteoblasts against oxidative stress-induced apoptosis in osteoporosis and that the underlying mechanism was the restraint of FOXO3a ubiquitylation and degradation.

Clinicopathological characteristics and survival of malignant fibrous histiocytoma of the bone: A population-based study using the SEER database.

Malignant fibrous histiocytoma of the bone (MFH-B) is an extremely rare and aggressive malignancy. The clinicopathological characteristics and prognosis of patients with MFH-B have not been defined. We conducted a retrospective study using the data of all MFH-B patients from the Surveillance, Epidemiology and End Results (SEER) database between 1975 and 2016. Initially, the clinicopathological characteristics were described. The difference in prognosis between patients with MFH-B and those with osteosarcoma was compared using propensity score matching analysis. Then, the features affecting the prognosis of patients with MFH-B were further determined using Cox regression analysis. A total of 318 patients with MFH-B were identified. The median overall survival (mOS) of all 318 patients with MFH-B was 29.0 months. The 1-, 3-, 5-, and 10- year survival rates were 67.4%, 53.6%, 38.7%, and 28.7%, respectively. The multivariate Cox regression analysis showed that older age, distant metastases, and flat bone lesion were independent factors for worse prognosis, whereas surgery was an independent factor for favorable survival, and this intervention could decrease risk of death by 61% (HR = 0.39, 95% CI 0.28-0.54). Apart from this, the prognosis of patients with MFH-B was significantly worse than that of patients with osteosarcoma in both unmatched and matched cohorts. In conclusion, MFH-B is a rare malignant bone cancer, with relatively worse prognosis than osteosarcoma. Older age, distant metastases, flat bone lesion, and surgery were independently associated with prognosis. In order to understand this disease more thoroughly and accurately, more cases with adequate information are required in the future.

The association of XRCC1 polymorphism with osteosarcoma risk, clinicopathologic features, and prognosis in a Chinese Han population.

INTRODUCTION: The association of single-nucleotide polymorphisms at X-ray repair cross-complementing group-1 (XRCC1) with osteosarcoma (OS) development has not been fully clear to date. The aim of the present study is to evaluate the association of XRCC1 polymorphisms with risk, clinicopathologic features, and prognosis in Chinese OS patients. METHODS: A total of 146 patients with primary OS and 248 age- and gender-matched controls were included in the present study. The frequencies of four XRCC1 polymorphisms (rs25487, rs1799782, rs25489, and rs3213245) were determined between OS patients and controls. The association of XRCC1 polymorphism with clinicopathologic characteristics, prognosis, and XRCC1 expression was further evaluated. RESULTS: Compared with TT genotype, individuals carrying the minor C allele (TC+ CC) of rs3213245 had significantly increased risk of OS development (OR =1.83, 95% CI 1.14-3.00). OS patients carrying TC genotype and C allele at rs3213245 were more likely to be with larger tumor size and metastasis. Survival analysis demonstrated that OS patients carrying C allele (TC + CC) at rs3213245 had shorter survival time than those with TT genotype. The T to C substitution at rs3213245 could decrease XRCC1 gene transcriptional activity in vitro. XRCC1 mRNA and protein expression levels were lower in OS patients carrying TC or CC genotype at rs3213245. Besides, no significant association of rs25487, rs1799782, and rs25489 with OS was observed. CONCLUSION: In conclusion, these findings revealed that XRCC1 rs3213245 polymorphism was associated with increased risk of OS, which could affect XRCC1 expression in vitro and in vivo.

Th17 and IL-17 exhibit higher levels in osteonecrosis of the femoral head and have a positive correlation with severity of pain.

OBJECTIVE: Synovitis associated with osteonecrosis of the femoral head (ONFH) is responsible for several clinical symptoms. However, the mechanisms underlying synovitis and the inflammatory environment remain unclear. This study analyzed the proinflammatory mediation expression of IL-17 and Th17, which perform key functions in regulating inflammatory processes in the inflamed synovium and peripheral blood in ONFH. METHODS: Synovial fluid from the hips of 23 patients and 5 controls was collected during surgery, and peripheral blood samples were obtained from 34 patients and 9 controls. The expression of IL-17 in the synovium was detected by immunohistochemistry, and the levels of Th17 and IL-17 in the blood were measured by flow cytometry and ELISA. Pain assessment was performed for all the patients and controls. RESULTS: An inflamed synovium was characterized by increased leukocyte infiltration and IL-17 expression in comparison with the control. Preoperative levels of Th17 and IL-17 were significantly higher in the peripheral blood of the ONFH group than those in the controls. The symptoms were also positively correlated with the Th17 levels of the ONFH patients. CONCLUSION: Th17 cells were recruited to an inflamed synovium, and inflammatory cytokine IL-17 was expressed at an increased level in the hip synovium of ONFH patients, which possibly contributed to clinical syndrome development. Overall, this study will help in identifying new therapeutic strategies for ONFH, especially the targeting of IL-17 to decrease inflammation and pain. < p > < /p >.

rs1760944 Polymorphism in the APE1 Region is Associated with Risk and Prognosis of Osteosarcoma in the Chinese Han Population.

The effects of single nucleotide polymorphisms (SNPs) at APE1 have been investigated in several types of cancer. However, no reports of the association of APE1 polymorphisms with osteosarcoma (OS) have been published. The present study was designed to determine whether APE1 polymorphisms (rs1130409, rs1760944, rs1760941, rs2275008, rs17111750) are associated with OS. A 2-stage case-control study was performed in a total of 378 OS patients and 616 normal controls. Individuals carrying TG and GG genotypes had significantly lower risk of developing OS than those with the WT genotype TT at rs1760944 (OR = 0.65, 95%CI 0.49-0.86; OR = 0.50, 95%CI 0.34-0.74, respectively). OS patients with allele G at rs1760944 were less susceptible to low differentiation tumor and metastasis (OR = 0.73, 95%CI 0.54-0.98; OR = 0.63, 95%CI 0.43-0.92, respectively). Kaplan-Meier curves and log-rank results revealed that OS patients harboring genotype GG and G allele at rs1760944 had better survival (P < 0.001 for both). In addition, the APE1 protein was underexpressed in individuals who carried G allele at rs1760944. This study suggested that APE1 rs1760944 polymorphism is associated with decreased risk of developing OS and better survival of OS patients.

MiR-125a Rs12976445 Polymorphism is Associated with the Apoptosis Status of Nucleus Pulposus Cells and the Risk of Intervertebral Disc Degeneration.

BACKGROUND: Spinal degenerative diseases are a major health problem and social burden worldwide. Intervertebral disc degeneration (IDD) is the pathological basis of spinal degenerative diseases and is characterized by loss of nucleus pulposus cells due to excessive apoptosis caused by various factors. MicroRNAs (miRNAs) have been reported to be functionally involved in the control of apoptosis. METHODS: computational analysis and luciferase assay were used to identify the target of miR-125a, and cell culture, transfection were used to confirm such relationship. Sequencing was used to determine the genotype of each participant. RESULTS: We confirmed the previous report that the presence of the minor allele (T) of rs12976445 polymorphism significantly downregulated the expression level of miR-125a in nucleus pulposus cells, leading to less efficient inhibition of its target gene. We also validated TP53INP1 as a target of miR-125a in nucleus pulposus cells using a dual luciferase reporter system, and the transfection of miR-125a significantly reduced the expression of TP53INP1. The expression level of TP53INP1 was significantly lower in nucleus pulposus cells genotyped as CT or TT than in those genotyped as CC, and the apoptosis rate was consistently lower in the CC group than in the nucleus pulposus cells collected from individuals carrying at least one minor allele of rs12976445 polymorphism. To study the association between rs12976445 polymorphism and the risk of IDD, we enrolled 242 patients diagnosed with IDD and 278 normal controls, and significant differences were noted regarding the genotype distribution of rs12976445 between the IDD and the control groups (OR = 2.69, 95% C.I. = 1.88-3.83, p < 0.0001). In summary, rs12976445 polymorphism is significantly associated with the risk of IDD in the Chinese population. CONCLUSION: The present study indicated that miR-125a is a promising potential target for patients with IDD in clinical practice.

Therapeutic effects of PKP on chronic painful osteoporotic vertebral compression fractures with or without intravertebral cleft.

This study aims to investigate clinical effects of protruding after percutaneous vertebral plasty (PKP holds) for treatment of chronic painful osteoporotic vertebral compression fractures (CPOVCFs), and assess its safety and effectiveness. One hundred and eighteen CPOVCFs patients were included, and analyzed and followed up therapeutic effects of PKP holds. All of these patients were divided into NIVC (73 cases) and IVC (45 cases) group. Comparing operation time, bone cement perfusion, pain intensity modulus visual grading scoring, Oswestry disability index, fracture fanterior relative height, local convex Angle, and surgical complications between the two groups. There were no significant differences between two groups for operation time of PKP holds (P>0.05). Amount of bone cement injection in IVC groups was significantly higher compared to NIVC group (P<0.05). Pain intensity visual modulus ratings and Oswestry disability index were significantly improved in both groups after surgery and postoperative follow-up period compared to pre-operation (P<0.05). Fracture fanterior margin relative height and local convex angle were also significantly improved (P<0.05). Fracture fanterior margin relative height was also significantly improved for IVC compared to NIVC group (P<0.05). There were no differences for bone cement leakage rate and adjacent segment fracture between two groups (P>0.05). But there was a case of bone cement loose, which needs to be treated for further step. In conclusion, PKP holds is a kind of effective and safe method for the IVC and NIVC patients. Especially for the IVC patients, PKP holds can significantly improve the fracture fanterior margin relative height.

Prevention and treatment of bone cement-related complications in patients receiving percutaneous kyphoplasty.

OBJECTIVE: This study is to investigate the complications and preventive measures of bone cement in percutaneous kyphoplasty for the treatment of osteoporotic vertebral compression fractures. METHODS: A retrospective analysis of 134 cases of osteoporotic vertebral compression fracture patients treated with percutaneous kyphoplasty, which involved 182 vertebral fractures, was performed. The bone cement-related complications, the reasons causing these complications, and the preventive measures were investigated. RESULTS: Successful operation was performed in all patients. Operative bone cement toxic reaction occurred in 1 patient. Different degrees of leakage of bone cement were found in 21 patients with 25 fractures and 1 case needed operation treatment. During the long term follow-up, bone necrosis absorption and cement drifted away which needed operation treatment was found in 1 patient with vertebral fracture. Thirteen adjacent vertebral fractures were found in 11 patients and 10 patients underwent operation treatment again. CONCLUSIONS: With careful surgery, complications associated with bone cement could be greatly reduced.

Three-dimensional image navigation system-assisted anterior cervical screw fixation for treatment of acute odontoid fracture.

This study is to investigate the role of three-dimensional image navigation system for surgical treatment of odontoid fracture. A total of 21 patients were enrolled in this study. The anterior cervical hollow screw fixation was performed for treatment of acute odontoid fracture under monitoring of isocentric C-arm three-dimensional navigation system (Iso-C 3D) navigation system. The postoperative follow-up investigation duration was 13.8 ± 4.4 months. Twenty patients with odontoid fracture had bone union without intraoperative and postoperative complications. No loosening, dislocation or fracture of screw occurred. The average healing time was from 3 to 4 months. The cervical postoperative organ function evaluation of patients was scored by Smiley-Webster scoring methods. Eighteen cases were scored as excellent; 2 cases were scored as fine; 1 case was scored as good; and no case was scored as poor. Iso-C 3D image navigation system-assisted anterior cervical screw fixation is an effective approach for treatment of odontoid fracture.

Cervical curvature, spinal cord MRIT2 signal, and occupying ratio impact surgical approach selection in patients with ossification of the posterior longitudinal ligament.

OBJECTIVE: Factors impacting surgical options and outcomes in patients with cervical ossification of the posterior longitudinal ligament (OPLL) were explored. METHODS: A retrospective analysis was conducted of 127 eligible cervical OPLL patients (61 males, 66 females) aged 41-70 years (mean 55.2 years) selected from 152 total OPLL patients treated from 2002 to 2006, with 5-10-year (mean 6.8 years) follow-up. Patients underwent anterior subtotal corpectomy with ossification ligament resection (anterior surgery, n = 68) or posterior cervical double-door laminoplasty (posterior surgery, n = 59). Radiographic assessments of cervical curvature, T2-weighted MRI (MRIT2) signal, and OPLL occupying ratio were correlated with surgical strategy before surgery and at 1, 5 weeks, and 5 years. RESULTS: Lordosis increased following anterior surgery, though kyphosis improved by 10.3 %. The canal stenosis occupying ratio was >50 %, and short-term improvement following anterior surgery was significantly higher than posterior surgery (P > 0.0001). Superior neurological function was observed in patients with unchanged versus high spinal MRIT2 signals (P = 0.0434). No significant differences were observed in short-term outcomes between anterior and posterior surgeries in high spinal MRIT2 signal patients, but anterior surgery produced significantly better long-term outcomes at 1 week (P = 0.7564) and 1 year (P = 0.0071). Complications occurred in five anterior and three posterior surgeries. CONCLUSION: Preoperative assessment of cervical curvature, MRIT2 signal, and occupying ratio can be used to guide clinical surgical approach selection to potentially produce better long-term outcomes in patients with OPLL.