BACKGROUND: Currently, lung cancer is one of the malignant tumors with a high morbidity and mortality all over the world. However, the exact mechanisms underlying lung cancer progression remain unclear. The tumor necrosis factor receptor associated factor (TRAF) family members are cytoplasmic adaptor proteins, which function as both adaptor proteins and ubiquitin ligases to regulate diverse receptor signalings, leading to the activation of nuclear factor kappa-B (NF-κB), mitogen-activated protein kinase (MAPK) and interferon regulatory factor (IRF) signaling. The aim of this study was to investigate the expression of TRAFs in different tissues and cancer types, as well as its mRNA expression, protein expression, prognostic significance and functional enrichment analysis in non-small cell lung cancer (NSCLC), in order to provide new strategies for the diagnosis and treatment of NSCLC. METHODS: RNA sequencing data from the The Genotype-Tissue Expression database was used to analyze the expression patterns of TRAF family members in different human tissues. RNA sequencing data from the Cancer Cell Line Encyclopedia database was used to analyze the expression patterns of TRAF family members in different types of cancer cell lines. RNA sequencing data from the The Cancer Genome Atlas (TCGA) database was used to analyze the mRNA levels of TRAF family members across different types of human cancers. Immunohistochemistry (IHC) analyses from HPA database were used to analyze the TRAF protein levels in NSCLC [lung adenocarcinoma (LUAD) and lung squamous carcinoma (LUSC)]. Overall survival analysis was performed by Log-rank test using original data from Kaplan-Meier Plotter database to evaluate the correlation between TRAF expressions and prognosis. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses were performed on the TRAF family-related genes using RNA sequencing data from the TCGA database for NSCLC. The correlation between the expression levels of TRAF family members and the tumor immune microenvironment was analyzed using the ESTIMATE algorithm based on RNA sequencing data from the TCGA database. RESULTS: The TRAF family members exhibited significant tissue-specific expression heterogeneity. TRAF2, TRAF3, TRAF6 and TRAF7 were widely expressed in most tissues, while the expressions of TRAF1, TRAF4 and TRAF5 were restricted to specific tissues. The expressions of TRAF family members were highly specific among different types of cancer cell lines. In mRNA database of LUAD and LUSC, the expressions of TRAF2, TRAF4, TRAF5 and TRAF7 were significantly upregulated; while TRAF6 did the opposite; moveover, TRAF1 and TRAF3 only displayed a significant upregulation in LUAD and LUSC, respectively. Except for TRAF3, TRAF4 and TRAF7, other TRAF proteins displayed an obviously deeper IHC staining in LUAD and LUSC tissues compared with normal tissues. Additionally, patients with higher expression levels of TRAF2, TRAF4 and TRAF7 had shorter overall survival; while patients with higher expression levels of TRAF3, TRAF5 and TRAF6 had significantly longer overall survival; however, no significant difference had been observed between TRAF1 expression and the overall survival. TRAF family members differentially regulated multiple pathways, including NF-κB, immune response, cell adhesion and RNA splicing. The expression levels of TRAF family members were closely associated with immune cell infiltration and stromal cell content in the tumor immune microenvironment, with varying positive and negative correlations among different members. CONCLUSIONS TRAF family members exhibit highly specific expression differences across different tissues and cancer types. Most TRAF proteins exhibit upregulation at both mRNA and protein levels in NSCLC, whereas, only upregulated expressions of TRAF2, TRAF4 and TRAF7 predict worse prognosis. The TRAF family members regulate processes such as inflammation, immunity, adhesion and splicing, and influence the tumor immune microenvironment.
Humans ; Carcinoma, Non-Small-Cell Lung/pathology* ; Lung Neoplasms/mortality* ; Prognosis ; Gene Expression Regulation, Neoplastic ; Tumor Necrosis Factor Receptor-Associated Peptides and Proteins/metabolism*

Objective To investigate the value of methylation levels of Discs large-associated protein5(DLGAP5)and Ras association domain family 1A(RASSF1A)in peripheral blood in the differential diagnosis of benign and malignant lung nodules.Methods 110 patients with pulmonary nodules admitted to the Seventh People's Hospital of Hebei Province from December 2022 to June 2024 were selected as study subjects.Based on the diagnosis confirmed by pathological examination,52 patients with malignant lung nodules were included in the malignant group and 58 patients with benign lung nodules were included in the benign group.The methylation levels of DLGAP5 and RASSF1A genes in peripheral blood were determined by quantitative real-time polymerase chain reaction(qRT-PCR).The diagnostic value of methylation of DLGAP5 and RASSF1A genes in peripheral blood on the benign and malignant nature of lung nodules and their relationship with clinicopathological characteristics of patients with malignant lung nodules were analyzed.Spearman's method was used to analyze the correlation between DLGAP5 and RASSF1A gene methylation levels and clinicopathological features.Results Compared with the benign group,the methylation levels of DLGAP5 and RASSF1A genes were higher in the peripheral blood of patients in the malignant group,and the differences were statistically significant(χ2=27.010,24.350,all P<0.001).The sensitivities of DLGAP5 methylation and RASSF1A methylation alone in the detection of malignant lung nodules were 61.54%,67.31%,and the specificity were 86.21%,79.31%,and the accuracies were 74.55%,73.63%,respectively.The sensitivity(84.62%)and accuracy(83.64%)of the combined two-gene methylation test for malignant lung nodules were better than that of single gene methylation(Z=2.816,2.497,all P<0.05).The concordance Kappa values of DLGAP5 methylation and RASSF1A methylation alone and in combination with the pathological results were 0.583,0.569 and 0.712,respectively.The nodules size,histological differentiation degree,lymph nodes and spiculation sign of patients with malignant pulmonary nodules were correlated with DLGAP5 and RASSF1A methylation levels,and the differences were statistically significant(χ2DLGAP5=4.644～14.981,χ2RASSF1A=4.293～12.629,all P<0.05).Correlation analysis showed that the methylation levels of DLGAP5 and RASSF1A genes were positively correlated with nodule size,histologic differentiation,lymph node metastasis and spiculation sign(rDLGAP5=0.512～0.683,rRASSF1A=0.527～0.691,all P<0.05).Conclusion The methylation levels of DLGAP5 and RASSF1A gene in peripheral blood were closely related to histological differentiation,lymph node metastasis and clinical staging.The combined detection of the two has high diagnostic value fro the benign and malignant nature of pulmonary nodules.

Objective To investigate the value of methylation levels of Discs large-associated protein5(DLGAP5)and Ras association domain family 1A(RASSF1A)in peripheral blood in the differential diagnosis of benign and malignant lung nodules.Methods 110 patients with pulmonary nodules admitted to the Seventh People's Hospital of Hebei Province from December 2022 to June 2024 were selected as study subjects.Based on the diagnosis confirmed by pathological examination,52 patients with malignant lung nodules were included in the malignant group and 58 patients with benign lung nodules were included in the benign group.The methylation levels of DLGAP5 and RASSF1A genes in peripheral blood were determined by quantitative real-time polymerase chain reaction(qRT-PCR).The diagnostic value of methylation of DLGAP5 and RASSF1A genes in peripheral blood on the benign and malignant nature of lung nodules and their relationship with clinicopathological characteristics of patients with malignant lung nodules were analyzed.Spearman's method was used to analyze the correlation between DLGAP5 and RASSF1A gene methylation levels and clinicopathological features.Results Compared with the benign group,the methylation levels of DLGAP5 and RASSF1A genes were higher in the peripheral blood of patients in the malignant group,and the differences were statistically significant(χ2=27.010,24.350,all P<0.001).The sensitivities of DLGAP5 methylation and RASSF1A methylation alone in the detection of malignant lung nodules were 61.54%,67.31%,and the specificity were 86.21%,79.31%,and the accuracies were 74.55%,73.63%,respectively.The sensitivity(84.62%)and accuracy(83.64%)of the combined two-gene methylation test for malignant lung nodules were better than that of single gene methylation(Z=2.816,2.497,all P<0.05).The concordance Kappa values of DLGAP5 methylation and RASSF1A methylation alone and in combination with the pathological results were 0.583,0.569 and 0.712,respectively.The nodules size,histological differentiation degree,lymph nodes and spiculation sign of patients with malignant pulmonary nodules were correlated with DLGAP5 and RASSF1A methylation levels,and the differences were statistically significant(χ2DLGAP5=4.644～14.981,χ2RASSF1A=4.293～12.629,all P<0.05).Correlation analysis showed that the methylation levels of DLGAP5 and RASSF1A genes were positively correlated with nodule size,histologic differentiation,lymph node metastasis and spiculation sign(rDLGAP5=0.512～0.683,rRASSF1A=0.527～0.691,all P<0.05).Conclusion The methylation levels of DLGAP5 and RASSF1A gene in peripheral blood were closely related to histological differentiation,lymph node metastasis and clinical staging.The combined detection of the two has high diagnostic value fro the benign and malignant nature of pulmonary nodules.

Objective:To investigate the clinical phenotypes, muscle magnetic resonance imaging (MRI) and pathological changes, and genetic characteristics of myfibrillar myopathies (MFMs) cuased by MYOT gene mutation. Methods:(1) The clinical data of a MFMs family caused by a de novo frameshift mutation in MYOT gene admitted to Department of Neurology, Jiaozuo People's Hospital Affiliated to Xinxiang Medical University in February 2021 were collected. Electromyography, muscle MRI, and pathological examination were used to confirm the changes of the muscle lesions. MYOT gene mutation in the proband and other patients was detected by next generation sequencing (NGS) and Sanger sequencing, respectively. The 3D structure models of myotilin protein before and after gene mutation were predicted by AlphaFold3 and pymol3. (2) Literature on MFMs caused by MYOT gene mutation was searched from Pubmed and China National Knowledge Infrastructure from the establishment of these databases to July 2024; clinical and genetic characteristics of MFMs caused by MYOT gene mutation were summarized. Results:(1) In the 9 patients from this family, 8 had onset in adolescence (16-20 years old). Unilateral or bilateral hand muscle weakness as the first symptoms appeared in most patients, and then, hand muscle atrophy gradually appeared and slowly progressed to the proximal limbs. Electromyography showed myogenic damage. Muscle MRI showed patchy long T1 and long T2 signal intensity in the bilateral anterior tibial muscles. Muscle pathological staining showed typical rimbed vacuoles, cytoplasm, smear-like muscle fibers and desmin abnormal deposition in some muscle fibers; electron microscopy revealed disorganized myofibril structures, focal myofibril lysis, Z-band streaming, and subsarcolemmal or myofibril mitochondrial accumulation. Heterozygous mutation in MYOT gene c.680_683del (p.Val227GlufsTer10) locus was noted in 8 patients and daughter of the proband. Bioinformatics analysis suggested that MYOT gene c.680_683del mutation could cause premature termination of myotilin translation, leading to the production of a truncated protein, thereby disrupting its normal structure and function. (2) Eighty-nine patients with MFMs caused by MYOT gene mutation in previous literature mainly manifested as chronic progressive weakness of the distal or proximal limbs, with some involving the myocardium, respiratory muscles, or peripheral nerves. A total of 12 MYOT gene mutations were identified, with p. Ser60phe being the most common mutation. Except for p.Tyr4_his9del, being an in-frame mutation, the others were missense mutations. Conclusion:MFMs caused by MYOT gene mutation exhibit obvious clinical heterogeneity, characterized by very slow progression of muscle weakness; MYOT gene locus c.680_683del (p.Val227GlufsTer10) is a de novo heterozygous mutation.

Objective:To investigate the clinical phenotypes, muscle magnetic resonance imaging (MRI) and pathological changes, and genetic characteristics of myfibrillar myopathies (MFMs) cuased by MYOT gene mutation. Methods:(1) The clinical data of a MFMs family caused by a de novo frameshift mutation in MYOT gene admitted to Department of Neurology, Jiaozuo People's Hospital Affiliated to Xinxiang Medical University in February 2021 were collected. Electromyography, muscle MRI, and pathological examination were used to confirm the changes of the muscle lesions. MYOT gene mutation in the proband and other patients was detected by next generation sequencing (NGS) and Sanger sequencing, respectively. The 3D structure models of myotilin protein before and after gene mutation were predicted by AlphaFold3 and pymol3. (2) Literature on MFMs caused by MYOT gene mutation was searched from Pubmed and China National Knowledge Infrastructure from the establishment of these databases to July 2024; clinical and genetic characteristics of MFMs caused by MYOT gene mutation were summarized. Results:(1) In the 9 patients from this family, 8 had onset in adolescence (16-20 years old). Unilateral or bilateral hand muscle weakness as the first symptoms appeared in most patients, and then, hand muscle atrophy gradually appeared and slowly progressed to the proximal limbs. Electromyography showed myogenic damage. Muscle MRI showed patchy long T1 and long T2 signal intensity in the bilateral anterior tibial muscles. Muscle pathological staining showed typical rimbed vacuoles, cytoplasm, smear-like muscle fibers and desmin abnormal deposition in some muscle fibers; electron microscopy revealed disorganized myofibril structures, focal myofibril lysis, Z-band streaming, and subsarcolemmal or myofibril mitochondrial accumulation. Heterozygous mutation in MYOT gene c.680_683del (p.Val227GlufsTer10) locus was noted in 8 patients and daughter of the proband. Bioinformatics analysis suggested that MYOT gene c.680_683del mutation could cause premature termination of myotilin translation, leading to the production of a truncated protein, thereby disrupting its normal structure and function. (2) Eighty-nine patients with MFMs caused by MYOT gene mutation in previous literature mainly manifested as chronic progressive weakness of the distal or proximal limbs, with some involving the myocardium, respiratory muscles, or peripheral nerves. A total of 12 MYOT gene mutations were identified, with p. Ser60phe being the most common mutation. Except for p.Tyr4_his9del, being an in-frame mutation, the others were missense mutations. Conclusion:MFMs caused by MYOT gene mutation exhibit obvious clinical heterogeneity, characterized by very slow progression of muscle weakness; MYOT gene locus c.680_683del (p.Val227GlufsTer10) is a de novo heterozygous mutation.

Infant, Newborn ; Humans ; Retrospective Studies ; Case-Control Studies ; Tracheotomy ; Respiration, Artificial

Objective To investigate the clinical,brain MRI and MT-ATP6 gene variation characteristics of late-onset Leigh syndrome (LS). Methods The clinical data,brain imaging and genetic test results of two patients with late-onset Leigh syndrome in a family were collected in detail,and discussed in combination with the literature. Results The proband,male,9 years old,complained of "bilateral ptosis,unstable walking for half a year,abnormal mental behavior for 4 months".He was admitted to our hospital on April 10,2020.Patient was born normally,his physique and intelligence was worse that of his peers since childhood.Six months ago,the patient had bilateral ptosis and double vision,and he gradually developed unstable walking and difficulty in running.Brain MRI examination showed abnormal signals in bilateral basal ganglia and brain stem.4 months ago,he had intermittent nausea and vomiting,abnormal mental behavior without any inducement.Two months ago,he suddenly fell into a coma and twitched after catching a cold and fever.He was rescued and treated in a local hospital.His vital signs were stable and transferred to our hospital.Physical examination:short stature,thin physique,coma with eyes open.Bilateral ptosis covers more than half of the pupil,bilateral eyeball center fixation,dysphagia,nasal feeding diet.Muscle tension in the limbs was high,tendon reflex was active,and the limbs were flexing,Bilateral pathological signs were positive.Family history:the child's parents are not consanguineous marriage,the father is in good health,the mother is short and has a history of mental and behavioral abnormalities.The proband's brother,14 years old,was less developed than his peers.At the age of 7,he developed ptosis,unstable walking and no history of coma and convulsions.No ragged red fibers were found in skeletal muscle pathological biopsies of the proband and his brother.The mutation of MT-ATP6 gene 9176T>C was found by mitochondrial genome detection. Conclusions The clinical phenotype of late-onset Leigh syndrome is heterogeneous,which may be accompanied by mental and behavioral abnormalities and cognitive impairment.Brain MRI is characterized by abnormal signals in bilateral basal ganglia and midbrain periaqueductal region.Gene detection is an important basis for the diagnosis of Leigh syndrome.

Objective:To summarize the characteristics of clinical, muscle pathology and gene mutation of late-onset reducing body myopathy caused by FHL1 gene mutation, in order to improve clinicians′ understanding of this disorder. Methods:The clinical, muscle pathology and muscle magnetic resonance imaging data of the proband from a family diagnosed as reducing body myopathy in Jiaozuo People′s Hospital in December 2021 were collected. Genetic tests and pedigree verification were conducted on the proband and her son.Results:The proband was a 59-year-old female with progressive, asymmetrical limb weakness and muscular atrophy. Her mother, sister and brother had similar symptoms. Electromyography showed myogenic and neurogenic damage. Muscle magnetic resonance imaging indicated that the lesion mainly involved the posterior muscles of the thigh and calf, as well as the gluteus maximus. The muscle pathology showed eosinophilic granular inclusion bodies and rimmed vacuoles in the muscle fibers of the lesion. The structure of myofibrils was disordered and abnormal protein deposition was observed. The gene sequencing showed the FHL1 gene p.C150S heterozygous variation. Conclusions:Late-onset reducing body myopathy is characterized by progressive asymmetric proximal limb muscle weakness, partially involving distal limb muscles and gluteus maximus. Muscle pathology shows the characteristic pathological changes of many kinds of myofibrillar myopathies. FHL1 gene mutation is an important basis for diagnosis.

Objective:To study the outcomes and post-discharge follow-up of neonatal tracheotomy in neonatal intensive care unit(NICU).Methods:This study included patients who were admitted to NICU in Beijing Children′s Hospital from January, 2016 to August, 2021, and less than 28 days or 44 weeks(corrected age)on admission, and required tracheotomy.The patients were divided into tracheotomy group and the non-tracheotomy group (the parents signed to refuse the tracheotomy) according to whether perform tracheotomy.Demographic data, general hospitalization information, diagnosis, indications for tracheotomy, follow-up outcomes at 3/6/12 months of age after discharge of patients were collected and analyzed.Results:Totally 26 patients were included in this study, 14 cases in tracheotomy group and 12 cases in non-tracheotomy group.The average gestational age was(37.7±3.80)weeks and(38.99±1.83)weeks, and birth weight was(2 823.57±948.89)g and (3 320.83±378.76)g, respectively.There were no significant differences in sex, gestational age, birth weight, age on admission, weight on admission, age at diagnosis, ratio of endotracheal intubation for respiratory support on admission between two groups( P>0.05). The commonest indications of tracheotomy group were bilateral vocal cord paralysis(50.0%) and congenital anomaly/defect of throat/larynx(21.4%), and the commonest indications of non-tracheotomy group were bilateral vocal cord paralysis(50.0%) and vocal cord/subglottic mass(25.0%), and there was no significant difference between two groups( P>0.05). The rate of discharge-against-medical order of tracheotomy and non-tracheotomy group was 7.14% and 66.67%( P=0.003), respectively.The total follow-up rate of tracheotomy and non-tracheotomy group was 88.9% and 38.9%, while the follow-up rates at 3 months, 6 months, and 12 months were 100.0% vs. 50.0%, 83.3% vs. 41.7%, and 81.8% vs. 25.0%, respectively, whose differences were statistically significant(all P<0.05). In the 14 cases of tracheotomy group, 3 cases died, 4 cases successfully removed the tracheal cannula, 5 cases did not remove the tracheal cannula, and 2 cases were lost. Conclusion:Bilateral vocal cord paralysis is the commonest indication of neonatal tracheotomy.Parents′ compliance in the tracheotomy group is significantly higher than that in non-tracheotomy group.To give caring skill training for parents of neonates with tracheotomy before discharge is beneficial for improving the overall prognosis of children.

Objective:To investigate the clinical characteristics, skeletal muscle pathologies and gene variations of chronic progressive external ophthalmoplegia (CPEO).Methods:Sixteen patients with conformed CPEO, admitted to our hospital from January 1997 to December 2021, were chosen. Their clinical data such as onset age and course of diseases and muscle pathological examination results were collected and their gene variation characteristics were analyzed.Results:The initial symptom in all 16 patients was ptosis of varying degrees; 15 patients were with eye movement disorder, 6 with diplopia, 4 with proximal limb weakness, and 3 with dysphagia and dysarthria. Among the 16 patients, electromyography showed myogenic damage in 7 patients, myogenic combined with neurogenic damage in 1 patient, neurogenic damage in 1 patient, and normal in 7 patients. Skeletal muscle biopsies indicated that 14 patients were with ragged red fibers (RRF), 11 patients had cytochrome C oxidase (COX)-negative muscle fibers, 3 patients had a small amount of degenerated and necrotic myofibers with mononuclear phagocytic infiltration. Immunohistochemical staining indicated infiltration of CD8 and CD68 positive lymphocytes. Ten patients accepted genetic test, indicating 6 patients with single large fragment deletion of mitochondrial DNA (mtDNA), 1 patient with mtDNA point mutation, 1 patient with nucleosomal DNA (nDNA) point mutation, and 2 patients without pathogenicity variation clearly associated with clinical phenotype. Electron microscopy in 5 patients showed that abnormal mitochondrial aggregation was noted in 4 patients under the sarcolemma and among the myofibrils.Conclusion:In addition to ptosis and eye movement disorders, a small number of patients with CPEO may be accompanied by dysphagia and limb weakness; and single large fragment deletion of mtDNA is the main mutation form of CPEO.