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*

ObjectiveTo explore the mechanism of Gancao Fuzitang （GCFZ）in inhibiting the bone destruction of collagen-induced arthritis （CIA） model in mice. MethodThirty male DBa/1J mice were randomly divided into normal group， CIA group， low-dose GCFZ group （GCFZ-L， 2.4 g·kg^-1）， high-dose GCFZ group （GCFZ-H， 4.8 g·kg^-1）， and methotrexate group （MTX， 1 mg·kg^-1）， with six mice in each group. The CIA model was induced by secondary immunization method. The arthritis index of mice in each group was observed and recorded， and the histopathological changes in ankle joint were observed by hematoxylin-eosin （HE） staining. The damage to ankle cartilage was detected by safranin O-fast green staining. Micro-CT scanning was used to detect the bone destruction of ankle joint， and the expression of nuclear factor-κB p65 （NF-κB p65）， p-NF-κB p65， inhibitory-κB kinase α/β （IKKα/β）， and p-IKKα/β was observed by immunohistochemical staining. ResultCompared with the normal group， the CIA group showed manifest joint swelling and increased arthritis index score （P<0.01）. Compared with the CIA group， the groups with drug intervention could inhibit joint swelling and reduce arthritis index score （P<0.05， P<0.01）. As revealed by HE staining and safranine O-green staining， compared with the CIA group， the groups with drug intervention could inhibit synovial invasion and reduce the destruction of articular cartilage. Micro-CT scanning analysis showed that compared with the CIA group， the GCFZ-H group and the MTX group showed reduced bone destruction scores （P<0.01）. The immunohistochemical results showed that compared with the normal group， the CIA group showed increased optical density values of NF-κB p65， p-NF-κB p65， IKKα/β， and p-IKKα/β（P<0.01）. Compared with the CIA group， the GCFZ-H group and the MTX group showed reduced optical density values of NF-κB p65， p-NF-κB p65， IKKα/β， and p-IKKα/β（P<0.05，P<0.01）. In the GCFZ-L group， only the NF-κB p65 optical density value decreased（P<0.01）. ConclusionGCFZ may inhibit bone destruction in CIA mice by regulating the NF-κB signaling pathway.

Objective:To study the expression levels of peripheral blood helper T cell 17 (Th17) cells and interleukin (IL)-17A in newly diagnosed acute myeloid leukemia (AML) patients and its significance.Methods:A total of 32 newly diagnosed AML patients in Binzhou People's Hospital of Shandong Province from September 2017 to January 2019 were treated as the study group, and 28 iron deficiency anemia patients were used as the control group. Flow cytometry (FCM) was used to detect the proportion of Th17 (CD3 + CD4 + IL-17 +) in CD4 + T cells (Th17 ratio) and the concentration of IL-17A in peripheral bloods for both groups. And then the correlation and significance of Th17 ratio and the concentration of IL-17A with proportion of bone marrow blast cells, chromosome karyotype in AML patients was also analyzed. Results:The proportion of Th17 cells in peripheral bloods of newly diagnosed AML patients was higher than that in the control group [(2.74±0.85)% vs. (1.02±0.12)%, t = 10.397, P < 0.01]; the concentration of IL-17A in the serums of AML patients was higher than that of the control group [(3.16±1.54) pg/ml vs. (2.22±0.21) pg/ml, t = 3.206, P = 0.002]. Th17 ratio and the concentration of IL-17A in patients with bone marrow blast cells percentage≥0.50 were higher than those in patients with bone marrow blast cells percentage <0.50. Th17 ratio and the concentration of IL-17A in the peripheral bloods for AML patients in the high-risk group was higher than that in the low-risk group, and the difference was statistically significant (both P < 0.05). There was no statistical difference between low-risk group and intermediate-group (both P > 0.05). Conclusions:The levels of Th17 cells and IL-17A in the peripheral bloods are associated with the proportion of bone marrow blast cells and cytogenetics/molecular genetics risk degree in AML patients. The regular detection of Th17 and IL-17A may help to monitor immune status, evaluate prognosis and provide the basis for immunotherapy of AML patients.

Objective:To describe the trends of loss of muscle fiber thickness of the arm and calf.Methods:From April 2019 to July 2019, 58 patients admitted to ICU of Qinghai Provincial People's Hospital were enrolled. During ICU stay, the muscle fiber thickness in bilateral upper arm (biceps brachii) and bilateral calf (gastrocephus muscle) were measured by bedside color ultrasound every day.Results:The muscle fiber thickness of biceps and gastrocnemius on the left and right were (1.52 ± 0.37), (1.50 ± 0.34), (1.53±0.39), (1.51 ± 0.37) mm, respectively. The thickness of muscle fibers were (1.45 ± 0.35), (1.46 ± 0.37), (1.44±0.33), (1.41 ± 0.32) mm, which were significantly decreased in 48 h admitted to ICU ( t values were 2.106-4.711, P<0.05 or 0.01); and (1.43 ± 0.36), (1.44 ± 0.36), (1.44±0.32), (1.39 ± 0.32) mm in 72 h, which were significantly decreased than 24 h admitted to ICU ( t values were2.029-4.504, P<0.05 or 0.01). During 1 week admission to ICU , the muscle fiber thickness showed a continuous trend of decline, and the muscle fiber thickness of the left biceps, right biceps, left gastrocnemius and right gastrocnemius decreased by 8.38% (0.13/1.55), 10.19% (0.16/1.57), 9.87% (0.15/1.52), 11.11% (0.17/1.53), respectively compared with the ICU admission. The muscle fiber thickness of the left biceps, right biceps, left gastrocnemius and right gastrocnemius decreased by 9.87% (0.15/1.52), 9.33% (0.14/1.50), 9.15% (0.14/1.53), 11.26% (0.17/1.51), respectively. Conclusion:Muscle attenuation can be observed within 48h after ICU entry, and it tends to decrease with the extension of the length of hospital stay.

Objective To investigate the expression of cystein rich 61 (Cyr61) and vascular endothelial growth factor (VEGF) in different stages of multiple myeloma (MM) patients,evaluate their relationship with angiogenesis and prognosis,and to determine the relationship between Cyr61 and VEGF.Methods Expression of Cyr61 and VEGF in BMMNC from 31 patients with different stages of MM and 10 cases of control were detected by RT-PCR.Results Expression of Cyr61 and VEGF in patients with MM (0.3652±1.5423,0.4815±1.3423) were significantly elevated in comparison to control (0.1862±0.7542,0.2012±1.2331) (P ＜ 0.05,P ＜ 0.01).Expression of Cyr61 and VEGF in stage Ⅲ (0.4632±0.1634,0.5356± 0.2342) was significantly higher than those in stage Ⅰ and stage Ⅱ (t =2.423,2.524,P ＜ 0.05),but there was no difference between stage Ⅰ and stage Ⅱ (0.2513±0.1365,0.3064±0.2124; 0.3084±0.2254,0.3653±0.1265) (t =1.782,1.824,P ＞ 0.05).The levels of Cyr61 and VEGF were significantly decreased after chemotherapy compared to before chemotherapy (P ＜ 0.01).Expression of Cyr61 and VEGF were positively correlated (r =0.8941,P ＜ 0.01).Conclusion Cyr61 and VEGF may play roles in the angiogenesis and pathophysiology of MM.It can be used to guide treatment and estimate prognosis by monitoring Cyr61 and VEGF.