BACKGROUND:Anaplastic thyroid carcinoma (ATC) is a rare but the most aggressive type of thyroid carcinoma. Nevertheless, limited advances were made to reduce mortality and improve survival over the last decades. Therefore, identifying novel diagnostic biomarkers and therapeutic targets for ATC patients is still needed. MATERIALS AND METHODS:RNA sequencing data and corresponding clinical features were available from GEO and TCGA databases. We integrated WGCNA and PPI network analysis to identify hub genes associated with ATC development, and RT-qPCR was employed for data verification. Univariate and LASSO Cox regression analyses were used to generate prognostic signatures. RESULTS:Based on PPI and WGCNA, 6 hub genes were identified, namely KIF2C, PBK, TOP2A, CDK1, KIF20A, and ASPM, which play vital roles in ATC development. Subsequently, RT-qPCR experiments showed that most of these genes were significantly upregulated in CAL-62 cells compared to Nthy-ori 3-1 cells. Moreover, a prognostic signature featuring GPSM2, FGF5, ASXL3, CYP4B1, CLMP, and DUXAP9 was generated, which was also verified by RT-qPCR results and proved as an independent predictor of poorer prognosis of ATC. Additionally, a nomogram incorporating the risk score and clinicopathological parameters was further constructed for accurate prediction of 1-, 3- and 5-year survival probabilities of ATC. CONCLUSIONS:Our study identified 6 key genes critical to ATC development and constructed a prognostic signature. These findings provide reliable biomarkers and a relatively comprehensive tumorigenesis profile of ATC, which may inform future strategies for clinical diagnosis and pharmaceutical design.

Defective renal excretion and increased production of uric acid engender hyperuricemia that predisposes to gout. However, molecular mechanisms underlying defective uric acid excretion remain largely unknown. Here, we report a rare genetic variant of gout-unprecedented NUMB gene within a hereditary human gout family, which was identified by an unbiased genome-wide sequencing approach. This dysfunctional missense variant within the conserved region of the NUMB gene (NUMBR630H) underwent intracellular redistribution and degradation through an autophagy-dependent mechanism. Mechanistically, we identified the uric acid transporter, ATP Binding Cassette Subfamily G Member 2 (ABCG2), as a novel NUMB-binding protein through its intracellular YxNxxF motif. In polarized renal tubular epithelial cells (RTECs), NUMB promoted ABCG2 trafficking towards the apical plasma membrane. Genetic loss-of-function of NUMB resulted in redistribution of ABCG2 in the basolateral domain and ultimately defective excretion of uric acid. To recapitulate the clinical situation in human gout patients, we generated a NUMBR630H knock-in mouse strain, which showed marked increases of serum urate and decreased uric acid excretion. The NUMBR630H knock-in mice exhibited clinically relevant hyperuricemia. In summary, we have uncovered a novel NUMB-mediated mechanism of uric acid excretion and a functional missense variant of NUMB in humans, which causes hyperuricemia and gout.

Objective The aim of the present study is to investigate the relationship between pulmonary hamartomas (PHs) and bronchi on multislice spiral computed tomography (MSCT) images. Methods The MSCT scans of 218 PHs from 216 pathologically confirmed patients were reviewed. The PHs were divided into two groups, namely, the central endobronchial and intraparenchymal groups, in accordance with location. Multiplanar reconstruction was used to demonstrate PH–bronchus relationship patterns. The PH–bronchus relationships in the intraparenchymal group were classified into five patterns: type I, the bronchus was cut off by the tumor; type II, the bronchus was contained within the tumor (air bronchogram sign); type III, the bronchus ran at the tumor periphery or was compressed by the tumor; and type IV, no tumor–bronchus relationship was observed. Results Nine (4.1%) PHs were assigned to the central endobronchial group and 209 (95.9%) PHs were assigned to the intraparenchymal group. In the endobronchial group, 1 (11.1%) PH was located in the trachea with the partial stenosis of the trachea, whereas the remaining 8 (88.9%) PHs were located in the lobar or segmental bronchus with the complete occlusion of the corresponding bronchus. In the intraparenchymal group, type IV (147, 70.3%) was most common pattern, followed by type III (54, 25.8%). Type I (8, 3.8%) was rare, and type II was not observed. Conclusion Central endobronchial PHs often obstructed bronchi, whereas only a few intraparenchymal PHs cut off bronchi. No air bronchogram sign was observed.

ObjectivesThis study aimed to distinguish preoperatively anterior mediastinal thymic cysts from thymic epithelial tumors via a computed tomography (CT)-based radiomics nomogram.MethodsThis study analyzed 74 samples of thymic cysts and 116 samples of thymic epithelial tumors as confirmed by pathology examination that were collected from January 2014 to December 2020. Among the patients, 151 cases (scanned at CT 1) were selected as the training cohort, and 39 cases (scanned at CT 2 and 3) served as the validation cohort. Radiomics features were extracted from pre-contrast CT images. Key features were selected by SelectKBest and least absolute shrinkage and selection operator and then used to build a radiomics signature (Rad-score). The radiomics nomogram developed herein via multivariate logistic regression analysis incorporated clinical factors, conventional CT findings, and Rad-score. Its performance in distinguishing the samples of thymic cysts from those of thymic epithelial tumors was assessed via discrimination, calibration curve, and decision curve analysis (DCA).ResultsThe radiomics nomogram, which incorporated 16 radiomics features and 3 conventional CT findings, including lesion edge, lobulation, and CT value, performed better than Rad-score, conventional CT model, and the clinical judgment by radiologists in distinguishing thymic cysts from thymic epithelial tumors. The area under the receiver operating characteristic (ROC) curve of the nomogram was 0.980 [95% confidence interval (CI), 0.963–0.993] in the training cohort and 0.992 (95% CI, 0.969–1.000) in the validation cohort. The calibration curve and the results of DCA indicated that the nomogram has good consistency and valuable clinical utility.ConclusionThe CT-based radiomics nomogram presented herein may serve as an effective and convenient tool for differentiating thymic cysts from thymic epithelial tumors. Thus, it may aid in clinical decision-making.

Ribosomal S6 kinase 4 (RSK4) is a putative tumor suppressor gene which is inactivated by epigenetic events in a number human malignancies; however, its role in papillary thyroid carcinoma (PTC) remains largely unknown. The aim of this study was to explore the methylation status of the RSK4 promoter in PTC, and to determine its potential role in thyroid carcinogenesis. Reverse transcription‑quantitative PCR (RT‑qPCR) and western blot analyses were performed to examine the RSK4 mRNA and protein levels, respectively. Methylation‑specific PCR (MSP) and bisulfite genomic sequencing (BGS) were used to analyze methylation status of the RSK4 gene. Sanger sequencing was further carried out to detect the BRAF V600E mutation. Cell proliferation assay was finally performed to evaluate the role of hypermethylation in the growth of PTC cells. The association between RSK4 methylation and the clinicopathological characteristics of patients with PTC was assessed. The methylation frequency of the RSK4 promoter in PTC tissues was higher than that in paired paracancerous tissues. Coincidentally, the RSK4 mRNA levels were also downregulated in PTC tissues when compared with the paracancerous counterparts. The hypermethylation of RSK4 was associated with tumor size and lymph node metastasis. Furthermore, the BRAF V600E mutation may influence RSK4 expression and methylation. Moreover, RSK4 hypermethylation was observed in thyroid cancer cell lines, which was consistent with a lack of RSK4 expression. Upon the 5‑Aza‑deoxycytidine treatment of thyroid cancer cells, RSK4 expression was significantly upregulated, while cell proliferation was inhibited. On the whole, the findings of the present study demonstrate that the hypermethylation of the RSK4 promoter may be one of the mechanisms responsible for the poor RSK4 expression in PTC. Thus, these data suggest that RSK4 may serve as a molecular target for the early diagnosis and treatment of PTC.

Objective In recent years,the incidence of papillary thyroid carcinoma(PTC)increased gradually and the recurrence and metastasis of PTC patients are also increasing.The aim of this study was to investigate the effect of BRAFV600E mutation on the expression of HMGB1 in PTC,to explore the mechanism of BRAF gene affecting the development and prognosis of PTC,and then guiding clinical practice of precise therapy.Methods The preoperative serum and postoperative fresh tissue were collected from 44 cases of PTC patients in our hospital from September,2015 to December,2015.According to BRAFV600E mutations divided into positive group and negative group.Western blot,immunohistochemistry and fluorescence quantitative PCR were used to detect the level of HMGB1 in cancer tissues.The level of HMGB1 in serum was tested by ELISA method.Western blot was performed using Image J software.The relative quantitative method was used to calculate the fluorescence quantitative PCR data.The concentration of HMGB1 protein in serum was calculated by ELISA Calc regression/fit calculation program.All the data were analyzed by SPSS 20.0.Western blot,fluorescence quantitative PCR and ELISA data were analyzed by Chi-square test,Mann-Whitney-Wilcoxon test and independent sample t test.The relationship between lymph node metastasis and extrathyroid infiltration and the occurrence of BRAFV600E mutation was analyzed by chi-square test.Results In PTC cancer tissues,HMGB1 was mainly located in the cytoplasm,the levels of HMGB1mRNA and protein in positive group were lower than that of the negative group(Z =2.117,P ＜0.01,x2 =19.989,P ＜ 0.05),and the change did not show in peripheral blood(t =1.135,P ＞ 0.05).The expression of HMGB1 mRNA and protein in PTC with lymph node metastasis was lower than those without lymph node metastasis(Z =-2.216,P ＜0.05;t =-2.217,P ＜ 0.05),and so as in the extrathyroid infiltration(Z =-2.267,P ＜ 0.05;t =-3.885,P ＜ 0.01).Conclusion BRAFV600E mutations may accelerate the malignant development of PTC by down regulating the expression of HMGB1.

BACKGROUND:Papillary thyroid cancer (PTC) is the most common differentiated thyroid cancer and is responsible for 80-90% of thyroid cancer cases. Despite typically excellent prognoses, these subclinical low-risk cancers are often treated aggressively by surgical thyroidectomy. Consequently, the objective of this study was to generate a prognostic matrix to be used prior to PTC intervention.METHODS:In this study, 80 PTC patients were assessed. Following adjustment for sex, logistic regression analysis showed that BRAFV600E mutation, transforming growth factor beta (TGF-β) expression, age, and tumor size are risk factors that can affect tumor clinical stage (p < 0.05). Based on the results of this analysis, we generated a matrix that incorporated 4 variables: patient age, tumor size, BRAFV600E mutation, and TGF-β expression.RESULTS:We observed that the corresponding area under curve was as high as 0.91. The sensitivity and specificity of the model were 94.74 and 83.61%, respectively. These values are significantly higher than those generated from single indexes.CONCLUSION:As a result of this analysis, it is hoped that the resultant matrix can be utilized during clinical diagnosis and treatment prior to thyroid nodule surgery.

Breast cancer was a malignant tumor seriously threatening the life of women in the world. But the prognosis of breast cancer patients was not so satisfactory due to the limited effective therapeutics. The heterogeneity decided that more than one gene or one signaling pathway was responsible for the initiation or progression of breast cancer. DDX51 gene was a member of RNA helicases family in charge of regulation of RNA metabolism. And DDX51 gene was shown to promote proliferation in NSCLC. But we firstly reported the abundant expression of DDX51 gene in both the breast cancer tissues and cell lines in this study. And DDX51 expression was shown to be associated with TNM stage and prognosis in breast cancer patients. When DDX51 was successfully knocked down, either proliferation or DNA synthesis of MCF-7 cells was inhibited. But the ability of migration and invasion of MCF-7 cells was not affected by DDX51 gene. Furthermore, DDX51 knockdown was accompanied by inhibition of Wnt/β-catenin signaling because expression of critical members such as β-catenin, cyclin D1, TCF/LEF, and DKK1 were all affected. Therefore, this study proved that DDX51 gene promoted proliferation in MCF-7 cells by regulating Wnt/β-catenin signaling pathway and showed clinical significance in breast cancer. This study provides us a new promising hope for treatment of patients with breast cancer.

A poor papillary thyroid cancer (PTC) prognosis is strongly associated with the BRAF V600E mutation. During tumor progression, levels of the high mobility group box 1 (HMGB1) protein are often dysregulated. Results herein demonstrate that HMGB1 protein levels differ between tumor and adjacent non-tumor tissue and that HMGB1 mRNA levels were higher in wild-type BRAF PTC tissues than in BRAF V600E PTC tissues (2-△Ct 0.31 ± 0.25 vs. 0.16 ± 0.12; P < 0.05). HMGB1 protein levels also differed in the same manner (wild-type BRAF PTC tissues 0.11 ± 0.04 vs. BRAF V600E PTC tissues 0.03 ± 0.03; P < 0.001). Although not detected in peripheral blood, low levels of HMGB1 were significantly related to PTC cell lymph node metastasis and extra-glandular infiltration (P = 0.045 and P = 0.002). Experimental results at the cellular level were consistent with tissues and further verified the relationship of BRAF V600E and HMGB1. These findings demonstrate that the BRAF V600E mutation down-regulates levels of HMGB1, likely through activation of the mitogen-activated protein kinase (MAPK) signaling pathways.

Lipoprotein lipase (LPL) is widely expressed in skeletal muscles, cardiac muscles as well as adipose tissue and involved in the catabolism of triglyceride. Herein we have systematically characterized two novel loss-of-function mutations in LPL from a Chinese family in which afflicted members were manifested by severe hypertriglyceridemia and recurrent pancreatitis. DNA sequencing revealed that the proband was a heterozygote carrying a novel c.T928C (p.C310R) mutation in exon 6 of the LPL gene. Another member of the family was detected to be a compound heterozygote who along with the c.T928C mutation also carried a novel missense mutation c.A1187T (p.E396V) in exon 8 of the LPL gene. Furthermore, COS-1 cells were transfected with lentiviruses containing the mutant LPL genes. While C310R markedly reduced the overall LPL protein level, COS-1 cells carrying E396V or double mutations contained similar overall LPL protein levels to the wild-type. The specific activity of the LPL mutants remained at comparable magnitude to the wild-type. However, few LPL were detected in the culture medium for the mutants, suggesting that both mutations caused aberrant triglyceride catabolism. More specifically, E396V and double mutations dampened the transport of LPL to the cell surface, while for the C310R mutation, reducing LPL protein level might be involved. By characterizing these two novel LPL mutations, this study has expanded our understanding on the pathogenesis of familial hypertriglyceridemia (FHTG).

DACH1 has been found down-regulated in a variety of human cancers, but its clinical significance and functional roles in colorectal cancer (CRC) remain unknown. In this study, we identified DACH1 as a tumor suppressor in CRC. Suppression of DACH1 strikingly increased cell growth, migration and invasion potential of CRC cell line SW480. Expression analysis of a set of epithelial-mesenchymal transition (EMT) markers by RT-qPCR and western blot showed an increase in the expression of mesenchymal markers (vimentin and N-cadherin) and a reduction in the expression of epithelial marker (E-cadherin and γ-catenin). Furthermore, EMT characteristics in DACH1-downregulated CRC cells were abrogated by TGF-β inhibitor SB431542. DACH1 overexpression reduced TGF-β-induced EMT and inhibited SW480 cell invasion which can be reversed in the presence of TGF-β. Thus, our results suggest that DACH1 loss of function results in increased cell growth, motility and invasiveness through TGF-β-mediated EMT, and DACH1 loss of function has important therapeutic implications for targeted therapies of CRC.

Objective To investigate the prevalence of BRAF T1799A mutation and RET/PTC rearrangement in Qingdao and detect the expression of platelet-derived growth factor B（PDGF-B） in order to investigate the correlation

Objective: To estimate the prevalence and risk factors of hypertension in males over the age of 20 in Shandong coastal areas in 2009, and compare the results to that of 2004.Methods: A total of 2,235 male inhabitants from Qingdao, Yantai, Weihai, Rizhao and Dongying selected by random stratified cluster sampling was investigated through household surveys. Results:The standardized prevalence rate of male hypertension significantly increased from 32.1% in 2004 to 39.6% in 2009 (P<0.01).The prevalence of hypertension in male increased with age, and the highest was among individuals over 70 years old (P<0.01).The prevalence of individuals over 50 years old was higher than the younger groups (P<0.01).In contrast to 2004, the average age, BMI, waist circumference, triglyceride, total cholesterol, uric acid and blood glucose levels were significantly increased in 2009 (P<0.01).Logistic regression analysis showed that age, BMI, waist circumference, lipid disorders, uric acid, blood glucose and family history were risk factors in male hypertension. Conclusion:The prevalence of hypertension in male was significantly increased in 2009.Aging, overweight or obesity, hyperglycemia, hyperuricemia, lipid disorders and family history were independent risk factors of hypertension.