BRAF-activated WT1 contributes to cancer growth and regulates autophagy and apoptosis in papillary thyroid carcinoma.

BACKGROUND: Papillary thyroid carcinoma (PTC) is one of most prevalent malignant endocrine neoplasms, and it is associated with a high frequency of BRAF gene mutations, which lead to lymphatic metastasis and distant metastasis that promote tumor progression. The molecular mechanism of PTC and the role of BRAF mutation in PTC progression and development need to be further elucidated. METHODS: In this study, a comprehensive bioinformatics analysis was performed to identify the differentially expressed genes and signaling pathways in thyroid cancer patients carrying mutant BRAF. Then, we confirmed the prognostic role of WT1 in thyroid cancer patients. Immunohistochemistry was performed to measure the expression profile of WT1 in PTC tissue. Lentivirus shWT1 was transfected into BRAFV600E (mutant) PTC cells to stably inhibit WT1 expression. CCK-8, EdU, immunofluorescence, colony formation, cell migration, cell wound healing, apoptosis and autophagy assays were performed to assess the biological functions of WT1 in BRAFV600E PTC cells. RNA sequencing, immunohistochemistry and immunoblotting were performed to explore the molecular mechanism of WT1 in BRAFV600E PTC cells. RESULTS: The results confirmed that "epithelial cell proliferation", "apoptosis" and "selective autophagy" were closely associated with this BRAF mutant in these thyroid cancer patients. Knocking down BRAF-activated WT1 effectively inhibited the proliferation and migration of BRAFV600E PTC cells. Silencing WT1 significantly inhibited autophagy and promoted the apoptosis of BRAFV600E PTC cells. Mechanistic investigations showed that silencing WT1 expression remarkably suppressed the AKT/mTOR and ERK/P65 signaling pathways in BRAFV600E PTC cells. CONCLUSION: All these results indicate that WT1 is a promising prognostic biomarker and facilitates PTC progression and development of cells carrying the BRAFV600E mutation.

Feasibility of using colloidal gold immunochromatography for point-of-care identification of parathyroid glands during thyroidectomy.

AIM: To determine the feasibility of using colloidal gold immunochromatography for rapid identification of parathyroid glands during thyroidectomy. MATERIAL AND METHODS: 127 patients undergoing thyroidectomy were randomly divided into PTH-ICGT group (64 cases) and conventional naked eye group (63 cases). The rate of identification of parathyroid glands and the incidence of hypoparathyroidism were compared between the two groups. RESULTS: PTH-CGI assay results showed that PTH concentration in the parathyroid tissue was (955.3 ±â€¯16.1) ng/L; skeletal muscle tissue [(14.5 ±â€¯1.5) ng/L], thyroid tissue [(15.0 ±â€¯1.3) ng/L], adipose tissue [(15.3 ±â€¯1.2) ng/L], lymph node tissue [(14.0 ±â€¯1.2) ng/L];PTH levels in parathyroid tissues were compared with PTH levels in skeletal muscle, thyroid, fat, and lymph node tissues, respectively. The differences were statistically significant(t values were 23.62, 33.42, 39.34, 30.77, P < 0.0001, respectively); Among the 127 patients undergoing total thyroidectomy, the rate of detection of parathyroid glands was 92.7% in the conventional naked eye group and 96.4% in the PTH-ICGT group. There was no significant difference in the detection rate of parathyroid gland between the two groups (χ2 = 0.7067, P = 0.40). The incidence of temporary hypoparathyroidism after surgery in both groups was 11.3% and 5.7%, respectively (χ2 = 1.093, P > 0.05). The incidence of postoperative permanent hypoparathyroidism in both groups was 3.8% and 0, respectively (Fisher's exact test, P = 0.495). CONCLUSION: PTH-CGI has a high efficiency in identifying parathyroid glands, which may increase the rate of clinical parathyroid detection and reduce the incidence of postoperative hypoparathyroidism.

Application of Preoperative Ultrasonography in the Diagnosis of Cervical Lymph Node Metastasis in Thyroid Papillary Carcinoma.

Background: The clinical value and application of preoperative ultrasound contrast in the diagnosis of cervical lymph node metastasis in thyroid papillary carcinoma is investigated. Methods: In total, 126 cases of thyroid papillary carcinoma were selected, the sensitivity and accuracy of color ultrasound and ultrasound contrast were analyzed by comparing preoperative gray-scale ultrasound, color ultrasound, and ultrasound contrast. Results: The accuracies of preoperative color ultrasound and ultrasound contrast in detecting lymph node metastasis were 74 and 82%, respectively, and their sensitivities were 80 and 94%, respectively. Lymph node metastasis was significantly more severe when the tumor diameter was >4 cm. The lymphatic metastatic rate of the patients with multifocal papillary carcinoma was 96.4%, whereas the lymphatic metastatic rate of the patients with thyroid gland lesions was 87.7%. The central foci of cervical lymph node metastasis included the following pathological subtypes: diffuse sclerosis type (89.3%, 25/28), high-cell type (72.2%, 8/11), and papillary type (40.0%, 4/10). Conclusion: Ultrasound contrast is more sensitive than color ultrasound in the diagnosis of cervical lymph node metastasis. Primary lesions ≥4 cm, lesion involvement, outer membrane, and high-risk pathologic subtypes and lesions were considered as the criteria for ultrasound contrast application.

Ultrasound-Guided Radiofrequency Ablation: A New Attempt to the Treatment of Refractory Hyperparathyroidism Secondary to Chronic Kidney Disease.

INTRODUCTION: To evaluate clinical application value of radiofrequency ablation (RFA) in refractory hyperparathyroidism secondary to chronic kidney disease (CKD) by comparing the safety and effectiveness of RFA with parathyroidectomy with autotransplantation (PTX + AT). METHODS: A retrospective study was conducted on 80 patients with CKD with refractory hyperparathyroidism who underwent RFA or PTX + AT between January 2018 and February 2021. Serum parathyroid hormone (PTH), calcium, and phosphorus levels, complications, clinical symptoms, visual analogue scale (VAS) scores, hospital stay duration, and postoperative recurrence rate were compared between the 2 groups. RESULTS: Serum PTH, phosphorous, and calcium levels and the VAS scores improved significantly after RFA and PTX + AT (P < 0.05). Significant differences were observed between the 2 groups in postoperative (day 1 and week 1) levels of serum PTH and postoperative day 1 of serum calcium and phosphorus levels (P < 0.05), with more pronounced reduction after PTX + AT. Although the incidence of recurrent laryngeal nerve (RLN) injury was slightly higher in the RFA group compared with the PTX + AT group (26.67% vs. 16.67%; P > 0.05), RFA markedly decreased the risk of severe hypocalcemia (SH) (20% vs. 46.67%; P < 0.05) and shortened hospital stay (7.53 ± 2.67 days vs. 12.13 ± 3.86 days; P < 0.05). The 6-month recurrence rate was 23.3% (7 of 30) in the RFA group and 30% (9 of 30) in the PTX + AT group (P > 0.05). CONCLUSION: RFA can treat refractory secondary hyperparathyroidism (SHPT) with similar clinical efficacy as surgery and achieve faster recovery and a lower incidence of postoperative SH.

Regulation of GSK-3 beta in the proliferation and apoptosis of human thyrocytes investigated using a GSK-3 beta-targeting RNAi adenovirus expression vector: involvement the Wnt/beta-catenin pathway.

Disorders in the proliferation and apoptosis of thyrocytes may induce goitre, adenoma and carcinoma in the thyroid. The Wnt/beta-catenin pathway has been demonstrated to be involved in the regulation of cell proliferation, differentiation and apoptosis in various cell lines. The regulatory mechanism on the proliferation and differentiation of thyrocytes is not well characterized. In the present study, a GSK-3beta-targeting RNA interference (RNAi) adenovirus vector was constructed and delivered to primary human thyrocytes. Results showed that the expression of beta-catenin protein in primary human thyrocytes was increased after GSK-3beta-targeting RNAi adenovirus infection, the proliferation of primary human thyrocytes was significantly stimulated using Bromodeoxyuridine (BrdU) assay, while cell apoptosis was slightly affected which was observed through flow cytometry. It is concluded that the Wnt/beta-catenin pathway plays a significant role in the regulation of the proliferation of primary human thyrocytes.

A six-gene-based signature for breast cancer radiotherapy sensitivity estimation.

Breast cancer (BRCA) represents the most common malignancy among women worldwide with high mortality. Radiotherapy is a prevalent therapeutic for BRCA that with heterogeneous effectiveness among patients. Here, we proposed to develop a gene expression-based signature for BRCA radiotherapy sensitivity estimation. Gene expression profiles of BRCA samples from the Cancer Genome Atlas (TCGA) and International Cancer Genome Consortium (ICGC) were obtained and used as training and independent testing dataset, respectively. Differential expression genes (DEGs) in BRCA samples compared with their paracancerous samples in the training set were identified by using the edgeR Bioconductor package. Univariate Cox regression analysis and LASSO Cox regression method were applied to screen optimal genes for constructing a radiotherapy sensitivity estimation signature. Nomogram combining independent prognostic factors was used to predict 1-, 3-, and 5-year OS of radiation-treated BRCA patients. Relative proportions of tumor infiltrating immune cells (TIICs) calculated by CIBERSORT and mRNA levels of key immune checkpoint receptors was adopted to explore the relation between the signature and tumor immune response. As a result, 603 DEGs were obtained in BRCA tumor samples, six of which were retained and used to construct the radiotherapy sensitivity prediction model. The signature was proved to be robust in both training and testing sets. In addition, the signature was closely related to the immune microenvironment of BRCA in the context of TIICs and immune checkpoint receptors' mRNA levels. In conclusion, the present study obtained a radiotherapy sensitivity estimation signature for BRCA, which should shed new light in clinical and experimental research.