[Correlation analysis of clock genes and MEN2 medullary thyroid carcinoma].

Objective: To investigate the correlation between CLOCK and BMAL1 genes and MEN2 medullary thyroid carcinoma (MTC). Methods: Thirteen cases with MEN2 MTC and thirteen cases with non-MEN2 MTC were selected who were treated in the Yantai Yuhuangding Hospital between January 2013 and September 2021. Clinical indicators such as blood calcitonin level, tumor diameter and metastatic lymph node of patients were collected. The expression differences of CLOCK and BMAL1 between MEN2 MTC and para-carcinoma tissue as well as between MEN2 MTC and non-MEN2 MTC were detected by immunohistochemistry and qPCR. The correlation between lymph node metastasis and CLOCK or BMAL1 expression was analyzed. Protein-protein interaction (PPI) network analysis combined with qPCR and correlation analysis was used to explore the expression regulation relationship between RET and circadian clock genes. The rhythm disorder of MEN2 cells was verified by lipopolysaccharide cell stimulation experiment after dexamethasone rhythm synchronization. Results: MEN2 MTC exhibited typical RET gene mutation. The mean blood calcitonin level, the tumor diameter and the number of metastatic lymph nodes of patients with MEN2 MTC were higher than those of patients with non-MEN2 MTC (t value was 2.76, 2.53, 2.26, all P<0.05). Immunohistochemical results showed that the expression levels of CLOCK and BMAL1 in MEN2 MTC were higher than those in non-MEN2 MTC, while negatively expressed in para-cancerous thyroid follicle. qPCR displayed that the expression of CLOCK gene in cancer tissues was higher than that in non-MEN2 MTC and para-cancerous tissues (t value was 2.68 and 2.86, all P<0.05); the expression of BMAL1 gene in MEN2 MTC was higher than that in non-MEN2 MTC and para-cancerous tissues (t value was 2.21 and 2.35, all P<0.05). Correlation analysis showed that the expression levels of CLOCK and BMAL1 genes were positively correlated with the number of lymph node metastases in patients with MEN2 MTC (r=0.65, P<0.001; r=0.52, P=0.005). PPI network analysis indicated that the expression of CLOCK gene was positively correlated with the abnormal expression of RET gene (r=0.96, P<0.001). With lipopolysaccharide to stimulate cultured cells in vitro after dexamethasone rhythm synchronization, the expressions of CLOCK and BMAL1 in MEN2 MTC cells (0.47±0.22 and 2.60±1.48) at 12 hours of synchronization were significantly lower than those in para-cancerous tissues (1.70±1.62 and 8.23±2.52), the difference was statistically significant(t=5.04, P=0.007; t=3.34, P=0.029). Conclusion: CLOCK and BMAL1 are correlated with the occurrence and development of MEN2 MTC, and may be potential targets for the development of new therapeutic strategies for MEN2 MTC.

LINC00106 prevents against metastasis of thyroid cancer by inhibiting epithelial-mesenchymal transition.

OBJECTIVE: Thyroid cancer (TC) is a common malignant tumor of the endocrine system, and its morbidity and mortality are in the high places. Recent studies have focused on exploring biological markers and targeted therapy for TC. This research aims to elucidate the role of LINC00106 in the progression of TC and the regulatory mechanisms. PATIENTS AND METHODS: Differential level of LINC00106 in a downloaded profile containing TC and normal tissues from GEPIA database was analyzed. Subsequently, its level in TC tissues and cell lines was detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The relationship between LINC00106 level and clinical data of TC patients was assessed, including age, tumor staging, lymphatic metastasis, and overall survival. After transfection of si-LINC00106, TC cell metastasis was evaluated by wound healing and transwell assay. Relative levels of E-cadherin, N-cadherin, ß-catenin, and Vimentin regulated by LINC00106 were determined using qRT-PCR and Western blot. RESULTS: LINC00106 was downregulated in TC tissues than normal ones. Its level was correlated to tumor staging, lymphatic metastasis and overall survival in TC patients. The knockdown of LINC00106 in BCPCP and TPC-1 cells enhanced migratory and invasive abilities and triggered the process of epithelial-mesenchymal transition (EMT). CONCLUSIONS: LINC00106 is lowly expressed in TC specimens, which attenuates migratory and invasive abilities in TC by inhibiting EMT as a tumor suppressor.

[Clinical significance of lymphatic metastasis between sternocleidomastoid and sternohyoid muscle in papillary thyroid cancer].

Objective: To explore the clinical significance of metastasis of lymph nodes between sternocleidomastoid and sternohyoid muscle (LNSS) in papillary thyroid cancer (PTC). Methods: A total of 175 patients with PTC who underwent thyroidectomy with LNSS dissection were retrospectively analyzed. Univariate and multivariate Logistic regression analyses were used to determine the independent risk factors for LNSS metastasis in PTC. Results: The rate of detectable LNSS was 70.9% (124/175) and metastasis rate was 7.4% (13/175). Of 13 cases with LNSS metastasis, 10 with the coexistence of cervical lymph node metastasis. Univariate Logistic regression analysis showed that multiple focal cancer, tumor located in the lower pole of thyroid, belt-shaped muscle invasion, lateral cervical lymph node metastasis, cN+ , the number of cervical lymph nodes with metastasis and the number of lymph nodes with metastasis in level Ⅳwere the risk factors for LNSS metastasis (P<0.05). Multivariate Logistic regression analysis suggested that tumor located in the lower pole of thyroid and the number of cervical lymph nodes with metastasis >6 were the independent risk factors for LNSS metastasis (P<0.05). Given the number of cervical lymph nodes with metastasis as a predictor for the LNSS metastasis, the sensitivity was 92.3%, the specificity was 66.7% and the accuracy rate was 68.6%. Conclusions: LNSS metastasis is commom in PTC, with a metastasis rate of 7.4%. PTC in the lower pole of thyroid and the number of cervical lymph nodes with metastasis > 6 are independent risks for LNSS metastasis.

[The clinical significance of Delphian lymph node metastasis in papillary thyroid cancer].

Objective: To evaluate the clinical significance of Delphian lymph node (DLN) metastasis in papillary thyroid cancer (PTC). Method: A total of 505 cases with PTC confirmed pathologically in our hospital between January 2015 and December 2015 were retrospectively reviewed. 208 patients with DLN assessed separately by histopathologic examination who underwent primary surgery for PTC were included for the following analysis. Results: In 208 patients, the detection rate of DLN was 63.0% and the metastasis rate of DLN was 21.4%. DLN metastasis was correlated with PTC multifocality (P=0.038), tumor size over 1cm (P=0.001), BRAFV600E mutation (P=0.017) and central neck node metastasis (P<0.001). Tumor size over 1cm (95%CI 1.308-9.909, OR=3.600, P=0.013) and the number of node with central neck metastasis (95%CI 1.313-2.163, OR=1.685, P<0.001) were independent risk factors for DLN metastasis. The presence of DLN metastasis was associated with an 8.8-fold higher frequency of central neck node metastasis compared to cases without DLN metastasis. Among patients with DLN metastases, central lymph node metastasis was more common in the cases with lateral neck node metastases compared to those without lateral neck node metastases (6.5±3.0 vs 1.5±0.7, P=0.009), and 5 of the 6 patients also presented with PTC multifocality and BRAFV600E mutation. Conclusion: DLN metastasis implies a higher possibility of central neck lymph node metastasis. DLN should be assessed during operation to provide information for neck dissection, post-operative administration and follow-up strategy.

Study on serum proteomic features in patients with and without recurrence or metastasis after surgical resection of esophageal carcinoma.

The purpose of this study was to identify specific bio-markers for recurrence or metastasis of esophageal carcinoma in serum of patients subjected to esophagectomy. Surface-enhanced laser desorp-tion/ionization time-of-flight mass spectrometry (SELDI-TOF-MS) combined with IMAC-Cu(2+) ProteinChip array were performed for the serum protein profiling in patients after surgical resection of esophageal carcinoma. Two groups of patients were analyzed: 38 patients without recurrence or metastasis (Group 1) and 22 patients with recurrence or metastasis after resection (Group 2). The Biomarker Wizard and Bio-marker Patterns software were used to identify proteins differentially expressed between the 2 groups. There were 33 differentially expressed serum proteins detected by comparison between the groups. The clas-sification tree model composed of 3 differentially expressed proteins with different m/z (9368.63, 5342.59, and 5254.43 Da) was established. Under the learning mode, the sensitivity and specificity of this model for diagnosis of esophageal carcinoma recurrence or metastasis were both 100% (22/22 and 38/38, respectively). Under the testing mode, the sensitivity and specificity were 90.9% (20/22) and 94.7% (36/38), re-spectively. The recurrence or metastasis of esophageal carcinoma after esophagectomy can be rapidly and accurately detected using the combi-nation of SELDI-TOF-MS with IMAC-Cu(2+) ProteinChip array, which, therefore, has a potential for clinical application.

Filariasis and its control in Fujian, China.

Epidemiological survey of filariasis in Fujian Province, China showed that malayan filariasis, transmitted by Anopheles lesteri anthropophagus was mainly distributed in the northwest part and bancroftian filariasis with Culex quinquefasciatus as vector, in middle and south coastal regions. Both species of filariae showed typical nocturnal periodicity. Involvement of the extremities was not uncommon in malayan filariasis. In contrast, hydrocele was often present in bancroftian filariasis, in which limb impairment did not appear so frequently as in the former. Hetrazan treatment was administered to the microfilaremia cases identified during blood examination surveys, which were integrated with indoor residual spraying of insecticides in endemic areas of malayan filariasis when the vector mosquito was discovered and with mass treatment with hetrazan medicated salt in endemic areas of bancroftian filariasis. At the same time the habitation condition was improved. These factors facilitated the decrease in incidence. As a result malayan and bancroftian filariasis were proclaimed to have reached the criterion of basic elimination in 1985 and 1987 respectively. Surveillance was pursued thereafter and no signs of resurgence appeared.