[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.

[Application of enhanced recovery after surgery in laryngeal cancer surgery with multi-disciplinary team].

Objective: To explore the application value of enhanced recovery after surgery (ERAS) with the multidisciplinary team (MDT) model in laryngeal cancer surgery. Methods: Eighty patients with laryngeal cancer treated in Department of Otorhinolaryngology Head and Neck Surgery of Yantai Yuhuangding Hospital from May 2016 to June 2017 were selected, including 76 males and 4 females, aged 45 to 75 years old. By random number table method, they were divided into ERAS group (40 cases) and control group (40 cases). Visual analogue scale (VAS), general comfort questionnaire (GCQ) and self-rating Anxiety Scale (SAS) were used to evaluate the symptoms and signs and psychological state of the two groups before and after operation. Mann Whitney U test was used for non-normal distribution data, and chi square test, Fisher exact probability method and covariance analysis were used for classification data. Repeated measures analysis of variance was used for the comparison of each group at different time points. Results: Two cases in the ERAS group and six cases in the control group withdrew from the study for some reason. Finally, 38 cases in the ERAS group and 34 cases in the control group were enrolled in this study. The postoperative pain scores of the two groups were the highest at 6 h after operation, and then gradually decreased. At different time points after operation, the pain scores of ERAS group were lower than those of the control group. At 24 h after operation, the pain relief degree of ERAS group was significantly higher than that of the control group, with a statistically significant difference (P<0.05). Compared to control group, ERAS group had lower preoperative thirst score [(0.15±0.36) vs. (4.29±1.17), Z=-7.695, P<0.001] and hunger score [(0.38±0.49) vs. (3.44±1.13), Z=-7.426, P<0.001]. The total number of postoperative adverse reactions (8 vs.16), oral feeding time [(4.06±4.42) d vs. (9.06±2.42) d] and postoperative hospital stay [(5.91±0.97) d vs. (11.03±2.11)d] in ERAS group were lower than those in control group (statistics 5.461, -4.558, -7.347, P<0.05), but there was no significant difference in postoperative catheter indwelling time and neck drainage tube indwelling time between the two groups (P>0.05). Before discharge, the comfort of ERAS group was significantly higher than that of control group [(60.37±8.78) vs. (50.38±8.08), Z=-4.370, P<0.001]. Before discharge, the anxiety level of ERAS group decreased, while that of the control group increased significantly, which was higher than that of ERAS Group [(59.12±6.43) vs. (52.62±6.25), Z=-4.179, P<0.001]. Conclusion: The application of multidisciplinary ERAS in laryngeal cancer surgery can improve preoperative hunger and thirst, postoperative pain and mental state, shorten the length of hospital stay and reduce postoperative adverse reactions.

[Radiomics nomogram of MR: a prediction of cervical lymph node metastasis in laryngeal cancer].

Objective: To establish and validate a radiomics nomogram based on MR for predicting cervical lymph node metastasis in laryngeal cancer. Methods: One hundred and seventeen patients with laryngeal cancer who underwent MR examinations and received open surgery and neck dissection between January 2016 and December 2019 were included in this study. All patients were randomly divided into a training cohort (n=89) and test cohort (n=28) using computer-generated random numbers. Clinical characteristics and MR were collected. Radiological features were extracted from the MR images. Enhanced T1 and T2WI were selected for radiomics analysis, and the volume of interest was manually segmented from the Huiyihuiying radiomics cloud platform. The variance analysis (ANOVA) and the least absolute shrinkage and selection operator (LASSO) algorithm were used to reduce the dimensionality of the radiomics features in the training cohort. Then, a radiomic signature was established. The clinical risk factors were screened by using ANOVA and multivariate logistic regression. A nomogram was generated using clinical risk factors and the radiomic signature. The calibration curve and receiver operator characteristic (ROC) curve were used to confirm the nomogram's performance in the training and test sets. The clinical usefulness of the nomogram was evaluated by decision curve analysis (DCA). Furthermore, a testing cohort was used to validate the model. Results: The radiomics signature consisted of 21 features, and the nomogram model included the radiomics signature and the MR-reported lymph node status. The model showed good calibration and discrimination. The model yielded areas under the ROC curve (AUC) in the training cohort, specificity, and sensitivity of 0.930, 0.930 and 0.875. In the test cohort, the model yielded AUC, specificity and sensitivity of 0.883, 0.889 and 0.800. DCA indicated that the nomogram model was clinically useful. Conclusion: The MR-based radiomics nomogram model may be used to predict cervical lymph node metastasis of laryngeal cancer preoperatively. MR-based radiomics could serve as a potential tool to help clinicians make an optimal clinical decision.

[Clinical features and gene mutation analysis of 13 Chinese juvenile patients with nephronophthisis].

Objective: To explore the clinical features and pathogenic gene mutation of juvenile nephronophthisis (NPHP) in Chinese patients. Method: Clinical data and blood samples of 27 juvenile NPHP patients from 25 families who were initially clinically diagnosed in six hospitals in Guangdong province were collected. NPHP1 homozygous deletions were detected in all patients. Sequencing of NPHP1 gene was performed when homozygous deletions were not found in patients without eye involvement. In patients with eye involvement, NPHP5 sequencing was carried out initially and subsequently NPHP10 gene and NPHP1 when there were no NPHP5 gene mutation found. Result: Diagnosis was confirmed in 13 patients by renal pathology and (or) gene sequencing, including four boys and nine girls with a median onset age of 8.5(0.1-12.8) years. Seven of the 13 patients had a normal routine urine test and six patients had mild to moderate proteinuria. None had persistent hematuria. The estimated glomerular filtration rate of the 13 patients was (12.7±10.7) ml/(min·1.73 m2) at the time of diagnosis. Renal cysts were found in only five patients by iconography. Decreased renal size was observed in nine cases and normal renal size in four patients. Renal pathology was available in five patients, renal cysts formation at the cortical-medullar area, thickening and laying tubular basement membrane, were observed. Two of the thirteen children had eye involvement, one had liver impairment and one had growth retardation. NPHP1 gene defects were detected in seven patients with a mutation rate of 25.9%, and large homozygous deletions were observed in three patients. Four patients had single point mutations, i. e. compound heterozygous mutations (c.13 C>T and c. 1520+ 5 G>A) in one patient; homozygous mutation in three patients, two patients were siblings from the same pedigree harbored c. 1756 C>T and the other one harbored c. 1298delA. NPHP5 gene homozygous mutation was found in one pedigree. The fourteen children without renal pathology and whose genetic tests were negative shared similar clinical features with the thirteen patients whose diagnosis were confirmed by gene mutation and (or) renal pathology. Conclusion: The onset of juvenile NPHP is insidious. Urine and renal iconography changes are mild or negative. The ratio of NPHP1 mutant patient is similar with previous reports, but the proportion of NPHP1 gene homozygous deletions is much lower and all of the NPHP1 gene single point mutations detected in this research were novel, which indicates a genetic discrepancy existed between Chinese NPHP patients and the western ones.