Objective:To investigate the molecular mechanism of miR-886-5p targeting BCL-2-associated X protein (BAX) to inhibit the proliferation, migration, and invasion of liver cancer cells.Methods:mRNA expression data of HCC patients were obtained from the Starbase database, including 370 liver cancer samples and 50 normal liver tissue samples adjacent to the cancer. Analyze the expression of miR-886-5p in the previously obtained data and investigate the relationship between miR-886-5p and BAX in liver cancer samples. After transfection of the corresponding plasmids into Huh7 and HepG2 cells, the following groups were established. Analyze the interaction between miR-886-5p and BAX in vitro, detect the protein expression by Western blotting, and verify the targeting relationship between the two by dual luciferase reporter gene assay.Results:Starbase database analysis found that the standardized expression level of miR-886-5p in 370 liver cancer samples was lower than that in normal liver tissue samples (0.12±0.07 vs. 0.73±0.27, t=-15.71, P<0.001), and the expression level of miR-886-5p was positively correlated with the expression level of BAX ( r=0.152, P=0.003). qRT-PCR analysis showed that the expression level of miR-886-5p in HL-7702 cells was higher than that in Huh7 (4.57±0.06 vs. 1.61±0.40, t=32.48) and HepG2 (4.57±0.06 vs. 1.03±0.13, t=143.9), and the expression level of BAX in HL-7702 cells was higher than that in Huh7 (4.01±0.12 vs. 1.28±0.09, t=82.20) and HepG2 (4.01±0.12 vs. 1.30±0.11, t=80.76), the differences were statistically significant (all P<0.001). The proliferation, migration, and invasion abilities of Huh7 and HepG2 cells decreased after transfection with miR-886-5p mimics, while the expression levels of BAX at the mRNA and protein levels increased. However, after inhibiting the expression of miR-886-5p, the above indicators of cells were the opposite, and the dif-ferences were statistically significant (all P<0.05). The viability, EdU positivity rate, cell migration rate, and number of transmembrane cells in the miR-886-5p+ BAX group were lower than those in the BAX group, and the relative expression levels of miR-886-5p, BAX mRNA, and BAX protein were higher than those in the BAX group. However, the above indicators in the Sponge+ BAX group showed opposite trends, and all differences were statistically significant (all P<0.05). There was a targeted binding site between miR-886-5p and BAX. Conclusion:Both miR-886-5p and BAX are downregulated in liver cancer, and miR-886-5p inhibits the proliferation, migration, and invasion of liver cancer cells by targeting BAX.

Objective:To investigate the safety and efficacy of trans-breast endoscopic approach for lateral neck dissection (LND) in the treatment of thyroid cancer.Methods:A retrospective analysis was conducted on 80 patients who underwent trans-breast endoscopic LND at the Second Affiliated Hospital of Zhejiang University School of Medicine from Dec. 2023 to Aug. 2024. Demographic characteristics, operative duration, postoperative complications, and pathological results were analyzed, with data presented using descriptive statistics.Results:The cohort included 7 males and 73 females, with a mean age of (30.0 ± 6.7) years. The mean diameter of thyroid cancer lesion was (1.3 ± 0.7) cm. The average operative duration was (4.6 ± 1.5) hours, with a 0.0% (0/80) rate of conversion to open surgery. Intraoperative recurrent laryngeal nerve (RLN) "loss of signal" occurred in 6.3% (5/80) of cases. The mean hospital stay was (2.3 ± 1.0) days. Postoperative complications included transient hoarseness in 3.8% (3/80) and transient hypoparathyroidism in 10.0% (8/80), with no cases of permanent hoarseness or hypoparathyroidism. The mean total number of dissected lymph nodes was 47.8 ± 18.6, with 8.3 ± 6.2 metastatic lymph nodes. The incidence of postoperative lymphatic leakage was 3.8% (3/80), and no cases of incision infection were observed. The mean visual analog scale (VAS) score for pain was 4.2 ± 0.4. The mean follow-up duration was 10.3 months, at 3-month follow-up, the cosmetic satisfaction score was 4.5 ± 0.4.Conclusions:The trans-breast endoscopic approach for LND is safe and effective, with high patient cosmetic satisfaction. With appropriate patient selection, it can be considered a viable surgical option for lateral neck dissection in the treatment of thyroid cancer.

Objective:To investigate the safety and efficacy of trans-breast endoscopic approach for lateral neck dissection (LND) in the treatment of thyroid cancer.Methods:A retrospective analysis was conducted on 80 patients who underwent trans-breast endoscopic LND at the Second Affiliated Hospital of Zhejiang University School of Medicine from Dec. 2023 to Aug. 2024. Demographic characteristics, operative duration, postoperative complications, and pathological results were analyzed, with data presented using descriptive statistics.Results:The cohort included 7 males and 73 females, with a mean age of (30.0 ± 6.7) years. The mean diameter of thyroid cancer lesion was (1.3 ± 0.7) cm. The average operative duration was (4.6 ± 1.5) hours, with a 0.0% (0/80) rate of conversion to open surgery. Intraoperative recurrent laryngeal nerve (RLN) "loss of signal" occurred in 6.3% (5/80) of cases. The mean hospital stay was (2.3 ± 1.0) days. Postoperative complications included transient hoarseness in 3.8% (3/80) and transient hypoparathyroidism in 10.0% (8/80), with no cases of permanent hoarseness or hypoparathyroidism. The mean total number of dissected lymph nodes was 47.8 ± 18.6, with 8.3 ± 6.2 metastatic lymph nodes. The incidence of postoperative lymphatic leakage was 3.8% (3/80), and no cases of incision infection were observed. The mean visual analog scale (VAS) score for pain was 4.2 ± 0.4. The mean follow-up duration was 10.3 months, at 3-month follow-up, the cosmetic satisfaction score was 4.5 ± 0.4.Conclusions:The trans-breast endoscopic approach for LND is safe and effective, with high patient cosmetic satisfaction. With appropriate patient selection, it can be considered a viable surgical option for lateral neck dissection in the treatment of thyroid cancer.

Objective:To investigate the molecular mechanism of miR-886-5p targeting BCL-2-associated X protein (BAX) to inhibit the proliferation, migration, and invasion of liver cancer cells.Methods:mRNA expression data of HCC patients were obtained from the Starbase database, including 370 liver cancer samples and 50 normal liver tissue samples adjacent to the cancer. Analyze the expression of miR-886-5p in the previously obtained data and investigate the relationship between miR-886-5p and BAX in liver cancer samples. After transfection of the corresponding plasmids into Huh7 and HepG2 cells, the following groups were established. Analyze the interaction between miR-886-5p and BAX in vitro, detect the protein expression by Western blotting, and verify the targeting relationship between the two by dual luciferase reporter gene assay.Results:Starbase database analysis found that the standardized expression level of miR-886-5p in 370 liver cancer samples was lower than that in normal liver tissue samples (0.12±0.07 vs. 0.73±0.27, t=-15.71, P<0.001), and the expression level of miR-886-5p was positively correlated with the expression level of BAX ( r=0.152, P=0.003). qRT-PCR analysis showed that the expression level of miR-886-5p in HL-7702 cells was higher than that in Huh7 (4.57±0.06 vs. 1.61±0.40, t=32.48) and HepG2 (4.57±0.06 vs. 1.03±0.13, t=143.9), and the expression level of BAX in HL-7702 cells was higher than that in Huh7 (4.01±0.12 vs. 1.28±0.09, t=82.20) and HepG2 (4.01±0.12 vs. 1.30±0.11, t=80.76), the differences were statistically significant (all P<0.001). The proliferation, migration, and invasion abilities of Huh7 and HepG2 cells decreased after transfection with miR-886-5p mimics, while the expression levels of BAX at the mRNA and protein levels increased. However, after inhibiting the expression of miR-886-5p, the above indicators of cells were the opposite, and the dif-ferences were statistically significant (all P<0.05). The viability, EdU positivity rate, cell migration rate, and number of transmembrane cells in the miR-886-5p+ BAX group were lower than those in the BAX group, and the relative expression levels of miR-886-5p, BAX mRNA, and BAX protein were higher than those in the BAX group. However, the above indicators in the Sponge+ BAX group showed opposite trends, and all differences were statistically significant (all P<0.05). There was a targeted binding site between miR-886-5p and BAX. Conclusion:Both miR-886-5p and BAX are downregulated in liver cancer, and miR-886-5p inhibits the proliferation, migration, and invasion of liver cancer cells by targeting BAX.

Objective:To investigate the effect of circular RNA-Hsa-0101216 (Hsa-circ-0101216) targeting microRNA-142-3p (miR-142-3p) on the proliferation,cloning,migration and invasion of pancreatic cancer cells, and explore its molecular mechanism.Methods:The differentially expressed miRNAs in pancreatic cancer were analyzed and screened with GEO database and the circRNA-miRNA network was constructed. The expression of Hsa-circ-0101216 miRNA and miR-142-3p in 5 strains of pancreatic cancer cells (BxPC-3, PANC1, MIA PaCa-2, Capan-2, CFPAC-1) and normal pancreatic duct epithelial cells (HPNE) was detected by quantitative real time PCR. The PANC1 and Capan-2 pancreatic cancer cells were divided into the Hsa-circ-0101216 small interference RNA transfection group (si-circRNA group), the senseless negative sequence siRNA transfection group (si-NC group), and the normal blank control (NC group); the miR-142-3p overexpression lentiviral vector transfection group (miR-142-3p mimic group), the empty vector transfection group (miR-CON group), the co transfection of Hsa-circ-0101216 siRNA and miR-142-3p mimic group (si-Hsa circ0101216+mimic miR-142-3p group), co transfection of Hsa-circ-0101216 siRNA and miR-142-3p mimic negative control sequence group (si-Hsa-circ-010126+mimic miR-142-3p-NC group). The changes of cell proliferation, cloning, migration and invasion were detected by CCK-8, EdU proliferative staining, plate cloning, cell scratch and Transwell assay. Detect the expression of ERK1 and ERK1 and ERK2 protein expression was measured by Western blotting, and the targeting relationship between Hsa-cic-0101216 and miR-142-3p was verified by dual luciferase reporter gene method.Results:The expression of miR-142-3p was significantly down-regulated by GEO analysis, and the Hsa-circ-0101216-miR-142-3p regulatory network was successfully constructed. The mRNA expression levels of Hsa circ0101216 in BxPC-3, PANC1, MIA PaCa-2, Capan-2, and CFPAC-1 cells were 5.64±0.34, 5.93±0.40, 5.66±0.14, 5.63±0.33, and 5.70±0.50, respectively, which were significantly higher than those in HPNE cells (1.27±0.06); the expression levels of miR-142-3p were 1.43±0.12, 1.20±0.09, 1.60±0.04, 1.16±0.25, and 1.42±0.11, respectively, which were significantly lower than those of HPNE cells (4.69±0.22), and all the differences were statistically significant (all P value <0.001). Among them, the expression level of Hsa-circ-0101216 mRNA in PANC1 cells was the highest, and the expression level in Capan 2 cells was the lowest. Compared with the NC group and si-NC group, the si-circRNA group showed a significant decrease in the absorbance value at 450 nm at 24, 48, and 72 hours ( A450 value), EdU positivity rate, cell clone number, migration rate, and transmembrane cell number of PANC1 and Capan-2 cells; the A450 value at the above time points, EdU positivity rate, cell clone number, migration rate, and transmembrane cell number of PANC1 and Capan-2 cells in the miR-142-3p mimic group were significantly lower than those in the NC group and miR-CON group. The A450 value, EdU positivity rate, cell clone number, migration rate, transmembrane cell number, ERK1 and ERK2 protein expression levels of PANC1 and Capan-2 cells in the si-Hsa-circ-0101216+mimic-miR-142-3p group were significantly reduced compared to the si-Hsa-circ-0101216+mimic miR-142-3p-NC group, and the differences were statistically significant (all P<0.001).The luciferase activity of PANC1 and Capan-2 cells in co transfected with wild-type (WT) Hsa-circ-0101216 and miR-142-3p mimic groups was significantly lower than that of the miR-CON+WT group (0.92±0.11 vs 2.33±0.21, 0.89±0.08 vs 2.30±0.17), and the differences were statistically significant (all P value <0.001), but there was no statistically significant difference on luciferase activity of PANC1 and Capan-2 cells between the co transfected mutant (MUT) Hsa-circ-0101216 and miR-142-3p mimic groups and the miR-CON+MUT group. Conclusions:Hsa-circ-0101216 is overexpressed in pancreatic cancer cells, while miR-142-3p is poorly expressed; Hsa-circ-0101216 can promote the proliferation, cloning, migration and invasion of pancreatic cancer cells by targeting miR-142-3p.

Objective:To investigate the effect of circular RNA-Hsa-0101216 (Hsa-circ-0101216) targeting microRNA-142-3p (miR-142-3p) on the proliferation,cloning,migration and invasion of pancreatic cancer cells, and explore its molecular mechanism.Methods:The differentially expressed miRNAs in pancreatic cancer were analyzed and screened with GEO database and the circRNA-miRNA network was constructed. The expression of Hsa-circ-0101216 miRNA and miR-142-3p in 5 strains of pancreatic cancer cells (BxPC-3, PANC1, MIA PaCa-2, Capan-2, CFPAC-1) and normal pancreatic duct epithelial cells (HPNE) was detected by quantitative real time PCR. The PANC1 and Capan-2 pancreatic cancer cells were divided into the Hsa-circ-0101216 small interference RNA transfection group (si-circRNA group), the senseless negative sequence siRNA transfection group (si-NC group), and the normal blank control (NC group); the miR-142-3p overexpression lentiviral vector transfection group (miR-142-3p mimic group), the empty vector transfection group (miR-CON group), the co transfection of Hsa-circ-0101216 siRNA and miR-142-3p mimic group (si-Hsa circ0101216+mimic miR-142-3p group), co transfection of Hsa-circ-0101216 siRNA and miR-142-3p mimic negative control sequence group (si-Hsa-circ-010126+mimic miR-142-3p-NC group). The changes of cell proliferation, cloning, migration and invasion were detected by CCK-8, EdU proliferative staining, plate cloning, cell scratch and Transwell assay. Detect the expression of ERK1 and ERK1 and ERK2 protein expression was measured by Western blotting, and the targeting relationship between Hsa-cic-0101216 and miR-142-3p was verified by dual luciferase reporter gene method.Results:The expression of miR-142-3p was significantly down-regulated by GEO analysis, and the Hsa-circ-0101216-miR-142-3p regulatory network was successfully constructed. The mRNA expression levels of Hsa circ0101216 in BxPC-3, PANC1, MIA PaCa-2, Capan-2, and CFPAC-1 cells were 5.64±0.34, 5.93±0.40, 5.66±0.14, 5.63±0.33, and 5.70±0.50, respectively, which were significantly higher than those in HPNE cells (1.27±0.06); the expression levels of miR-142-3p were 1.43±0.12, 1.20±0.09, 1.60±0.04, 1.16±0.25, and 1.42±0.11, respectively, which were significantly lower than those of HPNE cells (4.69±0.22), and all the differences were statistically significant (all P value <0.001). Among them, the expression level of Hsa-circ-0101216 mRNA in PANC1 cells was the highest, and the expression level in Capan 2 cells was the lowest. Compared with the NC group and si-NC group, the si-circRNA group showed a significant decrease in the absorbance value at 450 nm at 24, 48, and 72 hours ( A450 value), EdU positivity rate, cell clone number, migration rate, and transmembrane cell number of PANC1 and Capan-2 cells; the A450 value at the above time points, EdU positivity rate, cell clone number, migration rate, and transmembrane cell number of PANC1 and Capan-2 cells in the miR-142-3p mimic group were significantly lower than those in the NC group and miR-CON group. The A450 value, EdU positivity rate, cell clone number, migration rate, transmembrane cell number, ERK1 and ERK2 protein expression levels of PANC1 and Capan-2 cells in the si-Hsa-circ-0101216+mimic-miR-142-3p group were significantly reduced compared to the si-Hsa-circ-0101216+mimic miR-142-3p-NC group, and the differences were statistically significant (all P<0.001).The luciferase activity of PANC1 and Capan-2 cells in co transfected with wild-type (WT) Hsa-circ-0101216 and miR-142-3p mimic groups was significantly lower than that of the miR-CON+WT group (0.92±0.11 vs 2.33±0.21, 0.89±0.08 vs 2.30±0.17), and the differences were statistically significant (all P value <0.001), but there was no statistically significant difference on luciferase activity of PANC1 and Capan-2 cells between the co transfected mutant (MUT) Hsa-circ-0101216 and miR-142-3p mimic groups and the miR-CON+MUT group. Conclusions:Hsa-circ-0101216 is overexpressed in pancreatic cancer cells, while miR-142-3p is poorly expressed; Hsa-circ-0101216 can promote the proliferation, cloning, migration and invasion of pancreatic cancer cells by targeting miR-142-3p.

BACKGROUND: Docetaxel is a commonly used anti-tumor drug in clinic, especially as the first-line drug for advanced non-small cell lung cancer (NSCLC). However, the molecular mechanism of docetaxel against NSCLC is still unclear. Increasing studies have shown that metabolic reprogramming of tumor cells plays an important role in tumorigenesis. The aim of this study was to investigate the effects of docetaxel on the metabolic pathway of NSCLC cells based on metabolomics analysis and biological means. METHODS: First, we performed CCK8 assay to analyze the effects of docetaxel on cell viability of NSCLC cells and also to screen the appropriate drug concentration. Then, the differential metabolites of docetaxel-treated and untreated NSCLC cells were analyzed by gas chromatography-mass spectrometry based metabolomics. Finally, the effects of docetaxel on the expression levels of key enzymes that regulate the relevant metabolic pathways were determined by Western blot. RESULTS: Docetaxel inhibited cell viability of A549 and H1299 cells in a concentration- and time-dependent manner. With the prolonged treatment time of docetaxel, the apoptotic sensitive protein poly (ADP-ribose) polymerase (PARP) was gradually activated to form a P89 fragment. Metabolomics analysis showed that eight metabolites were significantly changed in both A549 and H1299 cells following docetaxel treatment, which were mainly in the tricarboxylic acid (TCA) cycle pathway. Moreover, after docetaxel treatment, the protein expression levels of isocitrate dehydrogenases, the key regulators of the TCA cycle, were obviously decreased in both A549 and H1299 cells. CONCLUSIONS Our findings suggest that the effect of docetaxel-induced proliferation inhibition and apoptosis in NSCLC might be associated with down-regulation of isocitrate dehydrogenases and suppression of the TCA cycle pathway.
A549 Cells ; Apoptosis ; drug effects ; Carcinoma, Non-Small-Cell Lung ; pathology ; Docetaxel ; pharmacology ; Humans ; Lung Neoplasms ; pathology ; Metabolomics

Objective: To evaluate the predictive accuracy of fine needle aspiration (FNA) and BRAF V600E mutation in distinguishing papillary thyroid carcinoma and other thyroid nodules. Methods: This retrospective cohort study included 93 patients with papillary thyroid carcinoma who treated at Department of Thyroid Surgery, the Second Affiliated Hospital of Zhejiang University, College of Medicine from September 2016 to May 2018. There were 21 males and 72 females with age of (43.2±11.3) years (range: 19 to 67 years). All the patients got the examinations of FNA and BRAF V600E mutation by Amplification Refractory Mutation System, and subsequently underwent thyroid surgeries. The results of cytopathology, frozen section and pathology were collected and analyzed. The predictive accuracy of FNA cytology and BRAF V600E mutation was calculated. Results: In the 93 collected cases, 91 were diagnosed as papillary thyroid carcinoma postoperation, and the accurate predictive rate was 97.8%. Subgroup analysis was performed according to Bethesda System, the predictive rates were: unsatisfactory (Ⅰ) 6/6, benign (Ⅱ) 0/0, atypia of undetermined significance or follicular lesion of undetermined significance (Ⅲ) 16/17, follicular neoplasm or suspicious for follicular neoplasm (Ⅳ) 97.2% (35/36), suspicious for malignancy (Ⅴ) 100% (28/28), and malignant (Ⅵ) 6/6, respectively. Conclusion Thyroid nodules with BRAF V600E mutation can be strongly speculated as papillary thyroid carcinoma.

Objective To explore the technique and significance of intraoperative neuromonitoring (IONM) for scarless in the neck endoscopic thyroidectomy (SET) via breast approach.Methods From Apr.2015 to Oct.2015,101 consecutive patients undergoing SET with IONM were included.During the operation,patients received radical resection of the thyroid cancer by Wang's seven-step method.The lymph nodes in the central area were dissected and Wang's multi-functional separation forceps were implemented for recurrent laryngeal nerve (RLN) positioning,monitoring and protection.Also,time required for RLN positioning and exposure,postoperative transient and permanent RLN damage incidence were calculated to assess the feasibility of IONM under SET.Results Among 101 patients,130 RLNs in total were exposed.The average time required for RLN positioning under IONM was (3.26 ± 1.08)min,with round-nerve management time of (13.95 ± 4.58)min.Nerve signal change happened in 16.9％(22/130) patients.Positive predictive value was 13.6％ and negative predictive value was 100％.The overall accuracy rate was 85.4％.Conclusion IONM during SET is feasible,and can be helpful for the localization and functional protection of RLN and was useful to predict vocal cord paralysis.

Objective To investigate the immediate oral feedback after objective structure clinical examination (OSCE) for postgraduate year 1 & 2 surgery residents (PGY1 & 2).Methods 37 PGY1 and 38 PGY2 wereevaluated.The examination was composed of 6 stationsand limited to15 minutes per station.Each station was evaluated by centesimal system score.Immediate oral feedback was given in the last2 minutes.A questionnaire was given to each resident and examiner at the end of OSCE.All data analyses were conducted using SPSS version 22.0,repeated measures ANOVA and LSD test were used,and correlations were tested by the Pearson correlation test.Results The average scores for PGY1 & 2 were (68.97 ± 5.40) and (68.35 ± 5.00),the between-and inter-round differences in average score were not statistically significant.There was no significant correlation about theevaluation of the residents' performance during OSCE between the examiners and the residents.The necessity and effectiveness of immediate oral feedback were confirmed by both the examiners and the residents.Conclusions Immediate oral feedback isfeasible with limited impact on OSCE score,but the plan should be furtherrefined.Follow-up study isnecessary to identify the long-term effect on the clinical competency.