Objective To investigate the safety and feasibility of modified gasless trans-subclavian approach endoscopic total thyroidectomy for papillary thyroid carcinoma(PTC).Methods We retrospectively analyzed the clinical data of consecutive 82 patients with PTC who underwent the modified gasless trans-subclavian approach endoscopic total thyroidectomy in the Department of Head and Neck Surgery,Sir Run Run Shaw Hospital,Affiliated with the Zhejiang University School of Medicine,from March 2022 to June 2023.Results All operations were successfully completed under endoscopy approach without transfer to open surgery.The maximum diameter of PTC was(6.6±5.4)mm.The operation duration was(156.5±32.7)min,and the number of lymph nodes in the central compartment was(12.3±6.4).The postoperative hospital stay was(3.3±1.2)d.Postoperative complications included two cases of transient hypoparathyroidism and six cases of transient recurrent laryngeal nerve injury.All the patients recovered in one month.One case of postoperative bleeding was observed.No permanent laryngeal nerve injury,permanent hypoparathyroidism or incision infection occurred.Conclusion The gasless trans-subclavian approach endoscopic total thyroidectomy is a feasible and safety approach in selected PTC patients.The lymph nodes in the central compartment are thoroughly dissected.This approach has obvious advantages in functional protection of the anterior cervical region,and has clinical application value.

AIM:To explore the possible mechanism of Yiqi-Huoxue formula(YQHXF)in treating cerebral ischemia-reperfusion injury.METHODS:Male SD rats were randomly divided into six groups,namely,the sham,mod-el,nimodipine,and low-,middle-and high-dose YQHXF groups.The middle cerebral artery occlusion/reperfusion(MCAO/R)model was established in all groups except the sham group.After successful modeling,the YQHXF low-,me-dium-,and high-dose groups were given 3.8,7.5,and 15 g?kg-1?d-1 of YQHXF,respectively,by gavage,while the ni-modipine group was given 12 mg?kg-1?d-1 of nimodipine tablets by gavage.The sham and model groups were given 10 mL?kg-1?d-1 of distilled water by gavage.After 14 days of drug intervention,the rats were euthanized and the neurological func-tion was evaluated.The infarct volume was assessed using 2,3,5-triphenyltetrazolium chloride(TTC)staining and brain histopathological changes were determined by hematoxylin-eosin(HE)staining.Transmission electron microscopy was used to investigate changes in autophagosomes,with immunofluorescence used to assess expression of microtubule-associ-ated protein 1 light chain 3(LC3)protein in the cerebral cortex,Western blot was used to measure protein levels of p-PI3K,PI3K,p-Akt,Akt,p-mTOR,mTOR,LC3B,p62,beclin-1,and Atg5,and RT-qPCR was used to determined LC3 and P62 mRNA expression.RESULTS:Compared with the sham group,the neural function scores of rats in the model group rats were significantly increased,and TTC staining revealed large areas of white cerebral infarction.There was severe pathological damage to the cerebral tissue in the ischemic cortical area,and large numbers of autophagosomes were seen inside the cells.Immunofluorescence staining showed significant numbers of LC3B-positive cells(P<0.01).Protein expression of beclin-1,Atg5,and LC3-Ⅱ/LC3-Ⅰ was significantly upregulated(P<0.01),while that of p62 was markedly downregulated(P<0.01).The expression of p-PI3K/PI3K,p-Akt/Akt,and p-mTOR/mTOR proteins was also significantly reduced(P<0.01).In addition,the mRNA expression of LC3 was significantly upregulated(P<0.01),with downregulation of P62 mRNA levels(P<0.01).Compared with the model group,both the YQHXF medium-and high-dose groups showed upregulated LC3-Ⅱ/LC3-Ⅰ values after 12 h of reperfusion(P<0.01),followed by downregulation of the ratios(P<0.05)after 3,7,and 14 days of reperfusion.Furthermore,after 14 days of reperfusion,compared with the model group,the middle-and high-dose YQHXF groups and the nimodipine group showed reduced neurological function scores(P<0.01),reduced cerebral infarction volumes(P<0.01),improvements in the pathological damage to cortical tis-sue,and reduced autophagosome formation to varying degrees.At the same time,the number of LC3B-positive cells was reduced(P<0.01).Protein expression of beclin-1,Atg5,and LC3-Ⅱ/LC3-Ⅰ was significantly downregulated,while that of p62 was upregulated(P<0.01).The mRNA expression of LC3 and p62 was consistent with the protein levels(P<0.01).In addition,the expression of p-PI3K/PI3K,p-Akt/Akt,and p-mTOR/mTOR proteins was upregulated(P<0.01).CONCLUSION:YQHXF can dynamically regulate autophagy in ischemic brain tissue,with inhibition of excess autophagy by activation of the PI3K/Akt/mTOR signaling pathway,thus reducing the infarct volume,alleviating brain dam-age,and promoting the recovery of neurological function.

Objective:To analyze the survival time, prognostic factors and the value of postoperative thoracic radiotherapy in resected small cell lung cancer (SCLC) patients.Methods:Clinic opathological data of SCLC patients who received surgical treatment in Cancer Hospital & General Hospital of Ningxia Medical University from April 2014 to September 2021 were enrolled in this retrospective study. All patients were subject to follow-up. The survival time of SCLC patients was evaluated by Kaplan-Meier method. Univariate and multivariate analyses of prognostic factors were performed by Cox proportional hazard model.Results:A total of 64 patients with SCLC were enrolled in the study. The 5-year overall survival (OS) rate was 43.5%. Univariate analysis showed that TNM staging ( P=0.027), postoperative neutrophil-lymphocyte ratio (NLR) ( P=0.039) and adjuvant thoracic radiotherapy ( P=0.041) were the prognostic factors. Multivariate analysis showed that TNM staging ( P=0.038) and adjuvant thoracic radiotherapy ( P=0.022) were the prognostic factors in patients with SCLC. The 5-year OS rates of patients with and without adjuvant thoracic radiotherapy were 71.6% and 35.4% ( P=0.028), respectively. There was a statistically significant difference in the 5-year OS rates between pathological stage N 2 SCLC patients with or without adjuvant thoracic radiotherapy (75.0% vs. 0%, P=0.030). Conclusions:TNM staging and postoperative adjuvant thoracic radiotherapy are prognostic factors in patients with SCLC undergoing surgical treatment. Pathological stage N 2 SCLC patients can benefit from adjuvant thoracic radiotherapy.

OBJECTIVE: To investigate the effect of Zuogui Jiangtang Jieyu Decoction (ZJJ) on Shh signaling and self-renewal of neural stem cells in the hippocampal dentate gyrus of diabetic rats with depression. METHODS: Diabetic rat models with depression were randomly divided into model group, positive drug (metformin + fluoxetine) group, and low-, medium-, and high-dose ZJJ groups (n=16), with normal SD rats as the control group. The positive drugs and ZJJ were administered by gavage, and the rats in the control and model groups were given distilled water. After the treatment, blood glucose level was detected using test strips, and behavioral changes of the rats were assessed by forced swimming test and water maze test. ELISA was used to examine the serum level of leptin; The expressions of nestin and Brdu proteins in the dentate gyrus of the rats were detected using immunofluorescence assay, and the expressions of self-renewal marker proteins and Shh signaling proteins were detected using Western blotting. RESULTS: The diabetic rats with depression showed significantly increased levels of blood glucose and leptin (P < 0.01) and prolonged immobility time in forced swimming test (P < 0.01) and increased stage climbing time with reduced stage seeking time and stage crossings in water maze test (P < 0.01). The expressions of nestin and Brdu in the dentate gyrus, the expressions of cyclin D1, SOX2, Shh, Ptch1, Smo in the hippocampus and the nuclear expression of Gli-1 were decreased (P < 0.01) while hippocampal Gli-3 expression was increased significantly (P < 0.01) in the rat models. Treatment of rat models with high-dose ZJJ significantly reduced the blood glucose (P < 0.01) and leptin level (P < 0.05) and improved their performance in behavioral tests (P < 0.01). The treatment also obviously increased the expressions of nestin, Brdu, cyclin D1, SOX2, Shh, Ptch1, and Smo and the nuclear expression of Gli-1 in the dentate gyrus (P < 0.01) and reduced hippocampal expression of Gli-3 (P < 0.05) in the rat models. CONCLUSION ZJJ can significantly improve the self-renewal ability of neural stem cells and activate Shh signaling in dentate gyrus of diabetic rats with depression.
Animals ; Rats ; Blood Glucose ; Bromodeoxyuridine ; Cell Self Renewal ; Cyclin D1 ; Dentate Gyrus ; Depression ; Diabetes Mellitus, Experimental ; Hippocampus ; Leptin ; Nestin ; Rats, Sprague-Dawley

Objective:To study the safety and feasibility of gasless transoral endoscopic thyroidectomy though vestibular approach using self-retaining retractor for papillary thyroid carcinoma.Methods:The clinical data of 39 papillary thyroid carcinoma patients undergoing gasless transoral endoscopic thyroidectomy were collected at Department of Head and Neck Surgery, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine from Nov 2020 to Jun 2021.Results:All cases successfully underwent laparoscopic surgery without conversion to open surgery. The mean duration of operation was (142±35) min, and the postoperative mean hospital stay was (4.1±0.8) days. The mean maximum diameter of the tumor was (8.5±4.5) mm, and the mean number of lymph node harvest of by central compartment dissection was 7.7±5.9. Postoperative complications were transient hypoparathyroidism in 2 cases but recovered in 1 month. Scalp hydrop in 1 patient,fading subsequently. Transient sensory change around the lower lip in 3 cases, which recovered in 6 months. No patient suffered from recurrent laryngeal nerve palsy or hematoma, no permanent hypoparathyroidism occurred, nor of the postoperative bleeding .Conclusion:The gasless transoral endoscopic thyroidectomy viaoral vestibular approach is a feasible approach in selected papillary thyroid carcinoma patients.

Objective:To investigate the expression of microRNA (miR)-206 in chronic obstructive pulmonary disease (COPD) and its effect on the proliferation of human airway smooth muscle cells (HASMCs) and to explore its mechanism.Methods:Lung tissue samples of 15 patients with COPD (COPD group) who underwent lung volume reduction surgery in the General Hospital of Ningxia Medical University from September 2017 to September 2018 and of 15 patients with benign lung tumors without a history of COPD were collected. Microarray technology was used to analyze the miR and RNA omics in lung tissues of 4 COPD patients and normal controls, and reverse transcriptase polymerase chain reaction(RT-PCR) was used to verify the results. Bioinformatics and double luciferase gene reporting assay were used to detect the target genes of miR-206 in HASMCs. The miR-206 mimic/inhibitor was transfected into HASMCs by liposome transfection technology, and the expression level of miR-206 was detected by RT-PCR. Methyl thiazolyl tetrazolium (MTT), flow cytometry and apoptosis assay were used to detect the effects of miR-206 on the proliferation, cell cycle and apoptosis of HASMCs. The expression of PTEN, cell cycle and apoptotic protein in HASMCs was detected by Western blot.Results:The results of miR and mRNA omics analysis showed that the expressions of miR-206, miR-3187-5p and miR-124 in COPD group were significantly up-regulated (0.09 ± 0.01 vs. 2.17 ± 0.57, 0.60 ± 0.04 vs. 1.32 ± 0.15, 0.22 ± 0.08 vs. 1.09 ± 0.23) ( P<0.05), while the expressions of miR-574 and miR-337-3p decreased significantly (0.79 ± 0.03 vs. 0.15 ± 0.02, 0.95 ± 0.02 vs. 0.17 ± 0.01) ( P<0.05). RT-PCR was used to detect the expression of these five miRNAs in 15 COPD lung tissues, and the results showed that their expression was consistent with that in microarray. The prediction results of miRNA target genes showed that miR-206 could directly inhibit the expression of PTEN. RT-PCR results showed that the expression of miR-206 in miR-206 transfected HASMCs was significantly higher than that in miR-NC transfected group(7.44 ± 0.51 vs. 4.02 ± 0.19), and miR-206 inhibitor could significantly inhibit the expression of miR-206 in cells (1.86 ± 0.32), the difference was statistically significant ( P<0.05); MTT and apoptosis experiments showed that miR-206 mimcs could significantly promote the proliferation rate of cells compared with normal HASMCs or miR-NC transfected cells (0.62 ± 0.14 or 0.57 ± 0.09 vs. 0.83 ± 0.05), inhibit cell apoptosis (9.13 ± 1.71 or 10.02 ± 1.15 vs. 3.06 ± 0.82), the differences were statistically significant ( P<0.05), while miR-206 inhibitor could significantly inhibit cell proliferation and promote cell apoptosis ( P<0.05) The results of cell cycle distribution showed that compared with HASMCs group, the proportion of cells in S phase and G2/M phase in miR-206 mimcs group increased significantly ( P<0.05), while the proportion of cells in S phase and G2/M phase in miR-206 inhibitor group decreased significantly ( P<0.05), and there was no significant difference in miR-NC group ( P>0.05). The results of Western blot showed that compared with normal HASMCs or miR-NC transfected cells, miR-206 mimcs could significantly upregulate the expression of cyclin D1 (0.43 ± 0.07 or 0.41 ± 0.02 vs. 0.63 ± 0.17), and cyclin B1 (0.47 ± 0.13 or 0.50 ± 0.09 vs. 0.79 ± 0.31), and inhibit the expression of PTEN (0.34 ± 0.10 or 0.29 ± 0.05 vs. 0.14 ± 0.02), cyclin p21 (0.34 ± 0.03 or 0.30 ± 0.05 vs. 0.11 ± 0.02), and apoptosis related protein caspase-3 (0.29 ± 0.03 or 0.31 ± 0.05 vs. 0.15 ± 0.03), the differences were statistically significant ( P<0.05). miR-206 inhibitor could significantly inhibit the expression of cyclin D1 and cyclin B1, and promote the expression of PTEN, cyclin p21 and caspase-3 ( P<0.05). Conclusions:In COPD patients, miR-206 could targeted inhibit the expression of PTEN protein in airway smooth muscle cells and regulate the progress of cell cycle, so as to up regulate the proliferation of cells and inhibit their apoptosis.

Objective To screen out the biomarkers with diagnostic value in lung cancer by biochip technology. Methods We screened four pairs of differentially-expressed circRNAs in lung cancer by circRNA expression profiling chip, and then verified the screened differential circRNA hsa_circ_0044569 by qRT-PCR, and collected clinical case information of patients at the same time. The independent sample t test and ROC curve were used to analyze the relation between the clinical data of lung cancer patients and the expression of circRNA hsa_circ_0044569 on clinical samples. Results The expression level of hsa_circ_0044569 in lung cancer tissues was higher than that of its paired adjacent tissues. The cutoff value of hsa_circ_0044569 for the diagnosis of lung cancer was 9.62, AUC was 0.758, P < 0.05, sensitivity was 0.712, and specificity was 0.712. The expression of hsa_circ_0044569 was significantly related with the pathological type of lung cancer patients (P < 0.05). Conclusion hsa_circ_0044569 is highly expressed in lung cancer and related to the pathological type, suggesting that it may become a potential biomarker in the early diagnosis of lung cancer.

To investigate the role of cardiac magnetic resonance imaging (CMRI) in evaluating pulmonary hemodynamics and right ventricular function in patients with chronic obstructive pulmonary disease (COPD) and pulmonary hypertension (PAH); and the relationship between CMRI parameters and pulmonary function parameters, blood gas analysis parameters and 6-minute walk test (6MWT) parameters in patients with COPD complicated with PAH. Methods Thirty-seven patients were diagnosed with COPD in the department of respiratory and critical care discipline of Ningxia Medical University General Hospital from October 2013 to October 2016, who underwent transthoracic echocardiography (TTE) to measure pulmonary arterial systolic pressure (PASP), and were divided into COPD group and COPD+PAH group according to whether there was PAH [PASP > 40 mmHg (1 mmHg = 0.133 kPa) was defined as PAH]. All patients completed pulmonary function tests [1 second forced expiratory volume to forced vital capacity ratio (FEV1/FVC), FEV1 predicted value (FEV1pred)], blood gas analysis [arterial blood oxygen partial pressure (PaO2), arterial blood carbon dioxide partial pressure (PaCO2)], CMRI examination [relative dilatation of the main pulmonary artery (mPAD), mean pulmonary artery pressure (mPAP), left ventricular ejection fraction (LVEF), right ventricular ejection fraction (RVEF), right ventricular end-diastolic myocardial mass (RVMED), right ventricular end-systolic myocardial mass (RVMES)], and 6MWD [6-minute walk distance (6MWD)] within 1 week. The obtained clinical parameters had been compared between the groups, and correlation was analyzed. Results Among the 37 patients with COPD, 16 patients were complicated with PAH. There were no significant differences in FEV1/FVC, FEV1pred, PaO2, PaCO2 and other baseline indicators between the two groups. In the COPD group, TTE obtained PASP of 2 patients were normal (PSAP < 40 mmHg), while CMRI measured mPAP were higher than the normal limit (> 25 mmHg). Compared with the COPD group, mPAD, RVEF and 6MWD were significantly decreased in the COPD+PAH group [mPAD: (25.64±5.01)% vs. (44.00±22.52)%, RVEF: 0.525±0.054 vs. 0.592±0.071, 6MWD (m): 319.3±116.5 vs. 408.2±38.0, all P < 0.01], mPAP, RVMED and RVMES were significantly increased [mPAP (mmHg): 28.89±3.16 vs. 20.18±2.43, RVMED (g): 57.19±15.46 vs. 40.71±15.44, RVMES (g): 45.99±11.16 vs. 33.71±13.39, all P < 0.01], and there was no significant differences in LVEF (0.663±0.082 vs. 0.699±0.075, P > 0.05). Correlation analysis showed that mPAD was positively correlated with FEV1/FVC and FEV1pred (r1 = 0.538, P1 = 0.021; r2 = 0.448, P2 = 0.049);RVMED was negatively correlated with PaO2 (r = -0.581, P = 0.015), and positively correlated with PaCO2 (r = 0.592, P = 0.014); 6MWD was positively correlated with RVEF (r = 0.485, P = 0.041), and had no correlation with LVEF (r = 0.271, P = 0.104). Conclusions Compared with COPD patients, changes in pulmonary hemodynamics and right ventricular function in COPD patients with PAH are related to the severity of airflow limitation. CMRI can early monitor pulmonary hemodynamics and right heart function changes in patients with COPD. Once PAH appears, pulmonary hemodynamics, right heart function and exercise tolerance have changed.

Objective To explore the application value of single-port inflatable mediastinoscopy combined with laparoscopy in the radical resection of esophageal cancer.Methods The retrospective descriptive study was conducted.The clinicopathological data of 27 patients who underwent single-port inflatable mediastinoscopic and laparoscopic radical resection of esophageal cancer in the General Hospital of Ningxia Medical University between September 2016 and April 2018 were collected.The surgical operators were divided into neck operation group and abdomen operation group.A "Y" tube was used to inflate the abdomen and mediastinum simultaneously with CO2,and the gas pressure was 12-16 mmHg (1 mmHg =0.133 kPa).Bilateral exchange free and join forces with the esophagus and xiphoid process operating small incision,the severed esophagus cardia;residual stomach was made into a 3-5 cm tubular stomach and was sutured at the top point;at the same time,esophagus was brought up from the neck,with a pouch suture between upper esophageal and stapling head;the tubular stomach through mediastinum-esophagus bed was pulled to the left neck and then gastroesophageal anastomosis manually or instrument was performed.Observation indicators:(1) surgical and postoperative recovery;(2) follow-up and survival situations.Follow-up using outpatient examination and telephone interview was performed to detect postoperative survival up to May 2018.The measurement data with normal distribution were represented as x-±s.The measurement data with skewed distribution were described as M (range).Results (1) Surgical and postoperative recovery:all the 27 patients underwent successful single-port inflatable mediastinoscopic and laparoscopic radical resection of esophageal cancer,with complete tumor resection and without conversion to open surgery.There was no arrhythmia or myocardial ischemia through intraoperative electrocardiography.Among 27 patients,5 had intraoperative rupture of the pleura and 3 stopped intermittently inflation with CO2 due to obvious hemodynamic changes.The operation time and volume of intraoperative blood loss were (121±21)minutes and (100± 30)mL.Twenty-seven patients had no thoracic incision,obviously decreased postoperative pain and out-of-bed activity at day 1 postoperatively.The volume of postoperative mediastinal drainage was (40± 10)mL.The mediastinal drainage-tube was removed at 1 week after regular food intake.Of 27 patients,5 with pleural effusion were cured by puncture drainage;2 were complicated with anastomotic leakage,1 of them with a small amount of subcutaneous gas under neck incision at 12 days postoperatively was cured spontaneously through oral food intake,without special treatment,and the other had a small amount of subcutaneous gas under neck incision after solid food intake at 1 month postoperatively and then was cured after 1-week fluid food intake;1 with anastomotic stenosis was improved after dilation treatment.The squamous cell carcinoma was confirmed by postoperative pathological examination,without cancer cell infiltration in the upper and lower margins.The numbers of mediastinal lymph node dissected,abdominal lymph nodes dissected and positive lymph node,postoperative pathological staging and duration of hospital stay were respectively 9.5±2.2,8.2±2.5,1 (range,0-12),T1-3N0-1M0 and 13 days (range,11-21 days).(2) Follow-up and survival situations:27 patients were followed up for 1-20 months,with a median time of 10 months.During the follow-up,there was no recurrence or metastasis and death.Conclusion The single-port inflatable mediastinoscopy combined with laparoscopy in the radical resection of esophageal cancer is safe and effective,and it is especially suitable for patients with partial respiratory failure and closed thoracic cavity.

BACKGROUND:Primary human lung epithelial cel s are difficult to be isolated and cultured in vitro, which is characterized as limited sources, low cel viability, slow proliferation capacity, and lacking of differentiation capability.
OBJECTIVE:To establish an air-liquid interface model of lung epithelium by in vitro proliferation of human bronchiolar epithelial cel s, which is used for research on function of lung epithelial cel s.
METHODS:Primary human bronchiolar epithelial cel s were isolated using Pronase and DNase I combined digestive methods, and then proliferated using medium containing ROCK kinase inhibitor. The proliferated cel s were used for establishment of the air-liquid interface epithelium model. Cel differentiation was identified using scanning electron microscope, phase contrast microscope and immunofluorescent staining.
RESULTS AND CONCLUSION:The primary human bronchiolar epithelial cel s could be expanded successful y using medium containing ROCK kinase inhibitor, and the basal cel marker Cytokeratin14 was preferential y expressed in the proliferated cel population, indicating that these basal cel s might be the main subpopulation of human lung epithelial stem cel s. Subsequently, the proliferated cel s under the air-liquid interface could differentiate into ciliated cel s and non-ciliated column cel s. The results suggest that the proliferation and differentiation of human bronchiolar epithelial cel s were maintained in the presence of ROCK kinase inhibitor, and the air-liquid interface could promote the differentiation of human bronchiolar epithelial cel s.