[ABSTRACT]OBJECTIVEThis study aims to explore the molecular mechanism and expression of Wnt/β-catenin signaling pathway and tumor marker CD44 in nasopharyngeal carcinoma cells after transfection withβ-Catenin when the Wnt/β-catenin signaling pathway was blocked.METHODSSP cells and CD44+SP cells isolated from the nasopharyngeal carcinoma cell line CNE-2 by flow cytometry were identified. Changes in the number and biological characteristics of CNE-2 and CD44+SP cells in vitro were investigated after the Wnt/β-catenin signaling pathway was blocked through siRNA.RESULTSSP cells accounted for 2.3% of nasopharyngeal carcinoma CNE-2 cells, andCD44+SP cells accounted for 36.5% of the SP cells. CD44+SP cells showed significantly higher in vitro proliferation and resistance to chemotherapy (P<0.05). After transfection withβ-Catenin siRNA, the proliferation, cloning efficiency, and tolerance to chemotherapeutic drugs of the cells were found statistical differences compared with those before transfection ofβ-Catenin siRNA. CONCLUSIONWnt/β-catenin signaling pathway abnormalities are closely related to the biological behavior of nasopharyngeal carcinomaCD44+SP cells. Interference of this pathway can change the characteristics of nasopharyngeal carcinoma stem cells.

OBJECTIVE: Discussion of expression and its significance of CD44 in SP cells of nasopnaryngeal carcinoma. METHOD: Flow cytometry was used to sort cultured CNE-2 cells of nasopharyngeal carcinoma for obtaining CD44-SP and CD44+SP cells. Biological differences of CNE-2, CNE-2 SP, CNE-2 NSP, CNE-2 CD44+SP and CNE-2 CD44-SP cells were statistically analyzed by experiments such as cell migration experiments, plate clone formation assay, cell cycle analysis and sensitivity tests to chemotherapeutics. RESULT: Two point 3 perent of SP cells were extracted from CNE-2 cells of nasopharyngeal carcinoma, among which 36.5% was CD44+SP cells. Abilities of proliferation, cell migration and plate clone of CD44+SP cells were significantly higher than other cells (P < 0.01), and its tolerance to chemotherapeutics was significantly higher too (P < 0.01). CONCLUSION The proportion of SP cells in nasopharyngeal carcinoma cells was small, but SP cells had strong activeness in the aspect of cell proliferation with a "seed" characteristic of tumor cells. As CD44+SP cells played an important role in proliferation and chemotherapy resistance of nasopharyngeal carcinoma, it indicated that CD44 can be one of the surface markers of SP cells of nasopharyngeal carcinoma.
Biomarkers, Tumor ; metabolism ; Carcinoma ; Cell Cycle ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Flow Cytometry ; Humans ; Hyaluronan Receptors ; metabolism ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; metabolism

Objective To investigate the image features of congenital pyriform sinus fistula (CPSF). Methods We retrospectively analyzed the clinical features and preoperative images of 80 patients with confirmed diagnosis of CPSF by surgical and pathological outcome in Guangdong general hospital from January 2007 to December 2014. At least one of the following imaging examinations were performed for all the patients, including Barium swallow X-ray (BSX), CT and MRI. Among them, 63 patients were examined with BSX, while 42 patients underwent plain and enhanced CT scans, wherein 40 of them were exanimated shortly after BSX. Thirty-two patients underwent plain and enhanced MRI scans. Patients were divided into two groups according to their age, young age group (≤14 years old) and older age group (>14 years old). Furthermore, they were also grouped based on inflammatory or quiescent stage clinically. The images of BSX, CT, and MRI from the patients were analyzed and the positive diagnostic rates (PDR) between groups were compared by using χ2 tests. Results For the patients examined with BSX, sinuses in 35 of 63 were depicted from pyriform and fistulas in 9 of 63 were depicted from the pyriform. The overall PDR of BSX was 74.6%(47/63),wherein 46.2%(12/26)in young age group , 94.6%(35/37)in older age group, 52.9%(9/17) in inflammatory stage group ,and 82.6%(38/46)in quiescent stage group. The inter-group differences were statistically significant (χ2 were 18.911 and 5.766,both P<0.05). The PDR of CPSF with CT was 85.7%(36/42), MRI was 84.4%(27/32), BSX+CT was 87.5%(35/40). The courses of fistula or sinus were showed on CT and MRI. The presence of air bubbles at the inferomedial edge of cricothyroid joints or around the upper lobe of the thyroid gland, the changes of the morphology of thyroid grand as well as the inflammatory change along the fistula region were detected much clearly on CT and MRI. There was no statistical difference between CT and MRI groups(P>0.05).Conclusions BSX could be a screening method for suspected cases of CPSF in quiescent stage. However, the PDR could be affected by many factors (age and inflammation). CT and MRI could provide valuable information for diagnosis. An examination combined BSX and CT is preferred to improve the positive detective rate of CPSF.

Objective To evaluate the effectiveness of endoscopic CO2 laser cauterization (ECLC) as a definitive treatment of congenital pyriform sinus fistula (CPSF).Methods Eleven patients with CPSF underwent ECLC between January 2011 to March 2013 at Guangdong General Hospital.Of the 11 patients aging from 20 to 672 months (median:60 months),there were 4 males and 7 females; 10 lesions located in the left necks and 1 located in the right; 6 untreated previously and 5 recurrent; 10 sinus (with internal opening) and 1 fistula.The presentations included reduplicative neck swelling,pain,or a fistulous opening with purulent discharge at the anterior neck region.Preoperative examinations included barium esophagogram,CT,MRI and so on.Six patients had at least received one time incision and drainage procedure previously.All patients had been treated with antibiotics in acute infection period.After inflammation subsided,the openings of pyriform sinus fistula were confirmed by esophagoscopy and then ECLC on internal opening was routinely performed.Esophagoscopy was carried out again by 3 months later in every patient,the same technique would be performed immediately if the internal opening was not completely closed.Results The existence of an orifice in the pyriform fossa was identified by esophagoscopy in 11 patients.In 9 patients,the treatment was successful and the internal opening completely closed after the first ECLC.However,the other 2 patients received the second cauterization 3 months later because of the incomplete close of the internal opening.The average number of treatments was 1.2 times.No complications such as dysphagia,hoarseness occurred with the endoscopic procedure.Both the patients and their families were satisfied with the cervical appearance.All the patients had an uneventful recovery and remained no symptom from 11 to 35 months (median:24 months).Conclusion The endoscopic CO2 laser cauterization is safe,effective,repeatable and minimally invasive,which can be suggested as first-line treatment for congenital pyriform sinus fistula.

Objective:To investigate the clinical value of ultrasound in evaluating pharyngeal dysphagia after stroke.Methods:Thirty stroke survivors with dysphagia formed the patient group, and 30 healthy persons formed the healthy group. All received pharyngeal ultrasound examinations by the same sonographer. The maximum distance from the hyoid bone to the mandible, the minimum distance, and the required time were measured or calculated. The hyoid bone′s motion, distance shortening rate, geniohyoid muscle activity, activity time, and activity speed were calculated. The differences in each value were compared, and the values were correlated with the videofluoroscopic swallowing study (VFSS) score using Spearman rank correlation. Receiver operating characteristics curves (ROC curves) were used to evaluate the value of hyoid bone and geniohyoid muscle activity for evaluating the severity of pharyngeal swallowing disorders after a stroke.Results:The average range of activity, activity time, and activity speed of the geniohyoid muscle in the patient group were all significantly different from the healthy group′s averages. The hyoid bone′s range of activity, activity time, activity speed, and the rate of shortening of its distance from the mandible were also significantly different, on average. The patients′ VFSS grades correlated strongly with the activity of the geniohyoid muscle, moderately with the geniohyoid muscle′s speed, and weakly with the duration of hyoid bone activity.Conclusion:The speed of geniohyoid muscle activity and hyoid bone activity time can be used to evaluate the severity of pharyngeal swallowing dysfunction after a stroke. Geniohyoid muscle activity is the best indicator.

Abstract: To investigate the changes of cerebral blood flow(CBF) in patients with type 2 diabetes mellitus(T2 DM) and its correlation with cognitive function and olfactory impairment. Methods: Cognitive function assessment and smell identification test were performed on 83 patients with T2 DM and 62 healthy controls(HC). Three-dimensional pseudo-continuous arterial spin labeling(3 D-pcASL) head images were collected from the two groups. CBF values of the cerebral cortex were compared between the patients and HC after the postprocessing. Correlations between the CBF values and cognitive function assessment and between the CBF values and smell identification test scores were analyzed as well. Results: Compared to the HC, Chinese smell identification test(CSIT), montreal cognitive assessment(MoCA), digit span test(DST), verbal fluency test(VFT) scores were lower in T2 DM patients(P<0.05).The CBF of the bilateral middle frontal gyrus in T2 DM patients was higher than that in HC group(P<0.001). The CBF of the bilateral gyrus rectus and olfactory cortex in T2 DM patients was lower than that in HC group(P<0.001). Conclusion The cognitive and olfactory function of patients with T2 DM decreased. Patients with T2 DM have abnormal perfusion in the bilateral middle frontal gyrus, gyrus rectus and olfactory cortex, revealing that CBF changes in these brain regions may be one of the causes for cognitive impairment and olfactory dysfunction in T2 DM.

Objective: To investigate the anatomic tract of congenital pyriform sinus fistula (CPSF). Methods: A total of 90 patients with CPSF undergoing open surgery between August, 2007 and March, 2017 at the Department of Guangdong General Hospital were retrospectively analyzed. Results: The tracts of all the fistulas actually walked far different from those of theoretical ones. A whole fistula may be divided into 4 segments according to adjacent anatomy of CPSF. The posterior inner segment to the thyroid cartilage was initial part of the fistula. It originated from the apex of pyriform sinus, then piercing out of the inferior constrictor of pharynx inferiorly near the inferior cornu of the thyroid cartilage (ICTC), and descended between the lateral branch of the superior laryngeal nerve and the recurrent laryngeal nerve. The ICTC segment was the second part of the fistula, firstly piercing out of the inferior constrictor of pharynx and/or cricothyroid muscle, and then entering into the upper pole of thyroid. The relationship between fistula and ICTC could be divided into three types: type A (medial inferior to ICTC) accounting for 42.2% (38/90); type B (penetrate ICTC) for 3.3% (3/90); and type C (lateral inferior to ICTC) for 54.5% (49/90). The internal segment in thyroid gland was the third part of fistula, walking into the thyroid gland and terminating at its upper pole (92.2%, 83/90) or deep cervical fascia near the upper pole of thyroid (7.8%, 7/90). The lateral inferior segment to thyroid gland was the last part of the fisula, most of which are iatrogenic pseudo fistula, and started from the lateral margin of thyroid gland. Conclusions CPSF has a complicated pathway. Recognition of the tract and adjacent anatomy of CPSF will facilitate the dissection and resection of CPSF in open surgery.