Objective To establish radiation?resistant lung carcinoma cell lines, and to investigate the changes in morphology, apoptosis, invasive migration, and epithelial?mesenchymal transition ( EMT) in cells. Methods The radiation?resistant lung carcinoma cell lines were obtained by exposure of lung carcinoma cell lines, A549 and H1299, to radiation with a low dose in fractions, a sublethal dose, or a gradually increasing dose. The morphological changes in cells, radiosensitivity, survival rates after exposure, apoptosis rates, changes in invasive migration, and expression of EMT marker proteins were evaluated using microscopy, colony formation assay, CCK?8 assay, flow cytometry, transwell migration assay, and Western blot, respectively. Results Radiation with a gradually increasing dose successfully induced the radiation?resistant cell lines, A549R and H1299R. The morphological study showed that the morphology of radiation?resistant cells was converted to the morphology of mesenchymal cells. Compared with A549 and H1299 cells, the values of D0 , Dq , and SF2 were significantly increased in A549R ( P=0.017,P=0.001,P=0.000) and H1299R (P=0.033,P=0.000,P=0.008) cells, respectively;the values of α and α/β were significantly reduced in A549R (P=0.018;P=0.007) and H1299R (P=0.001;P=0.009) cells, respectively. The survival rates in A549R and H1299R cells after exposure to radiation with various doses were significantly higher than those in the control groups (all P<0.05). After exposure, the apoptosis rates were significantly reduced in A549R and H1299R cells ( P=0.02,P=0.01);the invasion and migration rates were significantly increased in A549R (P=0.000;P=0.001) and H1299R (P=0.001,P=0.002) cells;the expression of E?cadherin was significantly down?regulated in A549R and H1299R cells (P=0.00,P=0.01), while the expression of vimentin was significantly elevated in A549R and H1299R cells ( P= 0. 02, P= 0. 01 ) . Conclusions The radiation?resistant lung carcinoma cell lines are successfully established. Both cell lines show enhanced invasion and migration, which may be associated with EMT.

Objective To study the molecular epidemiology of C.albicans isolates in infectious disease patients and to explore biofilm phenotypic characterization responsible for biofilm formation in clinical strains.Methods A total of 104 hospital-acquired C.alibcans clinical isolates collected from sterile sites and mucosal lesions of 92 infectious disease patients ( viral hepatitis, tuberculosis and AIDS) in Shanghai Public Health Clinical Center were analyzed.MLST analysis was performed to identify their phylogenetic status.The capability of biofilm formation was measured by [2,3-bis-(2-methoxy-4-nitro-5-sulphenyl)-2H-tetrazolium-5-carboxanilide] XTT assay.The results were compared using Kruskal-Wallis test.Results MLST analysis identified 63 DSTs with a decentralized phylogeny among 104 C.albicans isolates, of which 41 DSTs (65.1%) had not been reported in the online MLST database.The Single Locus Sequence Query from the C.albicans database identified new alleles.MEGA6 analysis of the MLST data assigned the 104 isolates within 14 of the 18 known clades; among them the clade 1 contained the greatest proportion of isolates (26.9%).Of the 43 novel DSTs isolates, 37 ( 86.0%) clustered within 11 of the 18 known clades.16 high biofilm formers were found from a total of 104 clinical isolates.The biofilm formation capabilities differed in strains isolated from different anatomical sites (H =18.23,P=0.0326).Biofilm formation by blood-originated isolates was lower than that of catheter-originated isolates ( Z=-72.20,P<0.001).Genotypes also affected the biofilm formation capability of the C.albicans isolates (H=10.01,P=0.0185).Conclusions A high level of diversity within C.albicans isolates.Microevlution clearly influences C.albicans genetic alterations upon environmental selection.The site of isolation and genotype associates with the biofilm formation capability.

Objective To understand the distribution and drug resistance of pathogens from patients with anastomotic fistula infection after esophageal cancer surgery, and provide basis for clinical diagnosis and treatment.Methods Patients were admitted to a hospital due to anastomotic fistula after esophageal cancer surgery between January 2012 and December 2015, microbial culture and antimicrobial susceptibility testing results of patients were retrospectively analyzed.Results 1 986 patients underwent radical resection of esophageal cancer within 4 years, 148 of whom developed anastomotic fistula, 104 (70.27％) were with positive microbial culture.A total of 197 pathogenic strains were isolated, 52(26.40％)and 145 (73.60％)strains were isolated from intrathoracic anastomotic fistula and cervical anastomotic fistula respectively;127 (64.47％)strains were gram-negative bacteria, the major were Pseudomonas aeruginosa, Klebsiella pneumoniae, and Acinetobacter baumannii, 62(31.47％) strains were gram-positive bacteria, the major were Staphylococcus aureus, Enterococcus spp., and Streptococcus viridans, 8 strains (4.06％) were fungi.49(47.12％) cases were with mixed pathogenic infection.The resistance rates of gram-negative bacteria to imipenem were 17.86％-47.62％, to polymyxin B was 0, resistance rates of Pseudomonas aeruginosa to other antimicrobial agents were all<50％, Klebsiella pneumoniae to piperacillin and aztreonam were both>70％, Acinetobacter baumannii to most antimicrobial agents were all>50.00％;resistance rates of gram-positive bacteria to clindamycin and tetracycline were both>50.00％, to linezolid, vancomycin, and teicoplanin were all 0, resistance rates of Staphylococcus aureus to penicillin, oxacillin, and ciprofloxacin were all>60％,resistance rate of Enterococcus spp.to quinupristin/dalfopristin was 100.00％.Conclusion Postoperative anastomotic fistula combined with infection can affect the prognosis of patients after esophageal cancer surgery, regular monitoring on distribution and drug resistance of pathogens can provide the basis for initial empirical treatment, and is conducive to the early treatment and rational use of antimicrobial agents.

Objective To investigate the risk factors of cervical esophagogastric anastomotic fistula after esophagectomy of esophageal cancer.Methods The retrospective case-control study was conducted.The clinicopathological data of 956 patients who underwent esophagectomy and cervical esophagogastrostomy from January 2012 to December 2016 in the First Affiliated Hospital of Zhengzhou University were collected.Patients underwent Sweet or Mckeown surgery.Observation indicators:(1) intra-and post-operative situations;(2) the risk factors analysis of cervical esophagogastric anastomotic fistula after esophagectomy;(3) follow-up situations.Follow-up using outpatient examination and telephone interview was performed to detect the esophagogastric anastomotic stenosis of patients up to February 2017.Measurement data with normal distribution were represented as the (x)±-s.Univariate analysis and comparison of count data were done using the chi-square test or Fisher exact probability method.Multivariate analysis was done using the Logistic regression model.Results (1) Intra-and post-operative situations:all the 956 patients underwent successful operations,including 107 with Sweet operation and 849 with Mckeown operation.Of 956 patients,336 received thoracotomy and 620 received thoracoscopic surgery.Tumors located in upper,middle and lower esophagus were respectively detected in 143,627 and 186 patients.Operation time,volume of intraoperative blood loss and number of lymph node dissected in 956 patients were (274 ± 67) minutes,(210 ± 167) mL and 18 ± 11,respectively.Of 956 patients,117 had cervical esophagogastric anastomotic fistula,with an incidence of anastomotic fistula of 12.24％ (117/956).Of 117 patients with cervical esophagogastric anastomotic fistula,2 had early stage fistula,110 had middle stage fistula and 5 had later stage fistula;12 were cured by two-tube method (stomach tube and nutrition tube),24 were cured by three-tube method (stomach tube,nutrition tube and chest tube or mediastinal tube),43 were cured by open neck incision dressing,15 were cured by fistula cavity drainage and 17 were cured by esophageal stent implantation.Sixteen patients died in hospital postoperatively,including 6 with cervical esophagogastric anastomotic fistula and 10 without cervical esophagogastric anastomotic fistula.Duration of hospital stay of 956 patients was (16± 11)days,and durations of hospital stay of patients with and without cervical esophagogastric anastomotic fistula were (39± 19) days and (13±6) days.Postoperative pathological examinations:873,9 and 74 patients were respectively diagnosed with squamous cell carcinoma,adenocarcinoma and other types of cancer.TNM staging:stage 0,Ⅰ,Ⅱ,Ⅲ,Ⅳ and unidentified stage were respectively detected in 135,110,325,376,1 and 10 patients.(2) The risk factors analysis of cervical esophagogastric anastomotic fistula after esophagectomy:univariate analysis showed that gender,age,history of diabetes,surgical method,tubular stomach production,operation time,postoperative pulmonary infection and postoperative aspirating sputum through fiberbronchoscope were risk factors affecting cervical esophagogastric anastomotic fistula after esophagectomy,with statistically significant differences (x2 =4.179,6.174,4.427,4.377,6.266,7.057,55.036,51.806,P＜ 0.05).Multivariate analysis showed that tubular stomach production,postoperative pulmonary infection and aspirating sputum through fiberbronchoscope were independent risk factors affecting cervical esophagogastric anastomotic fistula after esophagectomy,with statistically significant differences (OR =1.922,2.907,2.323,95％ confidence interval:l.203-3.070,1.682-5.023,1.235-4.370,P＜0.05).(3) Follow-up situations:908 of 956 patients were followed up for 2-62 months,with a median follow-up time of 28 months.During the follow up,21 of 111 patients with cervical esophagogastric anastomotic fistula were complicated with cervical esophagogastric anastomotic stenosis,59 of 797 patients without cervical esophagogastric anastomotic fistula were complicated with cervical esophagogastric anastomotic stenosis,showing a statistically significant difference in cervical esophagogastric anastomotic stenosis (x2-16.803,P＜0.05).Conclusion Tubular stomach production,postoperative pulmonary infection,postoperative aspirating sputum through fiberbronchoscope are independent risk factors affecting cervical esophagogastric anastomotic fistula after esophagectomy.

Objective To investigate the effect of liver kinase B1(LKB1) on the radiosensitivity of subcutaneous xenograft tumor of lung cancer H460 cells in nude mice.Methods Human lung cancer H460 cells were implanted into female nude mice (BALB/c-nu) to establish a subcutaneous xenograft tumor model of lung cancer.A total of 24 female nude mice in which the model was successfully established were equally and randomly divided into four groups:pEGFP-Ctrl plasmid (empty vector plasmid) group, irradiation (IR)+pEGFP-Ctrl plasmid group, pEGFP-LKB1 plasmid (overexpressing LKB1) group, and IR+pEGFP-LKB1 plasmid group.The growth of xenograft tumors was observed and the tumor inhibition rate and enhancement factor (EF) were calculated.The expression of LKB1 in each group was measured by immunohistochemistry and Western blot to analyze the relationship between LKB1 and radiosensitivity.Results Compared with the pEGFP-Ctrl plasmid group, the IR+pEGFP-Ctrl plasmid group, pEGFP-LKB1 plasmid group, and IR+pEGFP-LKB1 plasmid group showed varying degrees of inhibition of tumor growth, particularly in the IR+pEGFP-LKB1 plasmid group, and the tumor inhibition rates were 31.30%, 14.78%, and 43.48%, respectively.The EF of LKB1 in the IR+pEGFP-LKB1 plasmid group was 1.18.The immunohistochemistry and Western blot showed that LKB1 could be effectively expressed in the pEGFP-LKB1 plasmid group and IR+pEGFP-LKB1 plasmid group, but not in the other two groups.Conclusions The subcutaneous xenograft tumor model of human lung cancer H460 cells has been successfully established in nude mice.LKB1 has a radiosensitizing effect on the subcutaneous xenograft tumor of lung cancer H460 cells in nude mice.

Objective: To investigate clinicopathologic features and prognosis of adenoid cystic carcinoma (ACC) involving external auditory meatus. Methods: The clinical presentation and follow-up data of 63 patients with ACC of external auditory canal were collected from January 2006 to February 2017 at PLA General Hospital and Hainan Branch of PLA General Hospital. The clinicopathologic features and prognostic factors of external auditory canal ACC were analyzed. Results: (1) There were 28 males and 35 females and the average age of the first diagnosis was 48.9 years (22-81 years). The tumors showed cribriform pattern in 35 cases (15 cases of late stage), tubular pattern in 14 cases (8 cases of late stage), and solid pattern in 14 cases (9 cases of late stage). Cases with solid pattern was relatively more frequent than that of cribriform pattern and tubular pattern, but the difference was not statistically significant (P>0.05). (2) The average follow-up time was 62.4 months (2-228 months) in the 57 available cases. Among the 33 cases with recurrence, 18 cases had local recurrence and 15 cases had distant metastasis. The mean recurrence time was 40.6 months (2-204 months). Nine patients died of ACC: 2 cases in early stage (died at 48 and 102 months after the first treatment), 7 cases in late stage and 57 with (died at 9, 30, 32, 60, 72, 94 and 228 months). (3) Among the 37 patients with perineural invasion, there were 21 cases of cribriform pattern, 4 cases of tubular pattern and 12 cases of solid pattern; the number of cases in early stage and late stage were 15 and 22, respectively; and the differences were statistically significant (P<0.05). In addition, 31 cases had otalgia among the 37 patients with perineural invasion, where differences were not significant (P>0.05). (4) Thirty of 45 cases with tumor resection or partial resection of temporal bone had recurrence, whereas 3 of 12 cases of tumor combined with superficial lobectomy of parotid gland had recurrence. The difference was statistically significant (P<0.05). Postoperative adjuvant radiotherapy was given in 19 cases, including 7 cases of early stage (2 cases of recurrence), and 12 cases of late stage (8 cases of recurrence), among which there was no significant difference (P>0.05). Conclusions ACC occurring in external auditory canal frequently recurs. Superficial parotid lobectomy at the first operation is necessary to prevent tumor recurrence. Postoperative adjuvant radiotherapy has certain curative effect on patients with early stage tumor, but it does not affect the recurrence rate. Patients at late stage are more prone to perineural invasion than those in early stage. In addition, cribriform and solid patterns are more common that tubular pattern, and there is no significant correlation between perineural invasion and otalgia.

Objective To explore the application value of the fusiform tube stomach in the digestive tract reconstruction after thoracoscopic and laparoscopic radical resection of esophageal carcinoma.Methods The retrospective cohort study was conducted.The clinicopathological data of 96 patients with thoracic esophageal cancer who were admitted to the First Affiliated Hospital of Zhengzhou University between November 2016 and May 2017 were collected.All the patients underwent thoracoscopic and laparoscopic radical resection of esophageal carcinoma,45 using thin tubular stomach and 51 using fusiform tube stomach for digestive tract reconstruction were respectively allocated into the tubular stomach group and fusiform stomach group.Observation indicators:(1) intra-and post-operative situations;(2) postoperative complications;(3) detection of gastric hemodynamics;(4) follow-up situations.Follow-up using outpatient examination and telephone interview was performed to detect patients' survival up to November 2017.Measurement data with normal distribution were represented as-x±s and comparison between groups was analyzed using the t test.Measurement data with skewed distribution were described as M (range) and comparison between groups was analyzed using the rank sum test.Ordinal data and categorical variables were respectively done using the independent-sample Wilcoxon rank-sum test and chi-square test.Paired experimental data were analyzed by the Friedman test and Wilcoxon test.Results (1) Intra-and post-operative situations:96 patients underwent successful thoracoscopic and laparoscopic radical resection of esophageal carcinoma.The number of closers,time of postoperative mediastinal tube removal,thoracic stomach anteroposterior diameter by postoperative CT examination,number of slices at maximal width of thoracic stomach by postoperative CT examination and duration of hospital stay were respectively 4 (range,3-5),14 days (range,11-45 days),28.35 mm (range,9.96-75.70 mm),0.56 (range,0.33-2.13),16 days (range,12-62 days) in the tubular stomach group and 4 (range,2-4),12 days (range,10-16 days),45.80 mm (range,17.36-89.77 mm),1.10 (range,0.47-2.15),14 days (range,12-61 days) in the fusiform stomach group,with statistically significant differences between groups (Z=4.525,4.240,-3.796,-4.604,2.154,P＜0.05).(2) Postoperative complications:cases with postoperative cervical anastomotic fistula,thoracic gastric fistula,grading Ⅰ-Ⅱ and Ⅲ-Ⅳ of Clavien-Dindo classification were respectively 4,5,32,13 in the tubular stomach group (1 with a combination cervical anastomotic fistula and thoracic gastric fistula) and 0,0,47,4 in the fusiform stomach group,with statistically significant differences between groups (x2 =9.937,7.266,P＜0.05).Patients with complications were improved by symptomatic treatment.(3) Detection of gastric hemodynamics:hemodynamic values of gastric antrum,gastric body and gastric fundus that was detected by non-contact laser Doppler line imaging were respectively 314 PU (range,294-320 PU),252 PU (range,242-259 PU),206 PU (range,194-223 PU) in self-control status of 7 patients and 295 PU (range,277-314 PU),255 PU (range,244-267 PU),219 PU (range,199-233 PU) in tubular stomach model and 277 PU (range,263-300 PU),216 PU (range,201-235 PU),199 PU (range,176-207 PU) in fusiform stomach model,with statistically significant differences among groups (x2 =10.286,14.000,10.286,P＜0.05).There were statistically significant differences in the hemodynamic values of gastric antrum,gastric body and gastric fundus between self-control status and fusiform stomach model (Z=-2.028,-2.384,-2.197,P＜0.05),between self-control status and tubular stomach model (Z =-2.371,-2.371,-2.201,P＜0.05) and between fusiform stomach model and tubular stomach model (Z =-2.201,-2.366,-2.366,P＜0.05).(4) Follow-up situations:among 96 patients,90 were followed up for 6-12 months,with a median time of 8 months.During the follow-up,1 patient in the tubular stomach group died of tumor recurrrence,and no patient died in the fusiform stomach group,with no statistically significant difference between groups (x2 =1.264,P ＞ 0.05).Conclusion Compared with the thin tubular stomach,the fusiform tube stomach can reduce the incidences of postoperative fistula and pulmonary complications and shorten duration of hospital stay after the thoracoscopic and laparoscopic radical resection of esophageal carcinoma,and hemodynamics of the fusiform tube stomach is superior to that of thin tubular stomach.