Objective To investigate the effect of hydroxyapatite nanoparticles (nHAP) mediated human telomerase re-verse transcriptase (hTERT) RNA interference of A549 human lung cancer cells in vitro. Methods The nHAP were synthe-sized by the homogeneous precipitation method. The structure of the nanoparticles was observed under transmission electron mi-croscope. The nHAP were prepared using nltrasonication and Na_2CO_3 and modified with poly-L-lysine (PLL) at pH 7. 4. The transfection of pGenesil-hTERT into A549 was divided into four groups as follows: nHAP-PLL group mediated by hydroxyapatite nanoparticles modified with poly-L-lysine ( nHAP-PLL), liposome group mediated by Lipefectamine, nHAP group mediated by hydroxyapatite nanoparticles and control group. The growth ability of cells was assayed with methyl thiazolyl tetrazolium meth-od. The expression level of hTERT protein was examined by Western blotting. Flow cytometry was used to detect the apeptosis ratio of A549 cells line. Results Under transmission electron microscope, the synthesized product presented needle-like and well dispersed particles with evenly distributed sizes of (15-20) nm × (60-80) nm. The proliferation of A549 cells of nHAP-PLL group, liposome group and nHAP group were obviously inhibited as compared with the control group (P < 0.05 ).The inhibition rate of nHAP-PLL group was more than the other groups. There was a significant difference inhibition rate be-tween the nHAP-PLL group compared with the liposome group and nHAP group (P <0.05 ). The level of hTERT protein hada similar varietal tendency with the result of proliferation of each group. Flow cytometry showed the apoptasis ratio of nHAP-PLL group, liposome group, nHAP group and control group was (28.1±1.4)%, (19.2±1.3)%, (10.9±1.2)% and (0.3±0.2 ) %, respectively. There was a significant difference in apoptosis ratio between the nHAP-PLL group, liposome group and nHAP group compared with control group( P < 0.05 ). Conclusion A549 human lung cancer cells overexpreas hTERT and this may be a target for inhibiting proliferation of A549. Hydroxyapatite nanoparticles can induce apeptosis of ASA9 cells in vitro. Hydroxyapatite nanoparticles modified with poly-L-lysine can effectively combine and protect DNA and mediate gene transfection to A549, it can mediate human telomerase reverse transcriptase RNA interference of A549 cells and inhibit the pro-liferation of ,4549 in vitro.

Objective To compare survival according to the extent of lymph node dissection in patients with thoracic esophageal squamous cell carcinoma.To identify the subgroups of patients that could get survival benefit from three-field lymph node dissection.Methods Between January 1999 and December 2007,1551 patients with thoracic esophageal squamous cell carcinoma received esophagectomy plus three-field lymph node dissection (3 FL) (n =1131) or two-field lymph node dissection (2FL) (n =420).We retrospectively analyzed the clinical characteristics and patterns of lymphatic spread of thoracic esophageal squamous cell carcinoma.Survival rates between 3FL and 2FL were compared using the Kaplan-Meier method and Log-Rank test.Multivariate analysis were also performed to assess the element which affect the survival in 3FL and 2FL group by Cox regression.Results No significant differences in age,gender and depth of tumor invasion were found between 3 FL group and 2FL group.The 3FL group included more patients with upper thoracic esophageal tumors(17.6％ vs.9.8％) and patients with lymph node metastasis(LNM) (62.7％ vs.52.9％).Cox-proportional multivariate analysis showed that extent of lymph node dissection(3FL vs 2FL) was a significant prognostic factor in overall survival; 3 FL was beneficial for patients with upper thoracic esophageal tumors(P =0.002,5-year survival rate 53.2％ vs.34.1％).The 3FL group in patients with middle/lower thoracic esophageal tumors who had no LNMs(N0) had better 5-year survival than the 2FL group(5-year survival rate 77.5％ vs.70.7％),but no significant differences were found (P =0.235).or; Among patients with middle/lower thoracic esophageal tumors who had 1-6 LNMs (N1-N2),3 FL was beneficial for patients with mediastinum LNMs (P =0.006,5-year survival rate 41.1％ vs.32.8％) For patients with ≥7 LNMs(N3),cervical lymphadenectomy did not show additional survival benefits.Conclusion Our findings suggest that extent of lymph node dissection(3FL vs 2FL) is a significant prognostic factor for thoracic esophageal squamous cell carcinoma.3FL offers survival benefit over 2FL in patients with upper thoracic esophageal tumors or patients with middle/lower thoracic esophageal tumors who have 1-6 LNMs with mediastinum lymph node metastasis.

Objective To investigate the relationship of nodal skip metastasis(NSM) and clinicopathological factors of middle thoracic esophageal squamous cell carcinoma patients.Methods Between January 1999 and December 2007,695 patients with middle thoracic esophageal squamous cell carcinoma who had lymph node metastasis were reviewed.All patients received McKeown esophagectomy.We retrospectively analyzed the clinical characteristics and NSM status.Survival rates were compared using the Kaplan-Meier method and Log-Rank test.Multivariate analysis were also performed to assess the element which affect the survival by Cox regression.Results NSM were present in 226 (32.5％) patients.No significant differences in age,gender,tumor differentiation and extent of lymph node dissection depth of tumor invasion were found between skip metastasis group and continuous metastasis group.The NSM group included more patients with earlier T stage and N stage.Univariate analysis displayed that NSM was beneficial for patients with middle thoracic esophageal tumors (P ＜ 0.001).Cox-proportional multivariate analysis showed NSM was not a significant prognostic factor in overall survival.The overall survival did not differ according to NSM status in subgroups with different N stage.T1-2 patients,no significant difference of 5-year survival rate was found between skip metastasis group and continuous metastasis group(P =0.059).T3-4 patients,significant difference of 5-year survival rate was found between skip metastasis group and continuous metastasis group(P =0.001).NSM patients were then separated into 3 groups based on the extent of metastasis lymph nodes:both cervical and abdominal NSM (n =45,19.9％),cervical NSM (n =120,53.1％) and abdominal NSM (n =61,27.0％).The number of metastasis lymph nodes was significantly different among the three groups.No survival differences were observed among the three groups.Conclusion NSM is more frequently in the earlier stage compared to continuous metastasis.Three field lymphadenectomy can reduce the recurrence of T3-4 patients,and improve the survival rate of five years.The presence of NSM does not predict prognosis.

Objective To analyze the possible prognostic factors of thoracic-esophageal cancer patients with lymph node (LN) metastasis after esophagectomy plus three-field lymphadenectomy.Methods 590 esophageal cancer patients with LN metastasis after esophagectomy plus three-field lymphadenectomy were recruited from Jan.1993 to Mar.2007,and the prognostic factors and causes of postoperative failure were analyzed.Results Five-year survival in the whole sample was 29.6％.While the 5-year survival in different subgroups with 1-2,3-6 or ＞ 7 metastatic LNs were 41.2％,22.2％ and 7.0％ (x2 =62.158,P ＜0.0001),respectively.Univariate analysis showed that tumor site,disease length,T stage,number of metastatic LN and the seventh edition of AJCC staging system were prognostic factors.Multivariate analysis indicated that tumor site and number of metastatic LN were two independent prognostic factors.Conclusion Tumor site and number of metastatic LN were independent prognostic factors influencing the outcome of esophageal cancer.

Objective To compare the pulmonary complications between the double lumen tube and the single lumen tube,and to determine whether there are objective advantages of one over the other in one-lung ventilation during minimally invasive esophagectomy(MIE).Methods From January 2012 to November 2013,165 patients with esophageal cancer received combined thoracoscopic and laparoscopic esophagectomy with anastomosis in the neck.All clinical data were retrospectively reviewed.Results Between the single lumen tube and the double lumen tube:patients with pulmonary infection is 11 (16.42％),34(34.69％) (P =0.010).The average intubation time is(1.45 ±0.22)min,(6.53 ±0.59) min,P =0.000.The number of harvested lymph nodes of total is (42.76 ± 18.11) and (34.32 ± 15.80),P =0.002.The number of harvested lymph nodes of the cervix and the left laryngeal recurrent nerve chain was (3.19 ± 2.53) and (1.30 ± 2.14),P =0.000.Conclusion In the minimally invasive esophagectomy,single lumen tube is simpler and easier than the double lumen tube,and with the low incidence of postoperative pulmonary complications,at the same time there are more advantageous in the meditational lymph nodes cleaning.

Objective:To explore the role of NKG2D ligand MHC-I related molecule A (MICA) in chemotherapy combined with NK cell immunotherapy in patients with advanced esophageal cancer after surgery. Methods:A total of 90 patients with esophageal cancer from Fujian Provincial Tumor Hospital were divided into three groups after surgery:40 patients of chemotherapy alone, 25 patients of chemotherapy combined with NK cell therapy with negative expression of MICA (MICA-group), and 25 patients of chemotherapy combined with NK cells therapy with positive expression of MICA (MICA+group). The efficacy was then compared. Results:Compared with the chemotherapy alone and MICA-groups, the positive rates of CD3+, CD4+T cells, NK cells, and the CD4+/CD8+ratio in peripheral blood from MICA+group were higher than those before treatment (64.2%± 6.4%vs. 51.3%± 5.6%, 39.8%± 8.2%vs. 29.5%± 3.2%, 25.3%± 2.1%vs. 16.4%±4.3%, 1.4%± 0.5%vs. 1.1%± 0.7%;P<0.05). Meanwhile, the levels of T-reg cells were lower than those before treatment (6.3%± 4.5%vs. 17.3%± 2.4%, P<0.05). No significant difference was observed between the disease control rate and response rate. Chemotherapy-induced neutropenia and peripheral neurotoxicity symptoms were significantly improved, and time to progression (TTP) and overall survival (OS) were significantly prolonged (P<0.05). No statistically significant difference was observed between the chemotherapy alone group and MICA-group (P>0.05). Conclusion:Treatment with chemotherapy and autologous NK cells on patients with advanced esophageal carcinoma and MICA positive expression can be safely transfused with only minor side effects and can effectively improve a patient's immune system, quality of life, and survival.

Objective To analyze the characteristic of serum proteins in non-small cell lung cancer (NSCLC) patients, establish serum markers pattern for the diagnosis of NSCLC. Methods Surface enhanced laser desorption ionization time of flight mass spectormetry ( SELDI-TOF-MS) technology was used to analyze serum samples. Bio-marker Pattern Software (BPS) was used to detect the protein peaks. Results Sixteen significantly different pro-tein peaks were found in serum samples in NSCLC patients and healthy controls. Eight up-regulated protein peaks and eight down-regulated protein peaks ( P < 0. 001 ) were identified in serum samples of NSCLC patients. Three up-regulated protein peaks(P <0. 05) were identified in serum samples of patients of NSCLC with smoking history. Two up-regulated protein peaks(P <0. 01) were identified in serum samples of patients of squamous carcinoma comparing with adenocarcinoma. No significantly different protein peak was found in serum samples of NSCLC patients at different clinical stages . Conclusion SELDI - TOF - MS technology can identify different protein peaks and so function as a diagnostic tool with high sensitivity and specificity.

Objective To explore the relevance of expression of MHC class Ⅰ-related molecules A (MICA) molecule and NK cells immunotherapy in esophageal cancer patients after operation.To analyze the significance of MICA expression in NK cell immunotherapy.Methods 100 patients of esophageal cancer were divided into 3 group,surgical alone group,MICA negative with NK therapy group (MICA-group) and MICA positive with NK therapy group (MICA+ group).The immunity indicators and tumor markers including the levels of CD3+,CD4+ T cells,ratio of CD4+/CD8+, NK cells,Treg cells,the levels of Th1/Th2/Th17 cytokine,the antibody IgA,IgM,IgG and the tumor markers of CEA,SCC,CA199,CYFRA21-1 were detected before treatment and after treatment 60 days.Results The positives rates of CD3+,CD4+ T cells,NK cells and the ratio of CD4+/CD8+ in peripheral blood from MICA+ patients group were higher than those of before treatment [(68.3±7.6) ％ vs (56.2±4.1) ％,(39.8±8.2) ％ vs (30.8±4.7) ％,(22.2±4.7) ％ vs (18.7±5.5) ％,(1.49±0.30) vs (1.15±0.61),P ＜ 0.05],meanwhile the levels of Treg cells was lower than those of before treatment [(8.1± 4.0) ％ vs (13.4±4.5) ％,P ＜ 0.05].There was no statistical significant difference of positive rate of CD8+ T cells [(26.9±6.2) ％ vs (27.8±7.1) ％,P ＞ 0.05].The levels of Th1 cytokin (IL-2,IFN-γand TNF-α) increased and Th2 cytokin (IL-4,IL-6 and IL-10) decreased after treatment.The level of Th17 cytokine was not different significantly (P ＞ 0.05).The content of IgA,IgM,IgG in MICA+ group were effectively improved after treatment.The tumor markers CEA,SCC,CA199,CYFRA21-1 had no statistically change before and after treatment.Conclusion The results indicate that NK cells immunotherapy can enhance the cellular immunity and humoral immunity of MICA positive esophageal cancer patients after operation.

Objective:To explore the association of microsatellite polymorphism of MICA gene with susceptibility to esophageal cancer.Methods: PCR-STR microsatellite genotyped technique was used to detect the polymorphism of MICA in Exon 5 in 103 cases of esophageal cancer and 84 cases of normal controls.Constructed of eukaryotic expression vector in esophageal carcinoma with high frequency of occurrence of the MICA allele.NK cells killing effect to 293T cells after alleles MICA transfected were assayed by LDH and the effect on target was 20∶1.ELISA was used to test supernatants sMICA of 293T cell after transfected.Results: Identified five allelic genes in MICA Exon 5 with esophageal cancer.Each allele and its frequency respectively were:MICA-A4(9.71%),MICA-A5(22.3%),MICA-A5.1(40.8%),MICA-A6(15.5%),MICA-A9(11.7%).MICA-A5.1 showed significant difference comparison with the control group.After 293T cell line was transfected MICA allele,MICA-A5.1 group was less sensitive to NK cytotoxicity compared to other groups[(30.4±6.3)%,P<0.05].The secretion of soluble MICA increased(135.7±6.2)pg/ml.Conclusion: Esophageal cancer was relevent with the MICA-A5.1 polymorphism of MICA Exon 5 alleles.Its risk is higher than other alleles.

Objective To analyze the characteristic of serum proteins in non-small cell lung cancer (NSCLC) patients,establish serum markers pattern for the diagnosis of NSCLC. Methods Surface enhanced laser desorption ionization time of flight mass spectormetry(SELDI-TOF-MS) technology was used to analyze serum samples. Biomarker Pattern Software (BPS) was used to detect the protein peaks. Results Sixteen significantly different protein peaks were found in serum samples in NSCLC patients and healthy controls. Eight up-regulated protein peaks and eight down-regulated protein peaks (P