Objective To investigate whether modification with IL-21 and CCL19 enhances killing and tumor-infiltrating efficiency of NKP30 CAR-T cells in lung cancer.Methods The modified IL-21-CCL19 NKP30 CAR-T cells expressing IL-21 and CCL19 fusion gene was constructed based on NKP30 CAR-T cells and stimulated with CD3CD28 antibodies and IL-2.The immunophenotype and migration of the cells in the presence of IL-21 were investigated using flow cytometry and migration experiments.Lactate dehydrogenase(LDH)release and sphere formation assays were used to assess the killing and infiltration capabilities of CAR-T cells,and the secretion levels of IFN-γ,IL-21 and CCL19 were determined with enzyme-linked immunospot assay(ELISPOT)and ELISA.A zebrafish model bearing HCG-27 cell xenograft was established by microinjection of the tumor cells into the yolk sac followed 24 h later by injection of the immune cells at the same site,and the fluorescence signals were captured using a fluorescent microscopy.Results The NKP30 ligand B7H6,which was almost undetectable in normal tissues and blood cells,was highly expressed(over 90%)in lung cancer cells.Compared with NKP30 CAR-T cells and conventional T cells,IL-21-CCL19 NKP30 CAR-T cells exhibited stronger proliferative and migration capabilities with the formation of central memory T cells.The reduced expressions of CTLA4 and PD1 in the constructed cells resulted in enhanced killing efficiency against lung cancer cells accompanied by significantly increased production of IFN-γ,IL-21 and CCL19.In the zebrafish models,CAR-T cells exhibited stronger cytotoxicity and proliferative abilities than typical T cells,but these differences were not statistically significant between the two CAR-T cells.Conclusion Modification of NKP30 CAR-T cells with IL-21 and CCL19 facilitates their access into solid tumors for more effective tumor cell killing while producing a large number of memory T cells.

Objective To investigate whether modification with IL-21 and CCL19 enhances killing and tumor-infiltrating efficiency of NKP30 CAR-T cells in lung cancer.Methods The modified IL-21-CCL19 NKP30 CAR-T cells expressing IL-21 and CCL19 fusion gene was constructed based on NKP30 CAR-T cells and stimulated with CD3CD28 antibodies and IL-2.The immunophenotype and migration of the cells in the presence of IL-21 were investigated using flow cytometry and migration experiments.Lactate dehydrogenase(LDH)release and sphere formation assays were used to assess the killing and infiltration capabilities of CAR-T cells,and the secretion levels of IFN-γ,IL-21 and CCL19 were determined with enzyme-linked immunospot assay(ELISPOT)and ELISA.A zebrafish model bearing HCG-27 cell xenograft was established by microinjection of the tumor cells into the yolk sac followed 24 h later by injection of the immune cells at the same site,and the fluorescence signals were captured using a fluorescent microscopy.Results The NKP30 ligand B7H6,which was almost undetectable in normal tissues and blood cells,was highly expressed(over 90%)in lung cancer cells.Compared with NKP30 CAR-T cells and conventional T cells,IL-21-CCL19 NKP30 CAR-T cells exhibited stronger proliferative and migration capabilities with the formation of central memory T cells.The reduced expressions of CTLA4 and PD1 in the constructed cells resulted in enhanced killing efficiency against lung cancer cells accompanied by significantly increased production of IFN-γ,IL-21 and CCL19.In the zebrafish models,CAR-T cells exhibited stronger cytotoxicity and proliferative abilities than typical T cells,but these differences were not statistically significant between the two CAR-T cells.Conclusion Modification of NKP30 CAR-T cells with IL-21 and CCL19 facilitates their access into solid tumors for more effective tumor cell killing while producing a large number of memory T cells.

Resection is one of the most important treatments for esophageal squamous cell carcinoma, and routine postoperative follow-up is an effective method for early detection and treatment of recurrent metastases, which can improve patients' quality of life and prognosis. This consensus aims to provide a reference for colleagues responsible for postoperative follow-up of esophageal squamous cell carcinoma patients in China, and further improve the standardization of the diagnosis and treatment of esophageal squamous cell carcinoma.

Objective:To investigate the epidemiological characteristics and current status of surgical management for esophageal cancer in China.Methods:A national database was setup through a network platform. The clinical data of esophageal cancer treated by surgery was collected from 70 major hospitals in China between January 2009 and December 2014.Results:Complete data of 8 181 cases of esophageal cancer patients who underwent surgery were recorded in the database and recruited in the analysis. Among them, 6 052 cases were male and 2 129 were female, the average age was 60.5 years.The epidemiological investigation results showed that 148 cases (1.8%) had history of psychological trauma, 7 527 cases (92.0%) were lower social economic status, 5 072 cases (62.0%) were short of fresh vegetables and fruits, 6 544 cases (80.0%) ate rough food frequently, 3 722 cases (45.5%) drank untreated water directly from lake or river or shallow well, 3 436 cases (42.0%) had a unhealthy eating habit, including habits of eating food fast (507 cases, 6.2%), eating hot food or drinking hot tea/soup (998 cases, 12.2%), eating fried food (1 939 cases, 23.7%), 4 410 cases (53.9%) had the habits of smoking cigarettes and 2 822 cases (34.5%) drank white wine frequently.The pathological results showed that 7 813 cases (95.5%) were squamous cell carcinoma, 267 cases were adenocarcinoma (3.3%), 25 cases were adenosquamous cell carcinoma (0.3%) and 50 cases were small cell carcinoma (0.6%). A total of 1 800 cases (22.0%) received preoperative neoadjuvant therapy due to locally advanced disease or difficulty of resection. The esophagectomies were performed through left thoracotomy approach in 5 870 cases (71.8%), through right chest approach in 2 215 cases (27.1%), and the remain 96 cases (1.2%) received surgery though other approaches.A total of 8 001 cases (97.8%) underwent radical resection, the other 180 cases (2.2%) received palliative resection. The 30-day postoperative mortality rate was 0.5%, the overall ≥ grade Ⅱ postoperative complication rate was 11.6% (951 cases). The 1-yr, 3-yr, and 5-yr overall actual survival rates were 82.6%, 61.6%, and 52.9%, respectively.Conclusions:The data analysis of the national database for esophageal cancer shows that bad eating habits or eating rough food without enough nutrients, lower social and economic status, drinking white wine and smoking cigarettes frequently may be correlated with tumorigenesis of esophageal cancer. However, strong evidences produced by prospective observation studies are needed. Overall, the long-term survival of esophageal cancer patients has been improved gradually due to the application of advanced surgical techniques and reasonable multimodality treatment.

Objective:To investigate the epidemiological characteristics and current status of surgical management for esophageal cancer in China.Methods:A national database was setup through a network platform. The clinical data of esophageal cancer treated by surgery was collected from 70 major hospitals in China between January 2009 and December 2014.Results:Complete data of 8 181 cases of esophageal cancer patients who underwent surgery were recorded in the database and recruited in the analysis. Among them, 6 052 cases were male and 2 129 were female, the average age was 60.5 years.The epidemiological investigation results showed that 148 cases (1.8%) had history of psychological trauma, 7 527 cases (92.0%) were lower social economic status, 5 072 cases (62.0%) were short of fresh vegetables and fruits, 6 544 cases (80.0%) ate rough food frequently, 3 722 cases (45.5%) drank untreated water directly from lake or river or shallow well, 3 436 cases (42.0%) had a unhealthy eating habit, including habits of eating food fast (507 cases, 6.2%), eating hot food or drinking hot tea/soup (998 cases, 12.2%), eating fried food (1 939 cases, 23.7%), 4 410 cases (53.9%) had the habits of smoking cigarettes and 2 822 cases (34.5%) drank white wine frequently.The pathological results showed that 7 813 cases (95.5%) were squamous cell carcinoma, 267 cases were adenocarcinoma (3.3%), 25 cases were adenosquamous cell carcinoma (0.3%) and 50 cases were small cell carcinoma (0.6%). A total of 1 800 cases (22.0%) received preoperative neoadjuvant therapy due to locally advanced disease or difficulty of resection. The esophagectomies were performed through left thoracotomy approach in 5 870 cases (71.8%), through right chest approach in 2 215 cases (27.1%), and the remain 96 cases (1.2%) received surgery though other approaches.A total of 8 001 cases (97.8%) underwent radical resection, the other 180 cases (2.2%) received palliative resection. The 30-day postoperative mortality rate was 0.5%, the overall ≥ grade Ⅱ postoperative complication rate was 11.6% (951 cases). The 1-yr, 3-yr, and 5-yr overall actual survival rates were 82.6%, 61.6%, and 52.9%, respectively.Conclusions:The data analysis of the national database for esophageal cancer shows that bad eating habits or eating rough food without enough nutrients, lower social and economic status, drinking white wine and smoking cigarettes frequently may be correlated with tumorigenesis of esophageal cancer. However, strong evidences produced by prospective observation studies are needed. Overall, the long-term survival of esophageal cancer patients has been improved gradually due to the application of advanced surgical techniques and reasonable multimodality treatment.

Objective To investigate the pattern of lymph node metastasis and the long-term survival in patients with superficial thoracic esophageal squamous cell carcinoma(ESCC) with T1 status.Methods 176 patients with pathologically confirmed superficial ESCC with T1 status who underwent Mckeown esophagectomy between January 1999 and January 2010 were retrospectively enrolled.Tumor invasion is classified according to the Japanese Association of esophageal cancer classification standard.Epithelial layer and mucosa(m) divided into epithelial layer(M1),lamina propria mucosa(M2),muscularis mucosa(M3),submucosa(M3).Submucosa(Tsm) were average divided into submucous upper 1/3 (SM1),middle 1/3 (SM2) and lower 1/3 (SM3).Node metastatic pattern of different tumor invasion and the prognostic indication is investigated.Results The rate of node metastasis in patients with SM invasion is significantly higher than those with m invasion (28.2％ vs.4.4％,P =0.001);the 5-year survival rate of patients with SM invasion is significantly higher than those with m invasion (91.4％ vs.75.8％,P =0.048).The rate of node metastasis in patients with M1,M2,and M3 invasion is 0,0,and 6.2％,respectively;the rate of node metastasis in patients with SM1,SM2,and SM3 invasion is 20.0％,29.4％,and 33.3％,respectively,with the 5-year survival rate of 86.5％,77.4％,and 67.3％,respectively.The highest rate of lymph node metastasis in the neck region,mainly in the right cervical paraesophageal lymph nodes(101R).The rate of cervical lymph node metastasis in patients with SM1,SM2,and SM3 invasion is 15.0％,20.6％ and 24.6％,respectively.The rate of 101R metastasis in patients with SM1,SM2,and SM3 invasion is 10％,8.8％ and 24.6％,respectively.Univariate analysis showed that there is no significant difference between lymph node metastasis rate and patient age,gender,tumor differentiation and tumor location,and vascular invasion and tumor invasion depth (ml-3/SM1-3) difference is statistically significant.Nodal status is proved as the independent prognostic factor(HR =2.127,95％ CI =1.219-3.713).The 5-year survival of patients with and without node metastasis is 64.7％ and 83.5％ (P =0.005).Conclusion The rate of node metastasis of SM invasion cohort is significantly higher than M invasion cohort.There was no significant difference in lymph node metastasis rate between SM1,SM2 and SM3.Lymph node metastasis(N staging) is an independent prognostic factor for stage T1 thoracic ESCC.Compared with the thoracic and abdominal lymph nodes,the cervical lymph node metastasis rate is higher,and concentrated in 101R.

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.

Differences in operative procedure and knowledge of esophageal cancer exist among surgeons from different countries and regions. There is controversy in the surgical treatment of esophageal cancer, especially in the extent of lymphadenectomy. Until now, results of the three-field lymphadenectomy and two-field lymphadenectomy are mostly reported by retrospective studies from Japan and China. Three-field lymphadenectomy has been initiated in Fujian Provincial Cancer Hospital since 1990s. After evaluating our database, we found that three-field was superior to two-field lymphadenectomy in terms of long-term survival for patients with upper thoracic esophageal cancer, whereas for those with middle or lower thoracic esophageal cancer, the survival benefit of three-field lymphadenectomy was reduced. Therefore, we propose to perform three-field lymphadenectomy for upper thoracic esophageal cancer. In middle or lower thoracic esophageal cancer, we suggest to perform modified two-field lymphadenectomy in most cases, and three-field lymphadenectomy in selective cases. Video-assisted two-field lymphadenectomy is feasible. Based on the national condition of China, we advise to perform thoracic duct removal only in patients with posterior mediastinal or peri-ductus node metastasis to achieve curative effect.
Carcinoma, Squamous Cell ; China ; Esophageal Neoplasms ; classification ; mortality ; surgery ; Esophagectomy ; methods ; Humans ; Lymph Node Excision ; methods ; Lymphatic Metastasis ; Mediastinum ; Retrospective Studies ; Survival Rate ; Thoracic Duct ; surgery ; Thoracic Neoplasms ; classification ; surgery ; Thoracic Surgery, Video-Assisted

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 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.