Retrospect of 2019: focus on the research progress of gastric neoplasm / 中华胃肠外科杂志

Gastric carcinoma, gastrointestinal stromal tumor and gastric neuroendocrine tumor are the most common gastric neoplasms. A series of researches in 2019 showed that the safety and efficacy of laparoscopic gastrectomy in the treatment of both early and advanced gastric cancer patients are similar to open surgeries, providing a high-level evidence-based medical basis for the promotion of laparoscopic surgery in the treatment for gastric cancer. In multidisciplinary treatment and perioperative chemoradiotherapy, major research results have also been published, and clinical researches in China are gradually gaining international recognition and attention. Although the application of targeted therapy and immunotherapy has made progress, the first-line therapy after gastric cancer surgery has not been established. In the field of gastrointestinal stromal tumors, laparoscopic surgery has gradually been recognized, and surgical treatment of patients with advanced drug resistance still has its value. In terms of gastric neuroendocrine tumors, the latest researches showed that surgical methods should be selected according to tumor characteristics, and gastric adenocarcinoma with neuroendocrine components may have a worse prognosis.

The shift of the treatment strategy of gastrointestinal stromal tumor from the surgical perspective / 中华外科杂志

As tumors originated from mesenchymal tissue, gastrointestinal stromal tumors (GIST) has its own typical history. For the idea of treatment for GIST at different historical periods, the role and value of surgery for the treatment of GIST keep changing. Laparoscopy and endoscopy will have the role they deserved. With the understanding of pathogenesis of GIST, targeted chemotherapy will be more and more accurate and individualized. How to improve the overall therapeutic effect of GIST, especially for the patients with the high risk and drug-resistance, is the dilemma and challenges for the surgeons.

Elabela promotes differentiation of wharton’s jelly-derived mesenchymal stem cells into cardiomyocyte-like cells / 中国组织工程研究

BACKGROUND: Elabela is a new type of endogenous receptor of APJ discovered in recent years. It is widely distributed in the adult cardiovascular system and has a certain influence on cardiovascular diseases. However, the effect of Elabela on the differentiation of stem cells into cardiomyocytes and the expression of APJ in cardiomyocyte differentiation has not been studied yet. OBJECTIVE: To investigate the effect of Elabela on the differentiation of Wharton’s jelly-derived mesenchymal stem cells into cardiomyocytes. METHODS: The frozen mesenchymal stem cells were resuscitated. 5-Azacytidine was used to induce Wharton’s jelly-derived mesenchymal stem cells to differentiate into cardiomyocytes when the cell confluence reached 80%-90%. After 24 hours, the medium was replaced by low-glucose medium containing Elabela and 10% fetal bovine serum in the experimental group, and by low-glucose medium containing 10% fetal bovine serum in the control group. At 7, 14, 21, and 28 days after induction, cell morphology was observed. The total RNA and total protein of each group were collected. The myocardial specific markers Nkx2.5, cTnT and Connexin 43 mRNA and protein expression levels were detected by real-time fluorescent quantitative PCR and western blot assay. The expression of APJ in the induced cardiomyocytes was detected by real-time fluorescent quantitative PCR and flow cytometry. RESULTS AND CONCLUSION: (1) The expression levels of myocardial specific markers Nkx2.5, cTnT and Connexin 43 mRNA and protein were higher in the experimental group than in the control group in all stages of differentiation, and the expression of APJ was also higher in the experimental group than in the control group. (2) In summary, Elabela plays a certain promoting role in the differentiation of Wharton’s jelly-derived mesenchymal stem cells into oriented cardiomyocytes. Elabela, as another agonist of APJ, can promote the expression of APJ during the induced cell differentiation.

Evidence-based medicine should be emphasized to improve the quality of gastrointestinal surgery / 中华消化外科杂志

With the further application of evidencebased medicine and the change of disease spectrum,gastrointestinal surgery is facing the novel challenge and opportunity.Systemic quality control of surgery will have positive effects on the subject development.Absence of high level of evidence,the popularization of mini-invasive surgery should be carried out cautiously.Standardization of definition of complications and unified management will be helpful to the clinical researches,prevention and treatment of complications.Conception and implementation of multidisciplinary team and enhanced recovery after surgery will provide every patient individualized and standard treatment.

Future trends and prospect of surgery of gastrointestinal cancer in China / 中华外科杂志

With the development of medical concepts and technology, the surgical treatment of gastrointestinal cancer is gradually moving towards a new era. Chinese gastrointestinal surgeons are building up our own high-level evidence in evidence-based medicine, with tremendous clinical trials carrying out in gastrointestinal cancers. Besides, standardized procedure of diagnosis and treatment should be promoted. More personalized schemes are needed. The model of multidisciplinary team can be more widely and deeply applied. And the concept of enhanced recovery after surgery should be more operable. To keep pace with the times, the gastrointestinal surgeons have to seek for innovative technology and new ideas.

Clinical research status of laparoscopic total gastrectomy in China / 中华胃肠外科杂志

Many retrospective and cohort studies have shown that laparoscopic total gastrectomy(LTG) has advantages of less trauma, quicker recovery and better incision than open total gastrectomy, and is not inferior to laparotomy in safety and the short- and long-term outcomes, so it has been widely applied. However there is still a lack of high level evidence-based basis. At present, several prospective multicenter clinical studies are being carried out in China, Japan and South Korea which have high incidence of gastric cancer to demonstrate the safety and feasibility of LTG and evaluate its long-term outcomes as well. A multicenter randomized controlled clinical trial comparing the safety of laparoscopic and open radical gastrectomy(CLASS-02 Research) conducted by Professor Sun Yihong, Zhongshan Hospital of Fudan University, is under way. The subjects of this study were the patients with clinical stage I(, the lesion locating in the gastric body, gastric fundus, or the esophagogastric junction (not involving the cardia dentate line), who were expected to be treated with total gastrectomy. The primary outcome measures were early operative morbidity and mortality and the secondary outcome measures were early postoperative recovery course and postoperative hospital stay. It is believed that results of the study will provide a high level evidence-based basis for the safety of LTG. The ongoing JCOG-1401 study in Japan aims to assess the feasibility and safety of LTG and LAPG for the treatment of early gastric cancer. LTG (Roux-en-Y reconstruction) or LAPG (double-tract or jejunal interposition reconstruction) will be performed in patients with clinical stage I( gastric cancer in this study. The primary endpoint of the study is the incidence of anastomotic leakage, and the secondary endpoints are proportion of patients converted to open surgery, the time to the first flatus, analgesic requirement, postoperative body temperature, the overall survival and relapse-free survival. The follow-up time is at least 5 years, and the results will also effectively evaluate the long-term efficacy of LTG and LAPG in the treatment for early gastric cancer. No.10 lymph nodes dissection of D2 total gastrectomy is a major difficulty in LTG. Japanese JCOG-0110 study has showed that for proximal gastric cancer patients without involvement of the greater curvature of the stomach, routine splenectomy for No.10 lymph nodes dissection is not recommended. In this regard, Professor Huang Changming, Union Hospital of Fujian Medical University, led the ongoing CLASS-04 study to evaluate the safety, feasibility and clinical efficacy of laparoscopic No.10 lymph node dissection with spleen preservation for patients with locally advanced gastric cancer locating in the upper part of the stomach. The ongoing CLASS-02 and JCOG-1401 studies will answer whether LTG is suitable for the treatment of early gastric cancer. We believe that in the absence of high level evidence-based basis, LTG should be applied cautiously to advanced gastric cancer, and LTG for early gastric cancer patients should also be operated by experienced surgeons.

Trend of development and prospect of gastrointestinal surgery in era of minimally invasive / 中华外科杂志

The development and popularization of minimally invasive treatment is gradually evolving in the course of human fighting against disease. Minimally invasive surgery has been widely accepted and carried out because of the advantages of smaller trauma, less pain, little impact on the body and physiological metabolism of the body. The evidence-based medicine makes it important to conduct clinical trials to confirm the technical and oncological safety. With the development of minimally invasive technologies, including three-dimensional laparoscopic, natural orifice transluminal endoscopic surgery, robotic surgery system, minimally invasive surgery are providing more opportunities and challenges for gastrointestinal diseases. The standard conduction of minimally invasive surgery in gastroenterology disease makes it important to improve surgical procedure safety and treatment effects.

Multidisciplinary team improves the comprehensive quality of colorectal surgeons / 中华胃肠外科杂志

Colorectal cancer is a systemic disease that requires multidisciplinary treatment. The comprehensive quality of colorectal surgeon directly impacts on the efficacy of diagnosis and treatment of colorectal cancer. Multidisciplinary teams help surgeons enhance their ability of evidence-based medicine, improve the quality of main specialty, expand the knowledge of other specialty, enhance the doctor-patient communication, and increase the research level. Thus, multidisciplinary teams can improve the comprehensive quality of colorectal surgeons.

Strategies for prevention and treatment of postoperative complications of gastric cancer / 中华胃肠外科杂志

Postoperative complications after gastric cancer surgery has their own specificity and complexity, and the strategies for prevention and treatment should be of equal emphasis on both theory and technology. Based on the knowledge and familiarity with different postoperative complications, to efficiently prevent them, it is not only necessary to strengthen the training of acknowledged operative strategy, smooth and precise surgical techniques, but also to address the importance of overall preoperative assessment for patients, to treat the basic diseases, and to improve and correct their general conditions. Combining with the concept and basic protocol of enhanced recovery after surgery (ERAS), it is preferred to work out an individualized perioperative preventing strategy for patients who have high risk factors of specific postoperative complications. After the operation, to guarantee intensive and individual managements for patients, to catch early abnormal signs, then to make early and precise diagnosis, and to do timely response and accurate treatments, including timely and proper re-operations, can improve the efficacy of complications and promote the recovery of patients as soon as possible.

Efficacy analysis of targeted therapy combined with surgery in the treatment of recurrent and metastatic gastrointestinal stromal tumor / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To investigate the efficacy of targeted therapy combined with surgery in the treatment of recurrent and metastatic gastrointestinal stromal tumor(GIST).</p><p><b>METHODS</b>Clinicopathological and followed-up data of 318 patients with recurrent and metastatic GIST admitted in Zhongshan Hospital between January 2000 and December 2015 were analyzed retrospectively. According to different treatment methods, the patients were divided into four groups: surgery group (operation alone, 44 cases), target therapy group (imatinib alone, 108 cases), target therapy combined with surgery group (imatinib plus operation, 139 cases), other therapy group (chemotherapy, Chinese medicine and others, 27 cases). The progression-free survival (PFS) and overall survival (OS) of four groups were compared.</p><p><b>RESULTS</b>The baseline informations, such as age, gender, primary site, et al, were not significantly different (all P>0.05), but the recurrent and metastatic site was significantly different among 4 groups (P=0.000). The medial PFS of surgery group, target therapy group, target therapy combined with surgery was 16(95%CI: 4.9 to 27.0) months, 44 (95%CI: 30.9 to 57.1) months, 35 (95%CI: 26.5 to 43.5) months, respectively, and the latter 2 groups had significantly longer PFS than surgery group(P=0.000), while no significant difference was found between target therapy group and target combined with surgery group (P=0.251). The median OS of surgery group, target therapy group, target therapy combined with surgery, and other therapy group was 24 (95%CI: 9.0 to 39.0) months, 69(95%CI: 40.8 to 97.2) months, 92(95%CI: 78.0 to 106.0) months, 12(95%CI: 9.5 to 14.5) months. Target therapy group and target therapy combined with surgery group had significantly longer OS than surgery and other therapy groups (P=0.000), while the target therapy combined with surgery group had significantly longer OS than target therapy group(P=0.028).</p><p><b>CONCLUSION</b>Target therapy combined with surgery can prolong the survival of recurrent and metastatic GIST patients.</p>

Clinicopathological analysis of 80 patients with duodenum gastrointestinal stromal tumors / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To explore the clinicopathological characteristics, efficacy, and prognostic factors for patients with duodenum gastrointestinal stromal tumor(GIST).</p><p><b>METHODS</b>Clinicopathological and follow-up data of 80 patients with duodenum GIST in the Zhongshan Hospital from January 2000 to December 2013 were analyzed retrospectively.</p><p><b>RESULTS</b>There were 38 male and 42 female patients with a median age of 54 years. The major symptoms were upper alimentary tract hemorrhage and abdominal pain. Thirty-nine patients received local tumor excision, 18 patients underwent segmental duodenectomy, 23 patients were subjected to pancreaticoduodenectomy, all these operations were R0 resection. Thirty patients received imatinib treatment after operation, and 11 among them had metastasis relapse. Recurrence-free survival rates of 1-, 3-, and 5-years were 96.2%, 90.6%and 78.6% retrospectively. Overall survival rates of 1-, 3-, and 5-years were 100%, 98.3% and 96.1%. Multivariate Cox analysis showed tumor size >5 cm, mitotic count >5 mitosis/50 HPF and intermediate/high NIH risk classification were associated with an increased risk of recurrence. The significant difference was not detected between the limited resection group and pancreaticoduodenectomy group in OS and RFS.</p><p><b>CONCLUSIONS</b>Surgery is still the main treatment for duodenum GIST. The surgical program is mainly determined by the location and size of tumor. Imatinib therapy should be used if necessary.</p>

Standardized training of gastrointestinal surgeons should be emphasized / 中华胃肠外科杂志

The standardized training of residents and specialists has just been initiated, and the training for gastrointestinal surgeons also should be standardized. From my personal view, the following aspects should be addressed for standardization including basic theory, medical record, teaching ward-round, the ability of clinical skills and clinical research. After the establishment of systematic standard training and assessment, the sustained development of gastrointestinal surgery may be expected.

Clinicopathological features of small gastrointestinal stromal tumors / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To investigate the clinicopathological features of small gastrointestinal stromal tumors(GISTs) and to evaluate the efficacy of endoscopic therapy for GISTs.</p><p><b>METHODS</b>Clinicopathological and follow-up data of 418 patients with GISTs undergoing endoscopic therapy in the Zhongshan Hospital between January 2009 and July 2014 were analyzed retrospectively. All the cases were evaluated by the NIH risk classification and AIFP classification, and were grouped according to the tumor size and location. Nuclear atypia and mitotic count were used to evaluate the biological behavior of small GIST. Efficacy of endoscopic therapy was analyzed with follow-up data.</p><p><b>RESULTS</b>Out of 418 patients, GISTs located in the esophagus was 14(3.3%), in the stomach 389(93.1%), in the duodenum 5(1.2%), in the rectum 10(2.4%). A total of 412(98.6%) patients were mainly spindle cells, and mitosis was not found in 320(76.5%) patients. In 389 small stomach GIST, 245(58.6%) were in fundic region. Cases were divided into four groups according to the size and the result revealed the bigger the size, the more the mitotic count. Nuclear atypia in the 1.5-1.9 cm group was significantly higher compared to other groups. Cases were divided into four groups according to the location and the result revealed the mitotic count was not associated to the location. While the nuclear atypia of stomach GIST was significantly higher than that of esophageal GIST and the nuclear atypia of rectum GIST was significantly higher than that of other positions. The median follow-up was 32(4-69) months. One case(gastric fundus GIST, >1.5 cm) presented local recurrence 23 months after operation and underwent endoscopic resection again. No recurrence or metastasis was found in other patients.</p><p><b>CONCLUSIONS</b>Endoscopic resection technique is effective for small GISTs patients. The small GISTs with 0.4 cm diameter or less are often benign and should be followed up for long time. The small GISTs with 0.5 cm diameter or more possess the risk of malignancy, then surgical resection should be performed. Rectum small GISTs (except for 0.4 cm diameter or less) have worse biological behavior and should be removed.</p>

Is there a principle for digestive reconstruction after laparoscopic gastrectomy? / 中华胃肠外科杂志

The laparoscopic gastrectomy has been popularized world-wide, and more and more clinical trials have been designed and conducted. Nowadays, lymphadenectomy is not a technical problem for Chinese surgeons, and the type of digestive reconstruction is closely related with patients' quality of life. Besides the basic principle and indication, which both laparoscopic and open gastrectomy must follow, there are some technical characteristics of laparoscopic gastrectomy. So the maneuver and methods of reconstruction for laparoscopic gastrectomy is different from open surgery. Surgeons must carefully evaluate the clinical evidences and patient's conditions, individualize the optimal type of reconstruction, and conduct clinical trials.

Comparative study of laparoscopic and open surgery for gastric gastrointestinal stromal tumors / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To investigate the feasibility and short-term efficacy of laparoscopic resection of primary localized gastric gastrointestinal stromal tumors (GIST) by comparing with open surgery.</p><p><b>METHODS</b>Clinicopathological data of 167 gastric GIST patients undergoing operation in Zhongshan Hospital from June 2008 to December 2013 were retrospectively analyzed, among whom 55 received laparoscopic surgery and 112 underwent open surgery for primary local gastric GIST. Efficacy of different size and different location of GIST was compared between laparoscopic and open groups.</p><p><b>RESULTS</b>There was no conversion to open surgery in laparoscopy group. Compared with open surgery, laparoscopic resection for gastric GIST smaller than 5 cm or located at anterior wall, greater curvature, lesser curvature, was associated with similar operation time(P>0.05), but less blood loss, shorter post-hospital stay or flatus time(all P<0.05). The operative outcomes were similar between laparoscopic and open resection for gastric GIST bigger than or equal to 5 cm or located at posterior wall(all P>0.05), except the longer operation time in laparoscopy group(P<0.05). The incidence of postoperative complication did not differ between two groups. Laparoscopic group had 2 patients with gastroparesis and open group had 2 gastroparesis, 2 pulmonary infection, and 1 poor wound healing(all P>0.05), which all recovered after conservative treatment. During 7 to 84 months(median 35) of follow-up, no recurrence or hepatic metastasis was found in laparoscopy group, and 3 hepatic metastases in open group. There was no significant difference of recurrence-free survival between two groups(P>0.05).</p><p><b>CONCLUSION</b>Laparoscopic resection for gastric GIST is safe and effective in selected patients, especially for those with tumors smaller than 5 cm, or located at anterior wall, greater curvature, lesser curvature, whose short-term outcomes are better than open resection.</p>

Clinical control study of laparoscopic versus open surgery for rectal cancer / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To evaluate the safety and short-term outcomes of laparoscopic-assisted surgery for rectal cancer by comparing the efficacy of laparoscopy and open surgery.</p><p><b>METHODS</b>Clinical data of patients with rectal cancer treated by laparoscopy or open surgery in Zhongshan Hospital from April 2011 to June 2012 were analyzed retrospectively, and the clinical outcomes between the two groups were compared.</p><p><b>RESULTS</b>Ninety-six rectal cancer patients undergoing laparoscopic surgery(LS) were enrolled. A total of 216 rectal cancer patients underwent open surgery(OS). There was no operative death in both groups. In LS and OS group, the overall completion rates of TME were 86.4%(83/96) vs. 89.3%(193/216)(P>0.05) respectively, and the overall anal reservation rates were 78.1%(75/96) vs. 75.0%(162/216)(P>0.05) respectively. The mean distance to proximal resection margin and distal resection margin respectively were (10.3±4.1) cm vs.(10.0±4.3) cm(P>0.05) and (3.4±0.9) cm vs. (3.6±1.4) cm(P>0.05) respectively. The mean number of harvested lymph nodes respectively were (12.8±5.2) vs.(13.7±6.4)(P>0.05). Compared to OS, LS presented less blood loss [(98.0±28.7) ml vs. (175.0±41.0) ml, P<0.05], shorter postoperative hospital stay [(9.4±4.9) d vs.(11.6±6.2) d, P<0.05], quicker postoperative recovery of bowel function[(2.7±0.9) d vs. (3.4±0.9) d, P<0.05], shorter postoperative time to intake semi-solid[(3.7±1.2) d vs. (4.4±1.5) d, P<0.05], less postoperative complications(15.6% vs. 25.9%, P<0.05), but longer operative time[(155.7±48.4) min vs. (120.0±26.7) min, P<0.05]. Postoperative follow-up was 6 to 24 months, and the local recurrence of LS and OS was 2.1% and 2.3%(P>0.05).</p><p><b>CONCLUSION</b>Laparoscopic surgery can obtain the same radical efficacy for rectal cancer as compared to open surgery.</p>

Impact of the number of lymph nodes dissected on the prognosis of G3 advanced gastric cancer / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To explore the risk factors associated with the prognosis in poorly differentiated(G3) advanced gastric cancer (AGC) and the effect of number of harvested lymph nodes on the prognosis.</p><p><b>METHODS</b>Clinical data of 484 patients with G3 advanced gastric cancer undergoing gastrectomy combined with lymphadenectomy in Zhongshan Hospital from December 2002 to October 2007 were retrospectively analyzed. The χ(2) test or Fisher's exact probability method was used to test measurement data. Survival was calculated using Kaplan-Meier method. Clinicopathological factors such as age, invasion depth, number of harvested lymph nodes, lymph node metastasis, ratio of metastatic lymph node, lymphatic vessels involvement were analyzed using the Cox regression model.</p><p><b>RESULTS</b>Number of lymph node dissected, lymph node metastasis, ratio of metastatic lymph node and lymphatic vessels involvement were significantly affected by groups of 15 lymph node dissected in G3 AGC patients (P<0.05). Invasion depth, number of lymph node dissected, lymph node metastasis, ratio of metastatic lymph node and lymphatic vessels involvement were significantly independent predictors of survival in G3 AGC patients (all P<0.05). Patients with at least 20 lymph nodes had a better survival rate than those with less than 20(P<0.01).</p><p><b>CONCLUSIONS</b>Invasion depth, number of harvested lymph nodes, lymph node metastasis, ratio of metastatic lymph node and lymphatic vessels involvement are important factors influencing the prognosis of G3 AGC. At least 20 lymph nodes should be recommended to ensure the quality of lymphadenectomy for gastric cancer.</p>

Macrophage-derived soluble factors induce the epithelial-mesenchymal transition of HepG2 cells / 中华肝胆外科杂志

Objective This study investigates the epithelial-mesenchymal transition effects exerted on human liver cancer cells HepG2 by THP-1 derived macrophages.The roles of tumor-associated macrophages (TAMs) on liver cancer progression and its mechanisms were explored.Methods HepG2 cells were cultured with THP-1 derived macrophages to mimic the microenvironment of liver cancer.After the culture treatment,morphological changes of the liver cancer cells were observed.Decreased E-cadherin expression is a hallmark of epithelial-mesenchymal transition (EMT),and the Ecadherin protein variations in the HepG2 cells were detected by immunofluorescence (IF) and Westernblot.FlowCytomix was carried out to screen the cytokines in the supernanants of THP-1 cells and THP-1 derived macrophages.Results After culture with macrophages,HepG2 cells revealed a morphological change.These cells lacked epithelial morphology and became a spindle-like mesenchymal cell phenotype.Additionally,the E-cadherin protein expression was reduced dramatically as measured by IF and Western-blot.IL-8 and IL-1β expression in the supernatants were increased 40 and 20 times,respectively,after THP-1 cells were activated to macrophages (P＜0.01).TNF-α expression was increased 8 times (P =0.056).Conclusion THP-1 derived macrophages could induce EMT effects on HepG2 cells,which may relate to the increased secretion of IL-1β,IL-8 and TNF-α.

The immunogenicity of the new MUC1 DNA vaccine for pancreatic cancer / 中华肝胆外科杂志

Objective The development of cancer vaccines deserves experimentation,specifically the immunogenicity of the new MUC1 DNA vaccine for pancreatic cancer.Methods Three strategies were combined to optimize the new MUC1 DNA vaccine.The female C57BL/6 mice were immunized,through tibial muscle injection,with 100 μg of plasmid DNA of the recombinant plasmids (pIRES2-EGFP-3VNTR group,pIRES2-EGFP-3VNTR-C1-144 group,pIRES2-EGFP 3VNTR-mIL-18 group,pIRES2-EGFP-3VNTR-C1-144-mIL-18 group,n=5) for a total volume of 100 μl.Mice inoculated with the empty vector pIRES2-EGFP (EV group,n=5) and normal saline (NS group,n=5) were used as vector and blank controls,respectively.All the mice were immunized again every two weeks.Two weeks after the third immunization,all the mice were euthanized and spleen cells were separated for CTL cytotoxic assay.Results The specific cytolysis percentages of the four groups (pIRES2-EG-FP-3VNTR,pIRES2-EGFP-3 VNTR-C1-144,pIRES2-EGFP-3VNTR-mIL-18,pIRES2-EGFP 3VNTR-C1-144-mIL-18) expressing VNTR were higher than the EV and NS group with the effector/target cells ratio (E/T) from 80:1 to 20:1 (P＜0.05).Therefore,it showed a difference among the four groups.After the primary immunization,the OD450 of the serum antibody level specific to MUC1 began to increase in the four groups which contained the gene of VNTR3 (P＜0.05).This suggests that the recombinant plasmids could induce a specific antibody response to MUC1,and showed no remarkable difference among the four groups.IFN-γ serum cytokine among the four groups were higher than that of the EV and NS groups (P＜0.05).There was a significant difference of OD450 between the groups containing mIL-18 pIRES2-EGFP-3VNTR-mIL-18,pIRES2-EGFP-3VNTR-C1-144-mIL-18) and those not (pIRES2-EGFP-3VNTR,pIRES2-EGFP-3VNTR-C1-144,)(P＜0.05).Conclusions In conclusion,all of the four recombinant plasmids could induce MUC1 specific CTL and antibodies responses,and C1-144 and IL-18 could enhance the immunogenicity of plasmids.