Effects of oral immunonutritional supplement on 3-year disease-free survival in gastric cancer patients with pathological stage III after total gastrectomy (CRUCIAL): study protocol of a multicentre, randomised clinical trial.

INTRODUCTION: The nutritional status of patients with gastric cancer (GC) after total gastrectomy continues to deteriorate and lasts a long time after discharge, which is an independent risk factor for mortality. Recent guidelines have recommended appropriate nutritional support after discharge for cancer surgery patients with malnutrition or nutritional risk. The evidence on the efficacy of oral immunonutritional supplement (INS) and its effect on long-term disease-free survival (DFS) in patients with GC is limited. This study was designed to test the hypothesis that oral INS compared to diet alone may improve 3-year DFS of GC patients with pathological stage III after total gastrectomy (Nutrition Risk Screening 2002 score ≥3 at discharge). METHODS AND ANALYSIS: This is a pragmatic, open-label, multicentre, randomised controlled study. 696 eligible GC patients with pathological stage III after total gastrectomy will be randomised in a 1:1 ratio to oral INS group or normal diet group for 6 months. The primary endpoint is 3-year DFS after discharge. The following secondary endpoints will be evaluated: 3-year overall survival; unplanned readmission rate at 3 and 6 months after discharge; quality of life, body mass index and haematological index at 3, 6 and 12 months after discharge; incidence of sarcopenia at 6 and 12 months after discharge; and the tolerance to chemotherapy. The adverse events of oral INS will also be evaluated during the intervention. ETHICS AND DISSEMINATION: This study was approved by the ethics committee of Jinling Hospital, Nanjing University (number 2021NZKY-069-01). The present study may validate the effectiveness of oral immunonutritional therapy in improving 3-year DFS for GC patients with pathological stage III after total gastrectomy for the first time. The results of this trial will be disseminated in peer-reviewed journals and at scientific conferences. TRIAL REGISTRATION NUMBER: NCT05253716.

Comprehensive Multiomics Analysis Identified IQGAP3 as a Potential Prognostic Marker in Pan-Cancer.

Background: IQGAP3 has important function in cancer progression and has become a potential therapeutic target as a transmembrane protein. But its role in tumor immunity and pan-cancer was not systematically investigated. This study evaluated the potential role of IQGAP3 and clinical significance in pan-cancer through combined multiomics analysis. Methods: From Genotype Tissue Expression (GTEx) and The Cancer Genome Atlas (TCGA) databases, transcriptomic datasets were first obtained, and from Gene Expression Omnibus (GEO), expression profiling microarray data were acquired and integrated to systematically assess the expression differences and prognostic relevance of IQGAP3 in pancreatic cancer. Immunohistochemical data were obtained from Human Protein Atlas (HPA) to assess IQGAP3 protein expression differences, and exome data from TCGA were used to analyze IQGAP3 expression in relation to tumor mutational burden (TMB), microsatellite instability (MSI), and mutation. Additionally, we also analyzed the relationship between IQGAP3 expression and immune checkpoints, mismatch repair (MMR), and IQGAP3 relationship with methylation and copy number variation based on expression profiles. Results: Microsatellite instability (MSI), immune checkpoints, mismatch repair (MMR), and tumor mutational burden (TMB) all closely interacted with IQGAP3 mRNA. In addition, detailed relationships between the immune microenvironment and IQGAP3 mRNA as well as immune cell CD4+ Th2 and myeloid-derived suppressor cells (MDSCs) were determined. Mechanistically, IQGAP3 was involved in cytoskeleton formation, T cell receptor signaling pathways, DNA damage, cell cycle, P53 pathway, Fc gamma R-mediated phagocytosis, and apoptosis. Conclusion: IQGAP3 could serve as an effective prognostic biomarker for pan-cancer immune-related therapy.

Silencing of UBE2D1 inhibited cell migration in gastric cancer, decreasing ubiquitination of SMAD4.

BACKGROUND: Gastric cancer (GC) is the second leading cause of cancer-related deaths. Because it is hard to diagnose at early stage, the overall 5 years survival rate is lower than 25%. High migration is the main hallmark of malignant cells at advanced stage of GC. Thus, it is urgent to find biomarkers for early diagnosis and more effective therapy of GC. METHODS: In this study, lentivirus-mediated silencing and overexpression lentiviruses targeting the ubiquitin-conjugating enzyme E2 D1 (UBE2D1), transwell, wound healing, and pulmonary metastasis mouse model were applied to analyze the function of UBE2D1 in vitro and in vivo. Real-time PCR and immunohistochemistry were used to elucidate the level of UBE2D1 in GC samples. RESULTS: Silencing of UBE2D1 inhibited cell migration and the levels of epithelial-mesenchymal transition makers (MMP2 and MMP9) in AGS and MKN45 cells. Silencing of UBE2D1 inhibited cell metastasis in mouse model. On the contrary, UBE2D1 overexpression increased cell migration and the levels of MMP2 and MMP9 in MGC-803 cells. Further, silencing of UBE2D1 decreased the ubiquitination level of mothers against decapentaplegic homolog 4 (SMAD4), and the increase of cell migration induced by UBE2D1 overexpression could be reversed by SMAD4. CONCLUSION: Silencing of UBE2D1 inhibited cell migration through transforming growth factor ß (TGF-ß)/SMAD4 signaling pathway in GC.

Prognostic value of lymphovascular invasion in stage II colorectal cancer patients with an inadequate examination of lymph nodes.

BACKGROUND: Lymphovascular invasion (LVI) is defined as the presence of cancer cells in lymphatics or blood vessels. This study aimed to evaluate the prognostic value of LVI in stage II colorectal cancer (CRC) patients with inadequate examination of lymph nodes (ELNs) and further combined LVI with the TNM staging system to determine the predictive efficacy for CRC prognosis. Adjuvant chemotherapy (ACT) was then evaluated for stage II CRC patients with LVI positivity (LVI+). METHODS: In order to avoid the effects of different ACT regimens, among 409 stage II patients, we chose 121 patients who received FOLFOX regimen and the 144 patients who did not receive ACT as the object of study. LVI was examined by hematoxylin-eosin (HE) staining. Kaplan-Meier analysis followed by a log-rank test was used to analyze survival rates. Univariate and multivariate analyses were performed using a Cox proportional hazards model. Harrell's concordance index (C-index) was used to evaluate the accuracy of different systems in predicting prognosis. RESULTS: The LVI+ status was significantly associated with pT stage, degree of differentiation, tumor stage, serum CEA and CA19-9 levels, perineural invasion (PNI), tumor budding (TB), and KRAS status. The 5-year overall survival (OS) rate of stage II patients with < 12 ELNs and LVI+ was less than stage IIIA. Multivariate analyses showed that LVI, pT-stage, serum CEA and CA19-9 levels, PNI, TB, and KRAS status were significant prognostic factors for stage II patients with < 12 ELNs. The 8th TNM staging system combined with LVI showed a higher C-index than the 8th TNM staging system alone (C-index, 0.895 vs. 0.833). Among patients with LVI+, the ACT group had a significantly higher 5-year OS and 5-year disease-free survival (DFS) than the surgery alone (SA) group (5-year OS, 66.7% vs. 40.9%, P = 0.004; 5-year DFS, 64.1% vs. 36.3%, P = 0.002). CONCLUSIONS: LVI is an independent prognostic risk factor for stage II CRC patients. Combining LVI with the 8th TNM staging system improved the predictive accuracy for CRC prognosis. ACT in stage II CRC patients with LVI+ is beneficial for survival.

Spontaneous hemopneumothorax after laparoscopy: a case report and literature review.

BACKGROUND: Spontaneous hemopneumothorax (SHP) is defined as the accumulation of >400 mL of blood in the pleural cavity in association with spontaneous pneumothorax. This rare clinical disorder may be life-threatening. CASE PRESENTATION: A 71-year-old woman presented with a 1-month history of recurrent bloody stool, and electronic colonoscopy suggested a rectal mass. Laparoscopic radical resection of rectal cancer was performed. Two days later, she developed chest tightness, shortness of breath, and slight pain in the left chest. Emergency chest radiography revealed mild left pneumothorax and pleural effusion. SHP was suspected and a thoracic drain was inserted. However, the patient developed hemorrhagic shock 3 hours after drainage. She underwent emergency video-assisted thoracic surgery (VATS), which revealed left lung tip rupture with bleeding and adhesive band fracture at the top of the left thoracic cavity. The ruptured lung tissue was removed and electrocoagulation at the adhesion band was performed for hemostasis. The patient was discharged on postoperative day 11. At the time of this writing, she had developed no SHP recurrence or any other complications. CONCLUSIONS: This case shows that conservative treatment may have serious consequences in patients with SHP. Thus, chest X-ray examination and VATS should be performed in patients with SHP.

Programmed death-ligand 1 and survival in colorectal cancers: A meta-analysis.

BACKGROUND: Programmed death-ligand 1 (PD-L1) is a programmed death 1 (PD-1) ligand that plays a pivotal role in the inhibition of the T-cell-mediated immune response. The expression of PD-L1 is associated with the prognosis and clinical outcomes of multiple tumors. However, the prognostic value of PD-L1 overexpression in colorectal cancer is still controversial. In this study, we sought to clarify this by presenting a meta-analysis of relevant studies. METHODS: Databases including PubMed, Web of Science, and EMBASE were systematically searched for studies concerning the expression of PD-L1 and survival in colorectal cancer. The reported hazard ratios (HR) with 95% confidence intervals (CI) of overall survival, disease-free survival, and recurrence-free survival in the included studies were analyzed by fixed effects/random effects models. RESULTS: Fifteen studies involving 3078 patients with colorectal cancer were included in our meta-analysis. Overexpression of PD-L1 was found to be associated with poor overall survival (HR 1.83; 95% CI 1.21, 2.79; P = 0.005) and poor recurrence-free survival (HR 2.78; 95% CI 1.43, 5.42; P = 0.003). However, no correlation was found between PD-L1 overexpression and poor disease-free survival (HR 1.23; 95% CI 0.83, 1.82; P = 0.305). Overexpression of PD-L1 indicating poor survival held true across different geographical areas, sample sizes, analysis types, sources of HRs, and cell types. CONCLUSION: Overexpression of PD-L1 is associated with worse prognosis in patients with colorectal cancer and can guide physicians in the application of PD-1/PD-L1 immune checkpoint-targeted therapy.

High expression of IQGAP3 indicates poor prognosis in colorectal cancer patients.

BACKGROUND: The oncogene IQ motif-containing GTPase activating protein 3 (IQGAP3) is ubiquitously overexpressed in several human cancers. This study was designed to explore the expression and role of IQGAP3 in colorectal cancer. METHODS: We first assessed the IQGAP3 expression level in colorectal cancer. The correlation of IQGAP3 expression with the clinicopathological characteristics and prognosis was then assessed. At last, we investigated the function of IQGAP3 in colorectal cancer by knocking down its expression in colorectal cancer cell lines. RESULTS: Consistent with the conclusions drawn from The Cancer Genome Atlas database, IQGAP3 was upregulated in colorectal cancer at the tissue level and cellular level. Based on immunohistochemistry results of the tissue microarrays, we demonstrated that higher expression of IQGAP3 was associated with higher tumor node metastasis stage (P = 0.005), higher incidence of lymph node metastasis (P = 0.004), and shorter overall survival (P = 0.022). Knockdown of IQGAP3 in colorectal cancer cell lines remarkably decreased their proliferation and migration abilities. CONCLUSION: Our data provide evidence that IQGAP3 significantly promote malignant progression of colorectal cancer and could serve as a potential therapeutic target.

Lymphoepithelioma-like gastric carcinoma treated with partial gastrectomy: Two case reports.

Lymphoepithelioma-like gastric carcinoma (LELGC) is a rare type of gastric cancer characterized by intense lymphocytic infiltration of the stroma. LELGC is associated with Epstein-Barr virus infection and has a favorable prognosis compared with other types of gastric carcinoma. The clinical symptoms of LELGC are usually similar to those of conventional gastric carcinoma. The diagnosis of LELGC is established based on pathological, histological and immunohistochemical findings. The present report describes the cases of two patients with LELGC who underwent esophagogastroduodenoscopy and computed tomography scans prior to surgery. The two patients accepted surgical treatment, and postoperative pathological and immunohistochemical analyses confirmed LELGC. Neither patient experienced local recurrence or distant metastasis during the postoperative period.

Clinical value of detecting IQGAP3, B7-H4 and cyclooxygenase-2 in the diagnosis and prognostic evaluation of colorectal cancer.

BACKGROUND: The IQ-motif-containing GTPase-activating protein (IQGAP) family comprises three members, IQGAP1, IQGAP2 and IQGAP3. IQGAP3 is the latest addition to the family. This study mainly investigated the novel marker IQGAP3 at serum and tumor tissue levels compared with the markers B7-H4 and cyclooxygenase-2 (COX-2) in patients with colorectal cancer (CRC) and in healthy individuals, aiming to evaluate the diagnostic and prognostic value of IQGAP3 for CRC. MATERIALS AND METHODS: Serum samples were collected prior to any therapy in 118 CRC patients and as part of a routine examination in 85 healthy individuals. Serum IQGAP3, B7-H4 and COX-2 levels were measured using commercially available ELISA kits. Immunohistochemistry was performed to detect the IQGAP3, B7-H4 and COX-2 in tumor tissues and normal para-carcinoma tissues. The receiver operating characteristics (ROC) curve and the area under the curve (AUC) were used to evaluate and compare the diagnostic value of different serum tumor markers. Univariate and multivariate analyses were performed to identify the prognostic risk factors for CRC. RESULTS: IQGAP3, B7-H4 and COX-2 showed low or high expression in tumor tissues while no expression in normal para-carcinoma tissues. Serum levels of IQGAP3 in CRC group were significantly higher than those in healthy control group (P < 0.001). The IQGAP3 AUC was 0.799, while the B7-H4 AUC was 0.795 and the COX-2 AUC was 0.796. IQGAP3 seemed to be superior to B7-H4 and COX-2 in detecting CRC, with the highest sensitivity among the three markers. Multivariate analysis showed that T stage, N stage, differentiation degree, TNM stage and both serum and tissue IQGAP3, B7-H4 and COX-2 levels were significant prognostic factors for CRC. CONCLUSIONS: IQGAP3 has a better diagnostic efficacy than B7-H4 and COX-2 in detecting CRC and it has value in predicting the prognosis of patients with CRC.

Comparison of the 8th union for international cancer control lymph node staging system for gastric cancer with two other lymph node staging systems.

The log odds of positive lymph nodes (LODDS) and the metastatic lymph node ratio (MLR) staging systems have previously been demonstrated to exhibit advantages compared with the tumor-node-metastasis (TNM) staging system in predicting the prognosis of gastric cancer. The current study compared the prognostic significance of the newest Union for International Cancer Control Node classification with the LODDS and MLR staging systems. From September 2010 to December 2012, all medical records for patients with gastric cancer at the Third Affiliated Hospital of Soochow University were retrospectively analyzed and the clinicopathologic characteristics were reviewed. Cut-off points were selected to divide the patients with gastric cancer into different groups. Univariate and multivariate analyses were performed to identify the prognostic risk factors for gastric cancer. The Harrell's concordance index (C-index) was adopted to compare the prognostic value of the three staging systems. A total of 877 patients with gastric cancer who met the inclusion criteria were analyzed in the current study. The patients were classified according to the three MLR subgroups as follows: MLR0 (MLR=0), MLR1 (00.5). Based on multivariate analysis, LODDS, MLR and pathological node (pN) stage could significantly predict survival rates of patients with gastric cancer. According to the C-index, the LODDS staging system more accurately predicted the 5-year overall survival for patients with gastric cancer compared with the other two staging systems. In summary, the current study has identified that LODDS may be superior to the MLR and pN staging systems in predicting the prognosis of patients with gastric cancer. However MLR may exhibit advantages compared with LODDS for patients who have undergone adequate lymphadenectomies.

Which Combination Treatment Is Better for Spinal Metastasis: Percutaneous Vertebroplasty With Radiofrequency Ablation, 125I Seed, Zoledronic Acid, or Radiotherapy?

BACKGROUND: Percutaneous vertebroplasty (PVP) can not only alleviate pain but also restore mechanical stability with injection of bone cement, whereas it exhibits a poor effect on antitumor activity. But through combinations with other therapies, it may be possible to achieve the maximum effect in clinical treatment. Thus, this study is designed to assess the clinical efficacy of PVP separately combined with 4 ways for spinal metastasis (SM) treatment. STUDY QUESTION: Which combination treatment is better for spinal metastasis, percutaneous vertebroplasty with radiofrequency ablation, I seed, zoledronic acid or radiotherapy? STUDY DESIGN: A total of 169 patients with SM were retrospectively recruited and randomly assigned to 4 groups to receive 4 different ways separately: 49 patients (group A) received PVP plus I seed, 51 (group B) received PVP plus radiofrequency ablation (RFA), 38 (group C) underwent PVP plus zoledronic acid (ZA), and 31 (group D) underwent PVP plus radiotherapy (RT). MEASURES AND OUTCOMES: All of them underwent routine examinations before operation. Visual analog scale (VAS), World Health Organization (WHO) Pain Relief, and ODI were applied to evaluate pain relief and motor function. RESULTS: PVP plus RT achieved the best efficacy in relieving pains, with the highest WHO Pain Relief (P < 0.05). The PVP plus RFA exhibited lowest ODI, suggesting the best outcome after treatment (P < 0.05). The PVP plus I showed the lowest VAS score, but it was the worst to improve the routine exercise ability and relieve pains from patients. The PVP plus ZA presented higher VAS and ODI (P < 0.05). CONCLUSIONS: PVP combined with I seed exhibited the best clinical efficacy in terms of VAS, PVP combined with RT was the best choice in terms of WHO Pain Relief, and PVP combined with RFA showed the best effect in terms of ODI for the treatment of SM.

Should Pyloric Lymph Nodes Be Dissected for Siewert Type II and III Adenocarcinoma of the Esophagogastric Junctions: Experience from a High-Volume Center in China.

BACKGROUND: The optimal extent of lymph node (LN) dissection remains controversial in adenocarcinoma of the esophagogastric junction (AEG), especially in Siewert types II and III. The aim of this study was to analyze clinicopathological characteristics of patients with Siewert type II and III AEGs to clarify whether pyloric (no. 5 and no.6) lymphadenectomy is essential in these patients. METHODS: A retrospective analysis was performed in the Third Affiliated Hospital of Soochow University from September 2008 to December 2012, and clinicopathological characteristics on all patients with Siewert type II and III AEGs, who underwent curative total gastrectomy with lymphadenectomy were collected. The index of estimated benefit from lymph node dissection (IEBLD) was used to evaluate the efficacy of lymph node dissection of no. 5 and no. 6. Both clinicopathological characteristics and IEBLDs were set as the standards in the assessment of the value of pyloric lymph nodes dissection. RESULTS: A total of 216 patients with AEG (Siewert type II: 141, Siewert type III: 75) were included into the study. Type III AEG had a larger tumor size and relatively advanced T stage compared to Type II AEG. The 5-year overall survival (OS) rates in type II and type III AEGs were almost similar (type II 50.4% vs. type III 46.7%, p = 0.782). There was a very low incidence of pyloric lymph nodes metastases in type II AEG (no. 5 is 1.4% and no. 6 is 0.7%). Hence, the IEBLDs of no. 5 and no. 6 lymph node were negligible regardless of the T stage and tumor differentiation. In type III AEG, metastasis rates of no. 5 and no. 6 lymph node were 9.3 and 5.3%, respectively. The IEBLDs of no. 5 and no. 6 lymph node were 2.7 and 1.3, respectively. CONCLUSIONS: Based on the IEBLDs of pyloric lymph nodes, dissection of no. 5 and no. 6 lymph nodes were worthwhile for Siewert type III AEG but not essential for Siewert type II AEG.

Liver receptor homologue 1, a novel prognostic marker in colon cancer patients.

Liver receptor homologue 1 (LRH-1) is an orphan nuclear receptor that is highly expressed in a variety of cancer tissues, promotes tumor cell proliferation and metastasis, and is involved in the tumor cell cycle and apoptosis. The aim of the present study was to assess the association between the expression of LRH-1 and the prognosis of patients with colon cancer. Immunohistochemistry was used to detect the expression of LRH1 in 128 cases of colon cancer and adjacent tissues. The 5-year survival rate was obtained from telephone follow-up data, outpatient review and through access to medical records. Positive expression of LRH-1 was found in 108/128 colon cancer samples, compared with 17/128 normal tissues. Statistical analysis showed that positive LRH-1 expression was significantly associated with clinical pathological stage, depth of invasion and lymph node metastasis. The overall survival (OS) rate of patients with positive LRH-1 expression was significantly lower than that of patients with low expression. Multivariate analysis showed that LRH-1 expression could be used as an independent predictor of OS. In conclusion, the present findings suggest that LRH-1 may serve an important role in the development and progression of colon cancer, with potential value as a prognostic molecular marker that could be used to assist in the diagnosis and evaluation of colon cancer. LRH-1 may become a target for novel therapies for patients with colon cancer.

Distinctive profiles of tumor-infiltrating immune cells and association with intensity of infiltration in colorectal cancer.

Tumor-infiltrating immune cells are heterogeneous and consist of characteristic compartments, including T helper (Th)1 and regulatory T (Treg) cells that exhibit distinctive biological functions. The present study investigated the profile of infiltrating immune cells from surgically removed tumor tissues from patients with colorectal cancer. The characteristic transcription factors of Th1 and Th2 cells, Treg cells, Th17 cells and T follicular helper (Tfh) cells were analyzed. The results demonstrated that a marked increased number of Treg cells presented in tumor infiltrates when compared with non-tumor adjacent tissues. An increased number of Th1 and Tfh cells existed in tumor infiltrates compared with non-tumorous adjacent tissues, while the infiltration of Th17 and Th2 cells was similar between tumor and non-tumor adjacent tissues. Furthermore, there were an increased number of Treg cells in tumors with low infiltration compared with those with high infiltration. The expression of CXC motif chemokine (CXC) receptor 3, CXC ligand (CXCL)L9 and CXCL10 was significantly increased on infiltrating T cells in tumors with high infiltration as compared with those with low infiltration. Macrophages exhibited a dominant M2 phenotype in tumor infiltrates of colorectal cancer, whereas a balanced M1 and M2 phenotype presented in macrophages from the peripheral blood. In vitro stimulation of macrophages isolated from tumor tissue of colorectal cancer with granulocyte macrophage colony-stimulating factor and lipopolysaccharide did not drive to an inflammatory phenotype. The results provide insights into the pattern of immune cell infiltration in Chinese patients with colorectal cancer. It may be beneficial that patients with colorectal cancer are screened for the defined profile along with the expression of CXCL9 and CXCL10 in order to achieve better efficacy in clinical applications of immune-based therapy, including anti-programmed cell death protein 1 therapy.

Prognostic value of pretreatment serum carbohydrate antigen 19-9 level in patients with colorectal cancer: A meta-analysis.

BACKGROUND: Carbohydrate antigen 19-9 (CA 19-9) is one of the most frequently used tumor markers for gastrointestinal cancer, particularly for diagnostic purposes. However, its value in predicting prognosis remains controversial. In this study, we sought to clarify this by conducting a meta-analysis of relevant studies. METHODS: We systematically searched several databases, including PubMed, EMBASE and Web of Science for articles pertaining to the relationship between pretreatment serum CA 19-9 levels and prognosis in patients with colorectal cancer (CRC). The reported hazard ratios (HR) of overall survival (OS), disease-free survival (DFS), pooled progression-free survival (PFS) and recurrence-free survival (RFS) in the analyzed studies were compared by fixed effects/random effects models. RESULTS: Seventeen studies involving 6434 patients with CRC were included in our meta-analysis. A comprehensive analysis of the collected data revealed that high serum CA 19-9 levels before treatment were significantly associated with poor OS (HR: 1.58, 95% CI: 1.36-1.83, P<0.001), DFS (HR: 1.71, 95% CI: 1.38-2.13, P<0.001), PFS (HR: 1.30,95%CI:0.93-1.82, P = 0.121) and RFS (HR: 1.43, 95% CI: 1.11-1.83, P = 0.006). This association between high pretreatment serum CA 19-9 levels and poor survival held true across different geographical regions, analysis types, methods used for HR determination, sample size, and treatment methods. CONCLUSIONS: The results of this study indicate that pretreatment serum CA 19-9 level can be used as a prognostic indicator for patients with CRC.

A rare case of eight multiple primary malignant neoplasms in a female patient: A case report and review of the literature.

In recent years, the number of patients exhibiting multiple primary malignant neoplasms has been increasing. The current study reports the rare case of a 61-year-old Chinese female with eight histopathologically confirmed primary malignant neoplasms; to the best of our knowledge, such a high number of malignant neoplasms in one patient has not previously been described in the English literature. The tumors originated from the colon (five separate tumors over 32 years), the endometrium, the breast and the small intestine. It is important to diagnose multiple primary malignant neoplasms as such patients generally exhibit a more favorable prognosis compared with metastatic carcinoma patients. In addition, prolonged follow-up after surgery should be considered.

Expression of the co-signaling molecules CD40-CD40L and their growth inhibitory effect on pancreatic cancer in vitro.

We investigated the expression of the co-signalling molecule CD40 in pancreatic cancer and the growth inhibitory effect of the recombinant soluble human CD40 ligand (rshCD40L) in pancreatic cancer cell lines. Twenty-six cases of pancreatic cancer tissues and corresponding paratumoral normal tissues were immunohistochemically analyzed for CD40 expression. The association of CD40 expression with clinicopathological parameters, including clinical stage, pathological grade, invasion and metastasis, were statistically analyzed. The serum sCD40 levels in pancreatic cancer patients were examined by ELISA. The expression of CD40 in the pancreatic cancer cell lines Panc-1, Aspc-1 and Miapaca-2 was examined by RT-PCR and flow cytometry. The growth inhibitory activity of rshCD40L on pancreatic cancer cell lines was determined by MTT assay. Tumor cell apoptosis was detected by TUNEL and Annexin V/PI double staining method. CD40 was positive both on the membrane and in the cytoplasm of tumor cells, 69.2% (18/26) of the cases were positive for CD40. CD40 expression was significantly higher in pancreatic cancer tissues compared to adjacent normal tissues (P<0.05). High CD40 expression was associated with TNM stage and lymph node metastasis (both P<0.05). Patients with pancreatic cancer have higher serum sCD40L levels (3.53 ± 0.70 ng/ml) compared to healthy subjects (1.81 ± 0.48 ng/ml, P<0.05). rshCD40L significantly inhibited the proliferation of the pancreatic cancer cell lines and induced apoptosis in these cell lines. The co-signaling molecule CD40 is highly expressed in pancreatic cancer tissues and cell lines and rshCD40L is a potential tool for antitumor therapies.