Osteosarcoma is the most common primary sarcoma of the bone. Lung osteosarcoma metastases at diagnosis have a significantly poor prognosis, even when surgery plus chemotherapy are performed. Our goal was to analyze clinical and sarcoma characteristics that could help identify factors related to an increased rate of lung metastasis and to identify different modes of treatment and its correlation with survival. The Surveillance, Epidemiology, and End Results (SEER) database was used to identify all osteosarcoma patients diagnosed from 2010 to 2015. Patient characteristics such as age, sex, ethnicity, marital status, tumor location, histologic grade, surgery, chemotherapy, radiation therapy, SEER cause-specific death classification, survival, and lung metastasis were collected. These factors were analyzed using Univariate and multivariate regression models in survival analyses. A total of 1057 osteosarcoma patients diagnosed from 2010 to 2015 were included, of which 176 were patients with lung metastasis. Substantial disparities in the rate of lung metastasis existed when osteosarcoma patients were stratified according to tumor location (P = 0.0002) and tumor size (P < .001). Using a Multivariate Cox regression model, being older than 30 years (vs. younger than 30, HR = 2.171, 95% CI = 1.623-2.905, P < .0001), having a tumor >5-10 cm (vs. 10 cm (vs. <5 cm, HR = 3.610, 95% CI = 2.066-6.310, P < .0001) were related to an increased HR for all-cause death. The HR decreased in patients with surgery (vs. no surgery, HR = 0.189, 95% CI = 0.138-0.260, P < 0.0001) and osteosarcoma. As for osteosarcoma patients with lung metastases, Multivariate Cox regressions revealed that an increased HR was associated with being older than 30 years (vs. younger than 30 years, HR = 2.142, 95% CI = 1.273-3.605, P = .0041) and married (vs. no marriage, HR = 2.418, 95% CI = 1.400-4.176, P= .0015), while a decreased HR was related to having had surgery (vs. no surgery, HR = 0.282, 95% CI = 0.171-0.464, P < .0001) and chemotherapy (vs. no chemotherapy, HR = 0.107, 95% CI = 0.050-0.229, P < .0001). Advanced age (older than 30 years) and large tumors were related to a higher risk of lung metastases in osteosarcoma patients. Therefore, patients who were diagnosed at advanced age or had large tumors should receive comprehensive chest CT scans. Surgery and chemotherapy can significantly improve the survival of metastatic patients, while radiotherapy did not improve survival in these patients.

Background: Pancreatic cancer is one of the most lethal cancers worldwide. The aim of this study was to determine the expression pattern, clinical significance, and biological functions of Wnt5a in pancreatic cancer. Methods: Immunohistochemistry was performed to examine Wnt5a expression in 134 surgically resected pancreatic adenocarcinoma and adjacent normal pancreatic tissues. Associations of Wnt5a expression with clinicopathological factors and cancer-specific survival were analyzed. The effects of Wnt5a overexpression or silencing on the invasiveness and epithelial-to-mesenchymal transition (EMT) of pancreatic cancer cells were studied. Silencing of β-catenin by small interfering RNA was done to determine its role in the Wnt5a-mediated tumor phenotype. Results: The percentage of Wnt5a positive expression showed a bell-shaped pattern in pancreatic cancer tissues, peaking in well-differentiated carcinomas. The median cancer-specific survival was comparable between patients with positive versus negative expression of Wnt5a. Overexpression of Wnt5a promoted the migration and invasion of pancreatic cancer cells, whereas Wnt5a depletion had an inhibitory effect. In an orthotopic pancreatic cancer mouse model, Wnt5a overexpression resulted in increased invasiveness and metastasis, coupled with induction of EMT in tumor cells. Treatment with recombinant Wnt5a elevated the nuclear β-catenin level in pancreatic cancer cells, without altering the Ror2 expression. Targeted reduction of β-catenin antagonized exogenous Wnt5a-induced EMT and invasiveness in pancreatic cancer cells. Conclusion: Upregulation of Wnt5a promotes EMT and metastasis in pancreatic cancer models, which involves activation of β-catenin-dependent canonical Wnt signaling. These findings warrant further investigation of the clinical relevance of Wnt5 upregulation in pancreatic cancer.

Perineural invasion and immunosuppressive tumor microenvironment are the distinct features of pancreatic ductal adenocarcinoma (PDAC). Heterogeneous myeloid-derived suppressor cells (MDSCs) are potent suppressors of antitumor immunity, posing obstacles for cancer immunotherapy. Increasing evidences have demonstrated the accumulation of MDSCs in PDAC patients. However, the role of MDSCs in perineural invasion of PDAC and the existence of novel MDSC subsets during PDAC remain unclear. This study found that lymphocytic perineural cuffs were frequently present in chronic pancreatitis (CP) tissues and adjacent non-neoplastic pancreatic tissues (ANPTs), but not in PDAC with perineural invasion. Meanwhile, we found that neutrophil-like MDSCs (nMDSCs), but not monocyte-like MDSCs (mMDSCs), were significantly increased in PBMCs and tumor tissues of PDAC patients. Further observation identified two distinct subsets of nMDSCs, CD13^hi and CD13^low nMDSCs in PDAC patients, which have not been reported previously. Despite a similar morphology, CD13^hi nMDSCs expressed higher levels of CD11b, CD33, CD16 and arginase 1 but lower levels of CD66b than CD13^low nMDSCs. Importantly, CD13^hi MDSCs, compared with CD13^low nMDSCs, more effectively suppressed alloreactive T cell responses via an arginase-1-related mechanism. After tumor resection, the circulating CD13^hi nMDSCs were decreased markedly. PDAC patients with more CD13^hi nMDSCs had a shorter overall survival than those with less CD13^hi nMDSCs. To conclude, we identified two novel MDSC subsets with different characteristics and functions in PDAC, demonstrated the association of the two MDSC subsets with cancer progression, and explored their roles in perineural invasion and immune escape of PDAC.

Toll-like receptor (TLR) signaling plays major roles in innate immune response in macrophages. Melatonin regulates TLR3- and TLR4-mediated innate immune responses in macrophages. However, it remains unknown whether melatonin regulates TLR9-mediated innate immune responses in macrophages. Here we demonstrated that melatonin suppressed TLR9 ligand-induced proinflammatory cytokines mRNA and protein production in peritoneal macrophages without interrupting the viability of peritoneal macrophages. Using a melatonin membrane receptors MT1/MT2 antagonist luzindole, we found that MT1 and MT2 were dispensable for melatonin’s inhibitory effects on TLR9-mediated proinflammatory cytokines production, even though melatonin upregulated mRNA expression of MT1 and MT2 in macrophages. Furthermore, melatonin did not affect mRNA expressions of TLR9 and MyD88 but attenuated TLR9 ligand-induced ERK1/2 and AKT phosphorylation without affecting p38 and NF-κB p65 phosphorylation. Also, melatonin inhibited TLR9-mediated proinflammatory cytokines productionin vivo. Taken together, our results demonstrate that melatonin suppresses TLR9-triggered proinflammatory cytokines production in macrophages via melatonin membrane receptor-independent manners and probably through inhibiting ERK1/2 and AKT activation, which further elucidates the roles of melatonin in regulating TLR-mediated innate immune responses in macrophages.

To investigate the influence of region of interest (ROI) size on tumor apparent diffusion coefficient (ADC) measurements in pancreatic cancer. The study population consisted of 64 patients with pathologically proved pancreatic ductal adenocarcinomas (PDACs), who underwent preoperative magnetic resonance imaging (MRI) examinations including diffusion-weighted imaging (DWI). The tumor ADCs were measured by two independent readers using six round ROIs with sizes ranging from 20 to 214 mm2 (9 to 97 pixels) in both the six separate measurements. The intra- and inter-observer variabilities were analyzed by using the coefficient of variance (CV), the interclass correlation coefficient (ICC) and Bland-Altman analysis. The mean ADCs measured with the 6 different-sized ROIs were compared using one-way repeated analysis of variance. The sample sizes were calculated by using 80% power and a 5% significance level to detect 10 to 25% changes in ADC measurements. The largest ROI (ROI214) yielded the best intra-observer repeatability (CV, 6.3%; ICC, 0.93) and inter-observer reproducibility (CV, 10.1%; ICC, 0.84). The mean differences in ADC measurements ± limits of agreement between the two readers were (0.06 ± 0.47) × 10−3 mm2 for ROI20, (0.08 ± 0.46) × 10−3 mm2 for ROI46, (0.05 ± 0.37) × 10−3 mm2 for ROI82, (0.07 ± 0.42) × 10−3 mm2 for ROI115, (0.05 ± 0.43) × 10−3 mm2 for ROI152 and (−0.02 ± 0.29) × 10−3 mm2 for ROI214. ROI size had a considerable influence on the ADC measurements of PDACs.

Clinical and Molecular Hepatology