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Supplementary Figure 3. The mRNA expression of CTTN in rhVEGF-C-treated TE-1 cells. TE-1 cells were treated with a various doses of rhVEGF-C (0, 25, 50, 100 and 200 ng/mL) for 24 hours and isolated total RNA to measure the expression level of CTTN by RT-PCR. GAPDH was used as an internal control.

Supplementary Figure 1. The mRNA expression of Nrp1 and Nrp2 in ESCC cell lines. RT-PCR was showed the expression level of Nrp1 and Nrp2 in TE1, TE12, TE3, TE7, and TE8 cells. GAPDH was used as an internal control.

Supplementary Information

Supplementary Figure 4. The mRNA expression of miR-326 in TE-8 cells with knockdown of VEGFR-3. TE-8 cells were transiently transfected shVEGFR-3#1, #2 or shCtrl for 48 hours and isolated total RNA to assay. A, qRT-PCR was used to quantify the expression level of miR-326 in those indicated cells. B, RT-PCR was showed the expression level of VEGFR-3 in those indicated cells. GAPDH was used as an internal control. The quantities values were evaluated as percentage compared with TE-8/shCtrl cells. The columns are the mean values from three independent experiments. Bars indicate means {plus minus} SE.

Supplementary Figure 5. The mRNA expression of Dicer between TE-1/VEGF-C and TE-1/vector cells. TE-1 cells were stably expressed VEGF-C or vector control and isolated total RNA to assay. qRT-PCR was used to quantify the expression level of Dicer, and the quantities values were evaluated as percentage compared with TE-1/vector cells. The columns are the mean values from three independent experiments. Bars indicate means {plus minus} SE.

Supplementary Figure 2. The phosphorylated VEGFR-3 in TE-1 cells with VEGF-C and VEGF-A treatment. TE-1 cells were starved with serum-free medium for 16 hours and treated with 100 and 200 ng/mL VEGF-C as well as 200 ng/mL VEGF-A for 10 minutes, following examination of phosphorylated VEGFR-3 (p-VEGFR-3, Y1063/1068) expression by Western blot. β-actin was used as an internal control.

Protein disulfide isomerases a4 (Pdia4) is known to be involved in cancer development. Our previous publication showed that Pdia4 positively promotes cancer development via its inhibition of procaspase-dependent apoptosis in cancer cells. However, nothing is known about its role in the cancer microenvironment.Here, we first found that Pdia4 expression in lung cancer was negatively correlated with patient survival. Next, we investigated the impact of host Pdia4 in stromal cells during cancer development. We showed that Pdia4 was expressed at a low level in stromal cells, and this expression was up-regulated akin to its expression in cancer cells. This up-regulation was stimulated by tumour cell-derived stimuli. Genetics studies in tumour-bearing wild-type and Pdia4These findings identify Pdia4 as a possible target for intervention in cancer stroma, suggesting that targeting Pdia4 in cancer stroma is a promising anti-cancer approach.

Controlling the behavior of mesenchymal stem cells (MSCs) through topographic patterns is an effective approach for stem cell studies. We, herein, reported a facile method to create a dopamine (DA) pattern on poly(dimethylsiloxane) (PDMS). The topography of micropatterned DA was produced on PDMS after plasma treatment. The grid-topographic-patterned surface of PDMS-DA (PDMS-DA-P) was measured for adhesion force and Young's modulus by atomic force microscopy. The surface of PDMS-DA-P demonstrated less stiff and more elastic characteristics compared to either nonpatterned PDMS-DA or PDMS. The PDMS-DA-P evidently enhanced the differentiation of MSCs into various tissue cells, including nerve, vessel, bone, and fat. We further designed comprehensive experiments to investigate adhesion, proliferation, and differentiation of MSCs in response to PDMS-DA-P and showed that the DA-patterned surface had good biocompatibility and did not activate macrophages or platelets in vitro and had low foreign body reaction in vivo. Besides, it protected MSCs from apoptosis as well as excessive reactive oxygen species (ROS) generation. Particularly, the patterned surface enhanced the differentiation capacity of MSCs toward neural and endothelial cells. The stromal cell-derived factor-1α/CXantiCR4 pathway may be involved in mediating the self-recruitment and promoting the differentiation of MSCs. These findings support the potential application of PDMS-DA-P in either cell treatment or tissue repair.

Epidermal growth factor receptor (EGFR), a critical factor promotes lung cancer cell proliferation and survival. Knockdown of EGFR expression thus promise beating lung cancer clinically. Functionalized gold nanoparticles may serve an effective vehicle carrying theranostic bio-active materials. Herein, physically gold nanoparticles were fabricated with biocompatible collagen to improve siRNA loading capacity carrying EGFR siRNA to treat lung cancer. Physic-chemical properties were comprehensively characterized for the collagen gold nanoparticle (C–Au), and with EGFR siRNA conjugation, C–Au-EGFRsi namely. Issues of biocompatibility were addressed. Interestingly, C–Au appeared more biocompatible to normal airway epithelial cells (BEAS-2B) than to cancer cells (A549) in terms of ROS production, cell cycle behavior, and cell growth influence. The C–Au demonstrated comparable or even more efficient, compared with lipofetamine, in carrying siRNA to knockdown EGFR of A549 cells. Endocytosis mediated cell entry for the collagen gold nanoparticles, and endosome-lysosomal pathway involved transporting and metabolizing these nanoparticles. In xenograft mice model, substantial tumor suppression effects were observed treating with C–Au-EGFRsi, in which tumor weight reduced 30% for lipofetamine carrier, and down to 70% for C–Au carrier. Particularly, overall survival rate was improved for both treatment groups with lipofetamine and C–Au carrier, respectively.

Combination of cancer immunotherapies and nanotechnology has shown to increase therapeutic efficiency and efficacy with less safety concerns. For an effective vaccine, activation and functional maturation of dendritic cells is a key step for the subsequent adaptive immune responses against cancer. Owing to superior biocompatibility, tunable physicochemical properties, low toxicity in immune cells and ease of functionalization by modifying surface with various biomolecules, gold nanoparticles (AuNP) has been exploded for the development of better dendritic cells (DCs)-based vaccines. AuNPs with biopolymer such as collagen has been revealed to increase biological performance. In the present study, a novel nanocarrier comprising of Au nanoparticles fabricated with collagen biomolecule was used to carry bioactive molecules called stromal-derived factor-1α (SDF-1α), which is known to enhance DCs survival, maturation, and migration, Dynamic light scattering (DLS) analysis showed the particle size of Au-Col-SDF-1α (Au-Col-SDF-1) was 308 ± 3.3 nm. Both UV-Vis absorption and FTIR spectrum analysis demonstrated that SDF-1 successfully conjugated onto Au-Col. Cellular uptake analysis indicated that DCs exhibited an excellent uptake capacity when SDF-1α was carried by Au-Col nanocarrier. In addition, MTT assay and cell cycle analysis revealed that Au-Col-SDF-1 could facilitate viability and proliferation of DCs. Furthermore, Au-Col-SDF-1 could promote DCs migration and upregulate matrix metalloproteinase (MMP) enzymatic activity in DCs as well as enhance the ability of DCs in inducing T cell cytokines production. Moreover, Au-Col-SDF-1 could stimulate DCs to express CXCR4 then secret more SDF-1α to further influence the surrounding DCs. In conclusion, our findings provide a new application by promoting DCs cell viability, migration and ability to enhance T cell cytokine production for a better DC‐based cancer vaccine using a novel AuNP-Col nanocarrier that was bounded with SDF-1α (Au-Col-SDF-1α)

The engineering of vascular regeneration still involves barriers that need to be conquered. In the current study, a novel nanocomposite comprising of fibronectin (denoted as FN) and a small amount of silver nanoparticles (AgNP, ~15.1, ~30.2 or ~75.5 ppm) was developed and its biological function and biocompatibility in Wharton’s jelly-derived mesenchymal stem cells (MSCs) and rat models was investigated. The surface morphology as well as chemical composition for pure FN and the FN-AgNP nanocomposites incorporating various amounts of AgNP were firstly characterized by atomic force microscopy (AFM), UV-Visible spectroscopy (UV-Vis), and Fourier-transform infrared spectroscopy (FTIR). Among the nanocomposites, FN-AgNP with 30.2 ppm silver nanoparticles demonstrated the best biocompatibility as assessed through intracellular ROS production, proliferation of MSCs, and monocytes activation. The expression levels of pro-inflammatory cytokines, TNF-α, IL-1β, and IL-6, were also examined. FN-AgNP 30.2 ppm significantly inhibited pro-inflammatory cytokine expression compared to other materials, indicating superior performance of anti-immune response. Mechanistically, FN-AgNP 30.2 ppm significantly induced greater expression of vascular endothelial growth factor (VEGF) and stromal-cell derived factor-1 alpha (SDF-1α) and promoted the migration of MSCs through matrix metalloproteinase (MMP) signaling pathway. Besides, in vitro and in vivo studies indicated that FN-AgNP 30.2 ppm stimulated greater protein expressions of CD31 and von Willebrand Factor (vWF) as well as facilitated better endothelialization capacity than other materials. Furthermore, the histological tissue examination revealed the lowest capsule formation and collagen deposition in rat subcutaneous implantation of FN-AgNP 30.2 ppm. In conclusion, FN-AgNP nanocomposites may facilitate the migration and proliferation of MSCs, induce endothelial cell differentiation, and attenuate immune response. These finding also suggests that FN-AgNP may be a potential anti-inflammatory surface modification strategy for vascular biomaterials.

Incense burning is a popular cultural and religious practice, but whether exposure to incense smoke has effects on lung function is unclear. We investigated association between lung function and incense burning exposure and other household exposures in adolescents who participated in a mass asthma-screening program. Information on asthmatic status and associated factors was obtained from parent completed questionnaires and student-completed video questionnaires. Approximately 10% of students received lung-function examinations. Valid lung-function data of 5010 students aged 14–16 years in northern Taiwan were analyzed. Forced vital capacity (FVC) and forced expiratory flow in one second (FEV1) were compared by incense burning status and other types of exposures for adolescents. Overall, 70.6% of students were exposed to incense smoke at home. The mean FVC and FEV1 measures were lower among adolescents with daily exposure to incense burning than those without such exposure (p

The regulating storage tank plays an important role in water pollution. However, a complete system of evaluation methods for site selection of regulating storage tank is still missing. To solve the problem, first, the influence factors are established through the interpretative structural modeling, and the corresponding membership functions for each factor are established. Second, a determining method for evaluation index weight is adopted based on triangle fuzzy number and analytic hierarchy process method. Then, a fuzzy comprehensive evaluation is constructed and made for site selection of the regulating storage tanks in a sponge city, where the regulating storage tanks are located in the “ehu park and binhe road junction” of Pingxiang city, China. Finally, the results of fuzzy comprehensive evaluation for site selection of the regulating storage tanks are obtained, and the proposed approach is proved to be effective and reliable.

Stereotactic body radiation therapy (SBRT) plays an important role in early stage non-small cell lung cancer. Tumor growth before radiotherapy planning (RTP) or during SBRT has been reported in lung cancer patients; however, little is known of growth during the period in-between (i.e. after RTP but before SBRT). An 83-year-old man referred to our hospital and diagnosed with medically inoperable non-small cell lung cancer was noted to have significant tumor progression on day 1 of cone beam computed tomography just before the planned SBRT delivery. Because of uncertainty of the underlying etiology and unfamiliarity with this phenomenon, we made a clinical decision to arrange re-simulation and revise our treatment to conventional fractionated radiotherapy (CFRT). After an initial response, distant metastases occurred eight months after CFRT. The patient received best supportive care and was under hospice care at the last follow-up (27 months after CFRT). We report a case with significant tumor growth just before planned SBRT. Optimal management in this scenario requires further investigation.

Gliomas are the most common primary brain malignancies, with different degrees of aggressiveness, variable prognosis and various heterogeneous histologic sub-regions, i.e., peritumoral edematous/invaded tissue, necrotic core, active and non-enhancing core. This intrinsic heterogeneity is also portrayed in their radio-phenotype, as their sub-regions are depicted by varying intensity profiles disseminated across multi-parametric magnetic resonance imaging (mpMRI) scans, reflecting varying biological properties. Their heterogeneous shape, extent, and location are some of the factors that make these tumors difficult to resect, and in some cases inoperable. The amount of resected tumor is a factor also considered in longitudinal scans, when evaluating the apparent tumor for potential diagnosis of progression. Furthermore, there is mounting evidence that accurate segmentation of the various tumor sub-regions can offer the basis for quantitative image analysis towards prediction of patient overall survival. This study assesses the state-of-the-art machine learning (ML) methods used for brain tumor image analysis in mpMRI scans, during the last seven instances of the International Brain Tumor Segmentation (BraTS) challenge, i.e., 2012-2018. Specifically, we focus on i) evaluating segmentations of the various glioma sub-regions in pre-operative mpMRI scans, ii) assessing potential tumor progression by virtue of longitudinal growth of tumor sub-regions, beyond use of the RECIST/RANO criteria, and iii) predicting the overall survival from pre-operative mpMRI scans of patients that underwent gross total resection. Finally, we investigate the challenge of identifying the best ML algorithms for each of these tasks, considering that apart from being diverse on each instance of the challenge, the multi-institutional mpMRI BraTS dataset has also been a continuously evolving/growing dataset., Comment: The International Multimodal Brain Tumor Segmentation (BraTS) Challenge

In Taiwan, lung cancer remains one of the deadliest cancers. Survival of lung cancer patients remains low, ranging from 6% to 18%. Studies have shown that Chinese herbal medicine (CHM) can be used to induce cell apoptosis and exhibit anti-inflammatoryanti-inflammatory activities in cancer cells.This study aimed to investigate the frequencies and patterns of CHM treatment for lung cancer patients and the effect of CHM on their survival probability in Taiwan.We identified 6939 lung cancer patients (ICD-9-CM: 162). We allocated 264 CHM users and 528 CHM-non users, matched for age, gender, duration, and regular treatment. Chi-square test, conditional multivariable logistic regression, Kaplan-Meier method, and the log-rank test were used in this study.The CHM group was characterized by a longer follow up time and more cases of hyperlipidemia and liver cirrhosis. This group exhibited a lower mortality hazard ratio (0.48, 95% confidence interval [0.39-0.61], p0.001), after adjusting for comorbidities. The trend was also observed that the cumulative survival probability was higher in CHM than in non-CHM users (p0.0001, log rank test). Analysis of their CHM prescription pattern revealed that Bu-Zhong-Yi-Qi-Tang (BZYQT), Xiang-Sha-Liu-Jun-Zi-Tang (XSLJZT), and Bai-He-Gu-Jin-Tang (BHGJT); and Bei-Mu (BM), Xing-Ren (XR) and Ge-Gen (GG) were found to be the top three formulas and herbs, respectively. Among them, BM was the core CHM of the major cluster, and Jie-Geng (JG) and Mai-Men-Dong-Tang (MMDT) were important CHMs by CHM network analysis.The use of CHM as an adjunctive therapy may reduce the mortality hazard ratio of lung cancer patients. The investigation of their comprehensive CHM prescription patterns might be useful in future large-scale, randomized clinical investigations of agent effectiveness, safety, and potential interactions with conventional treatments for lung cancer patients.

Esophageal cancer is an aggressive human malignancy with increasing incidence in the developed world. VEGF-C makes crucial contributions to esophageal cancer progression that are not well understood. Here, we report the discovery of regulatory relationship in esophageal cancers between the expression of VEGF-C and cortactin (CTTN), a regulator of the cortical actin cytoskeleton. Upregulation of CTTN expression by VEGF-C enhanced the invasive properties of esophageal squamous cell carcinoma in vitro and tumor metastasis in vivo. Mechanistic investigations showed that VEGF-C increased CTTN expression by downregulating Dicer-mediated maturation of miR326, thereby relieving the suppressive effect of miR326 on CTTN expression. Clinically, expression of Dicer and miR326 correlated with poor prognosis in patients with esophageal cancer. Our findings offer insights into how VEGF-C enhances the robust invasive and metastatic properties of esophageal cancer, which has potential implications for the development of new biomarkers or therapies in this setting. Cancer Res; 74(21); 6280–90. ©2014 AACR.

Vascular endothelial growth factor-C (VEGF-C) plays an important role during cancer progression and metastasis through activation of VEGF receptors. However, the role of VEGF-C in esophageal squamous cell carcinoma (ESCC) remains unclear. The expression of VEGF-C in advanced stages of esophageal cancer was examined by immunohistochemistry and its expression was correlated with the protein level of cortactin (CTTN) by Western blot. Knockdown and overexpression of the CTTN protein were respectively performed to investigate the effects on VEGF-C-enhanced ESCC migration/invasion by in vitro transwell assay, cell tracing assay, and tumor growth/experimental metastasis in animal models. The expression of VEGF-C was positively correlated with tumor status and poor clinical prognosis in patient with esophageal cancer. VEGF-C-upregulated CTTN expression contributed the migration/invasive abilities of ESCC cell lines through Src-mediated downregulation of miR-326. Moreover, knockdown of CTTN expression significantly abolished VEGF-C-induced tumor growth and experimental lung metastasis in vivo. Upregulation of CTTN is critical for VEGF-C-mediated tumor growth and metastasis of ESCC. These finding suggest that VEGF-C upregulated CTTN expression through Src-mediated downregulation of miR-326. CTTN may be a crucial mediator of VEGF-C-involved ESCC metastasis, which provides a potential target for diagnosis and individualized treatment in clinical practice.

Accumulating evidence is revealing an important role of microRNA (miRNA) in tumor progression and chemotherapeutic resistance. Dicer is a cytoplasmic endoribonuclease type III crucial for production of mature miRNAs. The aberrant expression of Dicer has also been reportedly associated with clinical aggressiveness, prognosis, and patient survival in various cancer types. However, the molecular mechanisms of Dicer in acquired gefitinib resistance are still not clear.In this study, we analyzed the protein level of Dicer between gefitinib-sensitive (PC9) and gefitinib-resistant (PC9/GR) non-small-cell lung cancer (NSCLC) cell lines by Western blot analysis. Silence and overexpression of the Dicer were performed to investigate the effects on gefitinib sensitivity, as assessed by (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay and sub-G1 assay of flow cytometry. To further explore the mechanism of chemoresistance, we examined whether Dicer knockdown led to modulating specific miRNAs and its miRNA target genes.Dicer expression was significantly increased in PC9/GR compared with PC9 cells. Knockdown of Dicer restores gefitinib sensitivity in resistant cells, and overexpression of Dicer enhances resistance to gefitinib in sensitive cells. Silencing of Dicer induces sensitivity to gefitinib in NSCLC cells through the downregulation of miR-30b/c and miR-221/222 to increase the protein level of caspase-3, resulting in an increase in gefitinib-induced apoptosis.Dicer contributes to the resistance to gefitinib in lung cancer. These results indicate that Dicer may be a target for diagnosis and therapy of patients with resistance to gefitinib.

Aims and objectives To develop a Regular Exercise Belief Questionnaire and test its psychometric properties for patients with chronic obstructive pulmonary disease. Background Regular exercise has been shown to significantly improve physical capacity and the quality of life of patients with chronic obstructive pulmonary disease. However, their adherence to long-term exercise is low. To develop an effective strategy for promoting good exercise behaviours, it is important to have a validated instrument to evaluate factors related to engaging in exercise. Design and methods A cross-sectional design was used for the study. Construction of the Regular Exercise Belief Questionnaire was based on the Theory of Planned Behaviour Reliability and validity were assessed using a sample of 136 male patients with chronic obstructive pulmonary disease. The construct validity of the questionnaire was confirmed through exploratory factor analysis and known group technique. Results Exploratory factor analysis resulted in an eight-factor solution that explained 70·4% of the total variance. The internal consistency of the Regular Exercise Belief Questionnaire was 0·83–0·93. The Regular Exercise Belief Questionnaire was preliminarily found to be reliable and exhibited satisfactory validity for patients with chronic obstructive pulmonary disease. Conclusion The Regular Exercise Belief Questionnaire is the first theory-based measure of exercise beliefs among patients with chronic obstructive pulmonary disease. The questionnaire provides an effective method to examine behaviour beliefs, normative beliefs and control beliefs about regular exercise. Clinical relevance The measure can be used to periodically evaluate the exercise beliefs in clinics and to examine the effectiveness of exercise programmes in patients with chronic obstructive pulmonary disease. The result of the evaluation could also apply to identify strategies related to promoting exercise behaviours.

Objectives To explore the beliefs regarding regular exercise among patients with chronic obstructive pulmonary disease (COPD). Background Low adherence to exercise has been observed in patients with COPD. It is important to identify factors regarding exercise from the patients' viewpoint. Methods Thirty-one patients were recruited from a medical center in Taiwan. Semi-structured, in-depth, one-on-one interviews were conducted to collect data. The narratives of the interviews were analyzed via content analysis. Results The majority of the participants affirmed the benefits of regular exercise; however, concerns about personal comfort and safety affected their actual exercise behavior. Five normative references were found to support exercise behavior, and several exercise promoters were identified. Conclusion This study provides an understanding of exercise beliefs of Taiwanese patients with COPD stage II–IV and suggests several ideas for their exercise maintenance. There is a need to provide individualized exercise guides and reinforced programs for patients with chronic obstructive pulmonary disease.

Objective: To investigate lung function associated with asthma and body mass index (BMI) among adolescents at 96 northern Taiwan junior high schools participating in an asthma screening program. Methods: The questionnaires and lung function test results measured for 3669 boys and 3523 girls were included in this study for data analysis. Measures of forced vital capacity (FVC), forced expiratory volume in 1 second (FEV1) and FEV1/FVC ratio were compared by sex, asthma status and BMI. Results: Overall mean FVC levels were similar between students with and without asthma, 3.71 L vs. 3.71 L for boys (p = 0.991) and 2.79 vs. 2.78 for girls (p = 0.517). The overall mean FEV1 levels were also similar between girls with and without asthma. Asthmatic boys had lower FEV1 than non-asthmatic boys. Mean FEV1/FVC was significantly lower in students with asthma than those without asthma. Mean FVC and FEV1 increased with BMI in both sexes. A lower mean FEV1/FVC was observed among students with asthma and high BMI, and was more pronounced in boys than in girls. Multivariable regression analysis also showed that FEV1/FVC ratios were negatively associated with asthma and high BMI, stronger in boys than in girls for asthma (β = −2.176 (standard errors (SE) = 0.268) vs. −1.085 (SE = 0.258) and for BMI (β = −0.309 (SE = 0.025) vs. −0.218 (SE = 0.029)). Conclusion: This northern Taiwan study suggests that FEV1/FVC is negatively associated with asthma and high BMI in adolescents, stronger for boys than for girls.

Therapy with epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (EGFR-TKIs, such as gefitinib or erlotinib) significantly prolongs survival time for patients with tumors harboring an activated mutation on EGFR; however, up to 40% of lung cancer patients exhibit acquired resistance to EGFR-TKIs with an unknown mechanism. FOXO3a, a transcription factor of the forkhead family, triggers apoptosis, but the mechanistic details involved in EGFR-TKI resistance and cancer stemness remain largely unclear. Here, we observed that a high level of FOXO3a was correlated with EGFR mutation-independent EGFR-TKI sensitivity, the suppression of cancer stemness, and better progression-free survival in lung cancer patients. The suppression of FOXO3a obviously increased gefitinib resistance and enhanced the stem-like properties of lung cancer cells; consistent overexpression of FOXO3a in gefitinib-resistant lung cancer cells reduced these effects. Moreover, we identified that miR-155 targeted the 3′UTR of FOXO3a and was transcriptionally regulated by NF-κB, leading to repressed FOXO3a expression and increased gefitinib resistance, as well as enhanced cancer stemness of lung cancer in vitro and in vivo. Our findings indicate that FOXO3a is a significant factor in EGFR mutation-independent gefitinib resistance and the stemness of lung cancer, and suggest that targeting the NF-κB/miR-155/FOXO3a pathway has potential therapeutic value in lung cancer with the acquisition of resistance to EGFR-TKIs.

Resveratrol, a phytochemical found in various plants and Chinese herbs, is associated with multiple tumor-suppressing activities, has been tested in clinical trials. However, the molecular mechanisms involved in resveratrol-mediated tumor suppressing activities are not yet completely defined. Here, we showed that treatment with resveratrol inhibited cell mobility through induction of the mesenchymal-epithelial transition (MET) in lung cancer cells. We also found that downregulation of FOXC2 (forkhead box C2) is critical for resveratrol-mediated suppression of tumor metastasis in an in vitro and in vivo models. We also identified a signal cascade, namely, resveratrol-∣miRNA-520h-∣PP2A/C-∣Akt → NF-κB → FOXC2, in which resveratrol inhibited the expression of FOXC2 through regulation of miRNA-520h-mediated signal cascade. This study identified a new miRNA-520h-related signal cascade involved in resveratrol-mediated tumor suppression activity and provide the clinical significances of miR-520h, PP2A/C and FOXC2 in lung cancer patients. Our results indicated a functional link between resveratrol-mediated miRNA-520h regulation and tumor suppressing ability, and provide a new insight into the role of resveratrol-induced molecular and epigenetic regulations in tumor suppression.

Ganoderma tsugae(GT) is a traditional Chinese medicine that exhibits significant antitumor activities against many types of cancer. This study investigated the molecular mechanism by which GT suppresses the growth of doxorubicin-resistant lung adenocarcinoma H23/0.3 cells. Our results reveal that GT inhibits the viability of H23/0.3 cellsin vitroandin vivoand sensitizes the growth suppression effect of doxorubicin on H23/0.3 cells. The data also show that GT induces S phase arrest by interfering with the protein expression of cyclin A, cyclin E, CDK2, and CDC25A. Furthermore, GT induces cellular apoptosis via induction of a mitochondria/caspase pathway. In addition, we also demonstrate that the suppression of cell proliferation by GT is through down-regulation of the PI3K/Akt signaling pathway. In conclusion, this study suggests that GT may be a useful adjuvant therapeutic agent in the treatment of lung cancer.

Background and Purpose— Stroke is a known cerebrovascular complication in lung cancer patients; however, whether lung cancer patients are at elevated risk of developing stroke relative to the noncancer population remains unclear. Methods— The present study used population-based claims data from the Taiwan National Health Insurance, which identified 52 089 patients with an initial diagnosis of lung cancer between 1999 and 2007, and 104 178 matched noncancer subjects from all insured subjects age 20 years and older. Subsequent occurrence of stroke was measured until 2008, and the association between lung cancer and the hazard of developing stroke was estimated using Cox proportional hazard models. Results— The incidence of stroke was 1.5 times higher (25.9 versus 17.4 per 1000 person–years) in the lung cancer group compared with the comparison group. The multivariate-adjusted hazard ratio (HR) comparing lung cancer patients with the noncancer group was 1.47 (95% CI, 1.39–1.56) for stroke, 1.78 (95% CI, 1.54–2.05) for hemorrhagic stroke, and 1.43 (95% CI, 1.34–1.51) for ischemic stroke. The risk of stroke fell over time, decreasing after 1 year of follow-up for men and after 2 years of follow-up for women. Within the first year of follow-up, the risk of stroke peaked during the first 3 months for men and within 4 to 6 months for women. Conclusions— Lung cancer is associated with increased risk of subsequent stroke within 1 year after diagnosis for men and 2 years after diagnosis for women.

NOD2 of the NLRs and TLR4 of the TLRs are major pattern-recognition receptors, which sense different microbial pathogens and have important roles in innate immunity. Herein, we investigated the roles of NOD2 in TLR4-mediated signalling and gene regulation in RAW264.7 macrophages. We found that MDP (a NOD2 ligand) increased LPS-induced expressions of TNF-α, IL-1β, IL-6, iNOS and COX-2. MDP did not affect LPS-induced activation of MAPKs or IKK, while it potentiated LPS-induced NF-κB activation. Meanwhile TLR4 activation increased NOD2 mRNA expression, and upregulated NOD2 upon MDP treatment is a positive regulator of TLR4-mediated signalling. Intriguingly we found that NOD2 silencing led to increases in LPS-induced signal transduction and inflammatory responses, and a decrease in LPS-elicited homologous tolerance. We thus propose that NOD2 in the absence of MDP treatment might also play a negative regulatory role in the action of TLR4. Further, we demonstrated that both CARD and LRR domains of the NOD2 protein were responsible for the negative regulatory action on TLR4. In summary, it is the first time to demonstrate that NOD2 have dual effects on TLR4 signalling and exert a novel ligand-independent action. Elucidating molecular mechanisms by which NOD2 exerts its ligand-independent action on TLR4 requires further investigation.

Background The aim of this study was to reevaluate the clinical significance of sonographic appearances, in particular the application of color Doppler ultrasound imaging, in discriminating peripheral air-fluid lung abscess from empyema. Methods We retrospectively studied patients who had had peripheral air-fluid lesions due to empyema or lung abscess and who had undergone color Doppler ultrasound and grayscale ultrasound examinations between January 2003 and October 2007. A total of 34 patients with confirmed lung abscess and 30 patients with empyema were identified. The four sonographic characteristics observed and analyzed were the wall characteristics of the lesions (wall width, luminal margin, outer margin, and chest wall angle), split pleura sign, internal echogenicity (suspended microbubble sign, complex-septated effusions, and passive atelectasis), and identification of color Doppler ultrasound vessel signals in pericavitary lesions (consolidation or atelectasis). Results Among the sonographic characteristics, complex-septated effusions and passive atelectasis were specific for empyema, but the sensitivity was only 40% (n = 12 of 30) and 47% (n = 14 of 30), respectively. The identification of color Doppler ultrasound vessel signals in pericavitary consolidation was the most useful and specific for identifying lung abscesses. In our series, if we define the identification of color Doppler ultrasound vessel signals in a pericavitary consolidation as a predictor for peripheral lung abscess, we can achieve sensitivity, specificity, positive predictive value, and negative predictive value of 94%, 100%, 100%, and 94%, respectively. Conclusions Color Doppler ultrasound is a powerful tool for differentiating the peripheral air-fluid abscess from empyema, with high specificity and without any risk.

The aim of this study was to determine whether pleural fluid C-reactive protein (CRP) is useful in distinguishing complicated parapneumonic pleural effusion (CPPE) and empyema from uncomplicated parapneumonic pleural effusions (UPPE). A total of 69 consecutive patients with parapneumonic effusions were enrolled in the study: 29 with UPPE, 29 with CPPE, and 11 with empyema. Concentrations of standard biochemical parameters together with CRP in the pleural fluid were measured using an immunoturbidimetric assay. Pleural CRP was significantly higher in CPPE (11.6 mg/dl) and in empyema (12.2 mg/dl) than in UPPE (3.9 mg/dl). A cutoff value of 8.7 mg/dl for pleural CRP in the diagnosis of CPPE and empyema resulted in a sensitivity, specificity, and area under the receiver-operating characteristic curve (AUC) of 0.80, 0.97 and 0.94, respectively. Traditional lactic dehydrogenase (LDH)or = 1000 U/L and glucoseor = 60 mg/dl can differentiate CPPE and empyema from UPPE, with the sensitivity, specificity, and AUC achieving 0.75/0.60.1.00/1.00,0.95/0.22, respectively. However, for the detection of CPPE and empyema, the combination of pleural fluid CRPor = 8.7 mg/dl and LDHor = 1000 U/L was valuable in achieving a sensitivity, specificity, and AUC of 0.97/1,00/0.95. This study suggests that measurement of pleural CRP can be useful in the workup of patients with a parapneumonic effusion in order to differentiate CPPE from UPPE.

Rationale and Objectives Using low-dose computed tomography (LDCT), small and heterogeneous lung tumors are detected in screening. The criteria for assessing detected tumors are crucial for determining follow-up or resection strategies. The purpose of this study was to investigate the capacity of density features in differentiating lung tumors. Materials and Methods From July 2008 to December 2011, 48 surgically confirmed tumors (29 malignancies, comprising 17 cases of adenocarcinoma and 12 cases of adenocarcinoma in situ [AdIs], and 19 benignancies, comprising 11 cases of atypical adenomatous hyperplasia [AAH] and eight cases of benign non-AAH) in 38 patients were retrospectively evaluated, indicating that the positive predictive value (PPV) of physicians is 60.4% (29/48). Three types of density features, tumor disappearance rate (TDR), mean, and entropy, were obtained from the CT values of detected tumors. Results Entropy is capable of differentiating malignancy from benignancy but is limited in differentiating AdIs from benign non-AAH. The combination of entropy and TDR is effective for predicting malignancy with an accuracy of 87.5% (42/48) and a PPV of 89.7% (26/29), improving the PPV of physicians by 29.3%. The combination of entropy and mean adequately clarifies the four pathology groups with an accuracy of 72.9% (35/48). For tumors with a mean below −400 Hounsfield units, the criterion of an entropy larger than 5.4 might be appropriate for diagnosing malignancy. For others, the pathology is either benign non-AAH or adenocarcinoma; adenocarcinoma has a higher entropy than benign non-AAH, with the exception of tuberculoma. Conclusions Combining density features enables differentiating heterogeneous lung tumors in LDCT.

This paper proposes an efficient algorithm for detecting pleural objects that come in contact with a nodule. To reduce complexity, the algorithm recursively performed a curve-fitting method on each slice of the volume of interest to locate the object between the parietal and visceral pleurae surfaces and measured the quality of the fitting curve. When a nodule contacted the surfaces of the chest wall or diaphragm, they were automatically separated using the fitting curve. The algorithm was performed on 864 slices of 40 nodules. The segmentation results were visually inspected by a consensus of attending physicians to search for any segmentation errors. The consensus accepted 93.6% of the segmentation results.

To explore the diagnostic value of density features in lung tumor screened from low dose computed tomography (LDCT) with thin section. a computer-aided diagnosis (CAD) system was established to assist in defining tumor and density features. Forty-eight surgically confirmed tumors in 38 patients screened by thin-section LDCT were retrospectively enrolled in consecutive manner to examine the performance of this system. the confirmed surgical results included 29 malignant and 19 benign tumors. the pathology of malignancy were adenocarcinoma (AdCa, n=17) and adenocarcinoma in situ (AdIs, n=12). the benignancy included atypical adenomatous hyperplasia (AAH, n=11) and benign non-AAH (n=8). of density features, tumor Entropy provided the best power to differentiate malignancy from benignancy (p

Vascular endothelial growth factor C (VEGF-C) has been identified as a multifaceted factor participating in the regulation of tumor angiogenesis and lymphangiogenesis. VEGF-C is not only expressed in endothelial cells, but also in tumor cells. VEGF-C signaling is important for progression of various cancer types through both VEGF receptor-2 (VEGFR-2) and VEGF receptor-3 (VEGFR-3). Likewise, both receptors are expressed mainly on endothelial cells, but also expressed in tumor cells. The dimeric VEGF-C undergoes a series of proteolytic cleavage steps that increase the protein binding affinity to VEGFR-3; however, only complete processing, removing both the N- and C-terminal propeptides, yields mature VEGF-C that can bind to VEGFR-2. The processed VEGF-C can bind and activate VEGFR-3 homodimers and VEGFR-2/VEGFR-3 heterodimers to elicit biological responses. High levels of VEGF-C expression and VEGF-C/VEGFRs signaling correlate significantly with poorer prognosis in a variety of malignancies. Therefore, the development of new drugs that selectively target the VEGF-C/VEGFRs axis seems to be an effective means to potentiate anti-tumor therapies in the future.

Extracellular ATP is an important signaling molecule mediating quite divergent specific biological effects. Even though recent studies suggest a potential role of ATP in cancer progress, its real impact in chemotherapeutic efficacy remains unclear. In the present study, we investigated the effect of ATP on the cytotoxicity of doxorubicin in various cancer cell types and found that ATP had no effect on doxorubicin cytotoxicity in colon, prostate, breast, and cervical cancers or in osteosarcoma. In contrast, ATP has divergent effects on lung cancer cells: it can protect against doxorubicininduced cell death in non-metastatic lung cancer CL1.0 cells, but not in highly metastatic CL1.5 cells. Both apoptotic (characterized by sub-G1 peak, caspase 3 activation, poly(ADP-ribose) polymerase-1 cleavage) and necrotic (characterized by propidium iodide uptake and ROS production) features induced by doxorubicin in CL1.0 cells were reduced by ATP. In addition, ATP attenuated p53 accumulation, DNA damage (assessed by poly(ADP-ribose) formation and the comet assay) and topoisomerase II inhibition after doxorubicin treatment, and doxorubicin cytotoxicity was diminished by the p53 inhibitor pifithrin-a. Moreover, UTP, UDP, ADP, and pyrophosphate sodium pyrophosphate tetrabasic decahydrate diminished the antitumor effect of doxorubicin in CL1.0 cells, whereas purinergic P2 receptors antagonists did not abrogate the action of ATP. In summary, ATP fails to alter the antitumor efficacy of doxorubicin in most cancer cell types, except in CL1.0 cells, in which pyrophosphate mediates the cell protection afforded by ATP via attenuation of reactive oxygen species production, DNA damage, p53 accumulation, and caspase activation. (Cancer Sci 2012; 103: 1811‐1819)

Low dose CT (LDCT) is imperative in pursuing early detection of lung cancer. From October 2008 to September 2010, 25 patients with 31 nodules were found in the health examination and underwent surgical operations at China Medical University Hospital . Of which, the lesions is classified into atypical adenomatous hyperplasia (AAH), bronchioloalveolar carcinoma (BAC), malignant, or benign other than AAH in the pathologic results. In this study, objective and standardized features were defined based on the CT number analysis in order to describe lesions and construct a computer aided diagnosis system. The diagnostic performance of constructed system is 70.97% in the accuracy index.

A mass screening of lung function associated with air pollutants for children is limited. This study assessed the association between air pollutants exposure and the lung function of junior high school students in a mass screening program in Taipei city, Taiwan. Among 10,396 students with completed asthma screening questionnaires and anthropometric measures, 2919 students aged 12–16 received the spirometry test. Forced vital capacity (FVC) and forced expiratory flow in 1 s (FEV 1 ) in association with daily ambient concentrations of particulate matter with diameter of 10 μm or less (PM 10 ), sulfur dioxide (SO 2 ), carbon monoxide (CO), nitrogen dioxide (NO 2 ), and ozone (O 3 ) were assessed by regression models controlling for the age, gender, height, weight, student living districts, rainfall and temperature. FVC, had a significant negative association with short-term exposure to O 3 and PM 10 measured on the day of spirometry testing. FVC values also were reversely associated with means of SO 2 , O 3 , NO 2 , PM 10 and CO exposed 1 d earlier. An increase of 1-ppm CO was associated with the reduction in FVC for 69.8 mL (95% CI: −115, −24.4 mL) or in FEV 1 for 73.7 mL (95% CI: −118, −29.7 mL). An increase in SO 2 for 1 ppb was associated with the reductions in FVC and FEV 1 for 12.9 mL (95% CI: −20.7, −5.09 mL) and 11.7 mL (95% CI: −19.3, −4.16 mL), respectively. In conclusion, the short-term exposure to O 3 and PM 10 was associated with reducing FVC and FEV 1 . CO and SO 2 exposure had a strong 1-d lag effect on FVC and FEV 1 .

NOD2 of the NLRs and TLR4 of the TLRs are major pattern-recognition receptors, which sense different microbial pathogens and have important roles in innate immunity. Herein, we investigated the roles of NOD2 in TLR4-mediated signalling and gene regulation in RAW264.7 macrophages. We found that MDP (a NOD2 ligand) increased LPS-induced expressions of TNF-α, IL-1β, IL-6, iNOS and COX-2. MDP did not affect LPS-induced activation of MAPKs or IKK, while it potentiated LPS-induced NF-κB activation. Meanwhile TLR4 activation increased NOD2 mRNA expression, and upregulated NOD2 upon MDP treatment is a positive regulator of TLR4-mediated signalling. Intriguingly we found that NOD2 silencing led to increases in LPS-induced signal transduction and inflammatory responses, and a decrease in LPS-elicited homologous tolerance. We thus propose that NOD2 in the absence of MDP treatment might also play a negative regulatory role in the action of TLR4. Further, we demonstrated that both CARD and LRR domains of the NOD2 protein were responsible for the negative regulatory action on TLR4. In summary, it is the first time to demonstrate that NOD2 have dual effects on TLR4 signalling and exert a novel ligand-independent action. Elucidating molecular mechanisms by which NOD2 exerts its ligand-independent action on TLR4 requires further investigation.

Introduction: Given one third of the human population have been infected with tuberculosis, it is important to delineate the relationship between tuberculosis and lung cancer. This study explored whether contracting pulmonary tuberculosis is associated with an increased risk of developing lung cancers. Methods: In a cohort of 716,872 insured subjects, free from cancers, aged 20 years and older, 4480 patients with newly diagnosed tuberculosis were identified from the universal insurance claims in 1998–2000 and tracked until 2007 with the remaining insured without tuberculosis. We compared the incidence of lung cancers between the two cohorts and measured the associated hazard of developing lung cancer. Results: The incidence of lung cancers was approximately 11-fold higher in the cohort of patients with tuberculosis than nontuberculosis subjects (26.3 versus 2.41 per 10,000 person-years). Cox proportional hazard regression analysis showed a hazard ratio of 4.37 (95% confidence interval [CI]: 3.56–5.36) for the tuberculosis cohort after adjustment for the sociodemographic variables or 3.32 (95% CI: 2.70–4.09) after further adjustment for chronic obstructive pulmonary disease (COPD), smoking-related cancers (other than lung cancer), etc. The hazard ratio increased to 6.22 (95% CI: 4.87–7.94) with the combined effect with COPD or to 15.5 (95% CI: 2.17–110) with the combined effect with other smoking-related cancers. Conclusions: This study provides a compelling evidence of increased lung cancer risk among individuals with tuberculosis. The risk may increase further with coexisting COPD or other smoking-related cancers.

The aim of this study was to reevaluate the clinical significance of sonographic appearances, in particular the application of color Doppler ultrasound imaging, in discriminating peripheral air-fluid lung abscess from empyema.We retrospectively studied patients who had had peripheral air-fluid lesions due to empyema or lung abscess and who had undergone color Doppler ultrasound and grayscale ultrasound examinations between January 2003 and October 2007. A total of 34 patients with confirmed lung abscess and 30 patients with empyema were identified. The four sonographic characteristics observed and analyzed were the wall characteristics of the lesions (wall width, luminal margin, outer margin, and chest wall angle), split pleura sign, internal echogenicity (suspended microbubble sign, complex-septated effusions, and passive atelectasis), and identification of color Doppler ultrasound vessel signals in pericavitary lesions (consolidation or atelectasis).Among the sonographic characteristics, complex-septated effusions and passive atelectasis were specific for empyema, but the sensitivity was only 40% (n = 12 of 30) and 47% (n = 14 of 30), respectively. The identification of color Doppler ultrasound vessel signals in pericavitary consolidation was the most useful and specific for identifying lung abscesses. In our series, if we define the identification of color Doppler ultrasound vessel signals in a pericavitary consolidation as a predictor for peripheral lung abscess, we can achieve sensitivity, specificity, positive predictive value, and negative predictive value of 94%, 100%, 100%, and 94%, respectively.Color Doppler ultrasound is a powerful tool for differentiating the peripheral air-fluid abscess from empyema, with high specificity and without any risk.

SummaryAim: The retention index, a traditionally quantitative analysis of two-phase 201Tl single photon emission computed tomography (SPECT) of the chest, is manually calculated by experienced physicians from comparable 2-D ROI. However, a 3-D ROI would provide more information than a 2-D ROI extracted from a single frame of SPECT. We propose a new diagnostic system, computer-aided diagnosis (CAD), to automatically detect suspicious lesions as 3-D objects on chest 201Tl-SPECT, and assist the physician in interpreting these images. Patients, methods: Seventy patients with thoracic lesions and confirmed diagnoses were enrolled to test this automatic CAD system. The reliability of the CAD system in detecting lesions as 3-D objects was compared to the 2-D ROI of 201Tl-SPECT found by the manually visualized method. Furthermore, we also proposed a novel index, the retention index using the heart (RIH), to differentiate high retention (slow clearance, increasing target to heart ratio) as a criterion for a malignant lesion, from low retention (faster clearance, small or no increase of the target to heart ratio) for benign lesions. Results: The CAD system can achieve a detection rate of 100% in automatically searching for thoracic lesions in 201Tl-SPECT. In diagnostic performance, the CAD system with the RIH of comparable 3-D objects has an area under the ROC curve of 0.86, higher than the 0.78 of the traditional RI method (p = 0.198). Conclusion: The CAD system of two-phase 201Tl-SPECT is a promising tool for detecting and diagnosing thoracic lesions with a diagnostic accuracy of 0.81.

Dynamic flow ultrasound (DFUS) is a new color Doppler imaging method with better B-mode imaging and fewer blooming effects and color noises. This study was designed to compare the imaging quality of vessel signals in thoracic lesions using DFUS, color Doppler US (CDUS), and power Doppler US (PDUS).Thirty-four patients with thoracic lesions abutting pulsatile organs [heart (n = 13), aorta (n = 14) and pulmonary artery (n = 7)] and undergoing complete chest US examinations were included to assess the imaging quality about vessel signals, blooming effect, color noise, and the influence of decision in needle biopsy between different US modes.Our results showed that DFUS, CDUS, and PDUS could all demonstrate the vessel signals clearly (all P.05). However, when focusing on the blooming effect and color noise, DFUS showed the more superior imaging quality than CDUS and PDUS (all Por = .001); and acceptable blooming effects/color noise were found with 100% (34/34)/97% (33/34), 35% (12/34)/68% (23/34), and 26% (9/34)/38% (13/34) in DFUS, CDUS, and PDUS, respectively. Especially, in the assessment of decision making for percutaneous needle biopsy, DFUS had the less influence than CDUS and PDUS (3% [1/33] versus 29% [10/34] and 3% [1/33] versus 38% [13/34], both P.01).We concluded that DFUS has a clearly more superior imaging quality than CDUS and PDUS in demonstrating the vessel signals of thoracic lesions, with less blooming effect and color noise.

Objective. The purpose of this study was to evaluate the role of the sonographic features of lymphocyte-rich exudative pleural effusions in the differential diagnosis of tuberculosis and lung cancer in an area with a high incidence of tuberculosis. Methods. Medical records of patients undergoing chest sonography between January 2003 and June 2005 (30 months) were reviewed retrospectively. The enrolled patients included 73 with lung cancer-related pleural effusions and 93 with tuberculous pleural effusions. The sonographic appearances of the pleural effusions were defined in terms of 4 patterns: anechoic, homogeneously echogenic, complex septated, and complex nonseptated. Results. Among the 73 lung cancer-related pleural effusions, there were sonographic appearances of an anechoic pattern in 11% (8/73), a complex septated pattern in 4% (3/73), and a complex nonseptated pattern in 85% (62/73). In 93 tuberculous pleural effusions, there were sonographic appearances of an anechoic pattern in 12% (11/93), a complex septated pattern in 47% (44/93), and a complex nonseptated pattern in 41% (38/93). Apparently, a complex septated pattern in the sonographic appearance of lymphocyte-rich pleural effusions is a useful diagnostic predictor for differentiating tuberculosis from lung cancer (95% confidence interval, ‐0.57 to ‐0.29). If we define the complex septated pattern in the sonographic appearance of lymphocyte-rich exudative pleural effusions as a predictor for tuberculous pleural effusions, we can achieve sensitivity, specificity, positive predictive value, negative predictive value, and positive likelihood ratio values of 47%, 96%, 94%, 59%, and 12, respectively. Conclusions. A complex septated pattern in the sonographic appearance is a useful predictor of tuberculosis in lymphocyte-rich exudative pleural effusions. Key words: lung cancer; lymphocyte-rich exudative effusions; septation; sonography; tuberculosis.

The lung cancer blood supply originates from the bronchial artery. If vessel signals within pulmonary lesions can be confirmed to be those of the pulmonary artery, color Doppler ultrasound (US) should be able to predict and differentiate benign lesions from lung cancers. Two hundred sixty-four patients with abutting thoracic lesions (including 125 lung cancers and 139 benign lesions) underwent color Doppler US examinations. A pulsatile flow, with the vessel signal length on sonographic appearance ≥1 cm demonstrated by color Doppler US, was arbitrarily defined as a pulmonary artery vessel signal. Of the 264 thoracic lesions, 73 (58%) lung cancers and 107 (77%) benign lesions had detectable color Doppler US pulsatile flow vessel signals. Analyzing the pulsatile flow vessel signals, the color Doppler US pulmonary artery vessel signal was present in 74 (53%) benign lesions, but was found in only two (2%) lung cancers of a specific alveolar cell carcinoma with lobar consolidation. Using the pulmonary artery vessel signal, color Doppler US can be valuable in predicting and differentiating benign lesions from lung cancers (p < 0.0001, sensitivity = 0.53, specificity = 0.98 and positive likelihood ratio 26.5). In conclusion, color Doppler US pulmonary artery vessel signal sign is useful in predicting and differentiating benign lesions from lung cancers. (E-mail: hsuwh@www.cmuh.org.tw)

Background: This study examines the usefulness of thallium-201 single photon emission computed tomography ( 2 0 1 TI SPECT) in differentiating pulmonary malignancies from benign lesions by using dual phase 2 0 1 TI scintigraphy. Methods: One hundred and six patients with thoracic lesions and confirmed diagnoses were assessed by 2 0 1 TI chest SPECT examinations; of these 106 enrolled thoracic lesions, 59 were malignancies and 47 were benign lesions. Dual phase 2 0 1 TI SPECT was performed with an early image acquired at 10-20 min and a delayed image at 2-3 h after intravenous injection of 2-3 mCi of 2 0 1 TI. The results of 2 0 1 TI SPECT images were classified as either malignant or benign lesions by visual interpretation: the lesion was interpreted as positive for malignancy if the uptake of 2 0 1 TI in the lung lesion in the delayed phase was increased or persistent as compared with that in the early phase image; otherwise, it was considered as a benign lesion. Simultaneously, the traditional method of retention index (RI) was also calculated to help in the differential diagnosis of pulmonary lesions. Then, both methods of dual phase 2 0 1 TI SPECT, visual reading and traditional RI, were compared to differentiate pulmonary malignancies from benign lesions. Results: Analyzing the image results, we found that dual phase 2 0 1 TI SPECT could differentiate pulmonary malignancies from benign lesions with a sensitivity of 83%, a specificity of 91% and an accuracy of 87%. Moreover, by using the traditional RI method of 2 0 1 TI SPECT, it could differentiate pulmonary malignancy from benign lesions with a sensitivity of 79.3%, a specificity of 80.8% and an accuracy of 80%. Conclusions: We conclude that using dual phase 2 0 1 TI SPECT with visual interpretation is a simpler and potentially more effective method for differentiating pulmonary malignancies from benign lesions, with results compatible with the traditional RI method of 2 0 1 TI SPECT.