Injectable hydrogel with doxorubicin-loaded ZIF-8 nanoparticles for tumor postoperative treatments and wound repair.

The need for tumor postoperative treatments aimed at recurrence prevention and tissue regeneration have raised wide considerations in the context of the design and functionalization of implants. Herein, an injectable hydrogel system encapsulated with anti-tumor, anti-oxidant dual functional nanoparticles has been developed in order to prevent tumor relapse after surgery and promote wound repair. The utilization of biocompatible gelatin methacryloyl (GelMA) was geared towards localized therapeutic intervention. Zeolitic imidazolate framework-8@ceric oxide (ZIF-8@CeO2, ZC) nanoparticles (NPs) were purposefully devised for their proficiency as reactive oxygen species (ROS) scavengers. Furthermore, injectable GelMA hydrogels loaded with ZC NPs carrying doxorubicin (ZC-DOX@GEL) were tailored as multifunctional postoperative implants, ensuring the efficacious eradication of residual tumor cells and alleviation of oxidative stress. In vitro and in vivo experiments were conducted to substantiate the efficacy in cancer cell elimination and the prevention of tumor recurrence through the synergistic chemotherapy approach employed with ZC-DOX@GEL. The acceleration of tissue regeneration and in vitro ROS scavenging attributes of ZC@GEL were corroborated using rat models of wound healing. The results underscore the potential of the multifaceted hydrogels presented herein for their promising application in tumor postoperative treatments.

Induction immune-checkpoint inhibitors for resectable oncogene-mutant NSCLC: A multicenter pooled analysis.

Despite limited efficacy of immunotherapy for advanced non-small-cell lung cancer (NSCLC) with driver mutations, whether neoadjuvant immunotherapy could be clinically valuable in those patients warrants further investigation. We utilized 40 oncogene-mutant NSCLC treated with induction immunotherapy from a large consecutive multicenter cohort. Overall response rate was 62.5% while 2 patients had disease progression. Of 39 patients that received surgery, R0 resection rate was 97.4%. The major pathological response (MPR) rate was 37.5% and the pathological complete response (pCR) rate was 12.5%. Pre-treatment PD-L1 expression was not a predictive biomarker in these patients. Median disease-free survival for all oncogenic mutation and EGFR mutation was 28.5 months. Indirect comparison through integrating CTONG1103 cohort showed neoadjuvant immunotherapy plus chemotherapy yielded the most superior efficacy among erlotinib and chemotherapy for resectable EGFR-mutant NSCLC. No MPR patients were identified with neoadjuvant immunotherapy plus chemotherapy for uncommon EGFR insertion or point mutations. Our results indicated the potential clinical feasibility of neoadjuvant immunotherapy for resectable localized oncogene-mutant NSCLC especially for EGFR-mutant NSCLC.

METTL3-induced DLGAP1-AS2 promotes non-small cell lung cancer tumorigenesis through m6A/c-Myc-dependent aerobic glycolysis.

The critical roles of N6-methyladenosine (m6A) modification have been demonstrated by more and more evidence. However, the cross talk of m6A and long noncoding RNAs (lncRNAs) in non-small cell lung cancer (NSCLC) tumorigenesis is still unclear. Here, this work focused on the functions and molecular mechanism of m6A-modified lncRNA DLGAP1 antisense RNA 2 (DLGAP1-AS2) in NSCLC. Microarray analysis found that lncRNA DLGAP1-AS2 is upregulated in NSCLC cells. Clinical data showed that DLGAP1-AS2 high-expression was correlated with advanced pathological stage and poor prognosis. Functionally, DLGAP1-AS2 overexpression promoted the aerobic glycolysis and DLGAP1-AS2 knockdown suppressed the tumor growth of NSCLC cells. Mechanistically, m6A methyltransferase METTL3 enhanced the stability of DLGAP1-AS2 via m6A site binding. Moreover, DLGAP1-AS2 interacted with YTHDF1 to enhance the stability of c-Myc mRNA through DLGAP1-AS2/YTHDF1/m6A/c-Myc mRNA. In conclusion, our work indicates the functions of m6A-modified DLGAP1-AS2 in the NSCLC aerobic glycolysis, disclosing a potential m6A-dependent manner for NSCLC treatment.

Plasma CXCL14 as a Candidate Biomarker for the Diagnosis of Lung Cancer.

Background: Effective biomarkers for early diagnosis of lung cancer are needed. Previous studies have indicated positive associations between abnormal circulating cytokines and the etiology of lung cancer. Methods: Blood samples were obtained from 286 patients with pretreatment lung cancer and 80 healthy volunteers. Circulating cytokine levels were detected with a Luminex assay and enzyme-linked immunosorbent assay (ELISA). Urine samples were obtained from 284 patients and 122 healthy volunteers. CXC chemokine ligand 14 (CXCL14) expression in tumors and nontumor regions of lung tissues from 133 lung cancer cases was detected by immunohistochemical (IHC) staining and immunofluorescence (IF) staining of formalin fixed paraffin-embedded (FFPE) tissues. Results: Compared with healthy volunteers, a 65.7-fold increase was observed in the level of CXCL14 in the plasma of lung cancer patients, and a 1.7-fold increase was observed in the level of CXCL14 in the urine of lung cancer patients, achieving a 0.9464 AUC (area under the curve) value and a 0.6476 AUC value for differentiating between lung cancer patients and healthy volunteers, respectively. Stromal CXCL14 expression was significantly associated with advanced pathologic stage (P<0.001), pathologic N stage (P<0.001), and recurrence and metastasis (P=0.014). Moreover, multivariate analysis suggested stromal CXCL14 expression as an independent predictor of DFS and OS. Conclusions: Our study demonstrates that CXCL14 might serve as a potential diagnostic and prognostic biomarker in patients with lung cancer. Impact: CXCL14 might serve as a potential diagnostic and prognostic biomarker in patients with lung cancer.

Urinary malate dehydrogenase 2 is a new biomarker for early detection of non-small-cell lung cancer.

Reliable and noninvasive biomarkers for the early diagnosis of non-small-cell lung cancer (NSCLC) are an unmet need. This study aimed to screen and validate potential urinary biomarkers for the early diagnosis of NSCLC. Using protein mass spectrometry, urinary MDH2 was found to be abundant both in patients with lung cancer and lung cancer model mice compared with controls. Urine samples obtained as retrospective and prospective cohorts including 1091 NSCLC patients and 736 healthy controls were measured using ELISA. Patients with stage I NSCLC had higher urinary MDH2 compared with healthy controls. The area under the receiver-operating characteristic curve (AUC) for the urinary MDH2 was 0.7679 and 0.7234 in retrospective and prospective cohorts to distinguish stage I cases from controls. Urinary MDH2 levels correlated with gender and smoking history. MDH2 expression levels were elevated in lung cancer tissues. MDH2 knockdown using shRNA inhibited the proliferation of lung cancer cells. Our study demonstrated that urinary MDH2 concentration was higher in early-stage NSCLC patients compared with that in controls and that MDH2 could serve as a potential biomarker for early detection of NSCLC.

Detection of circulating genetically abnormal cells in peripheral blood for early diagnosis of non-small cell lung cancer.

BACKGROUND: Circulating genetically abnormal cells (CACs) with specific chromosome variations have been confirmed to be present in non-small cell lung cancer (NSCLC). However, the diagnostic performance of CAC detection remains unclear. This study aimed to evaluate the potential clinical application of the CAC test for the early diagnosis of NSCLC. METHODS: In this prospective study, a total of 339 participants (261 lung cancer patients and 78 healthy volunteers) were enrolled. An antigen-independent fluorescence in situ hybridization was used to enumerate the number of CACs in peripheral blood. RESULTS: Patients with early-stage NSCLC were found to have a significantly higher number of CACs than those of healthy participants (1.34 vs. 0.19; P < 0.001). The CAC test displayed an area under the receiver operating characteristic (ROC) curve of 0.76139 for discriminating stage I NSCLC from healthy participants with 67.2% sensitivity and 80.8% specificity, respectively. Compared with serum tumor markers, the sensitivity of CAC assays for distinguishing early-stage NSCLC was higher (67.2% vs. 48.7%, P < 0.001), especially in NSCLC patients with small nodules (65.4% vs. 36.5%, P = 0.003) and ground-glass nodules (pure GGNs: 66.7% vs. 40.9%, P = 0.003; mixed GGNs: 73.0% vs. 43.2%, P < 0.001). CONCLUSIONS: CAC detection in early stage NSCLC was feasible. Our study showed that CACs could be used as a promising noninvasive biomarker for the early diagnosis of NSCLC. KEY POINTS: What this study adds: This study aimed to evaluate the potential clinical application of the CAC test for the early diagnosis of NSCLC. Significant findings of the study: CAC detection in early stage NSCLC was feasible. Our study showed that CACs could be used as a promising noninvasive biomarker for the early diagnosis of NSCLC.

Risk Prediction for Locoregional Recurrence in Epidermal Growth Factor Receptor-Mutant Stage III-pN2 Lung Adenocarcinoma after Complete Resection: A Multi-center Retrospective Study.

Background: This study aimed to develop a predictive model based on the risk of locoregional recurrence (LRR) in epidermal growth factor receptor (EGFR)-mutant stage III-pN2 lung adenocarcinoma after complete resection. Methods: A total of 11,020 patients with lung surgery were screened to determine completely resected EGFR-mutant stage III-pN2 lung adenocarcinoma. Patients were excluded if they received preoperative therapy or postoperative radiation therapy (PORT). The time from surgery to LRR was recorded. Clinicopathological variables with statistical significance predicting LRR in the multivariate Cox regression were incorporated into the competing risk nomogram. Patients were then sub-grouped based on different recurrence risk as a result of the nomogram. Results: Two hundred and eighty-eight patients were enrolled, including 191 (66.3%) with unforeseen N2 (IIIA1-2), 75 (26.0%) with minimal/single station N2 (IIIA3), and 22 (7.6%) with bulky and/or multilevel N2 (IIIA4). The 2-year overall cumulative incidence of LRR was 27.2% (confidence interval [CI], 16.3%-38.0%). IIIA4 disease (hazard ratio, 2.65; CI, 1.15-6.07; P=0.022) and extranodal extension (hazard ratio, 3.33; CI, 1.76-6.30; P<0.001) were independent risk factors for LRR and were incorporated into the nomogram. Based on the nomogram, patients who did not have any risk factor (low-risk) had a significantly lower predicted 2-year incidence of LRR than those with any of the risk factors (high-risk; 4.6% vs 21.9%, P<0.001). Conclusions: Pre-treatment bulky/multilevel N2 and pathological extranodal extension are risk factors for locoregional recurrence in EGFR-mutant stage III-pN2 lung adenocarcinoma. Intensive adjuvant therapies and active follow-up should be considered in patients with any of the risk factors.

LncRNA H19 downregulation confers erlotinib resistance through upregulation of PKM2 and phosphorylation of AKT in EGFR-mutant lung cancers.

First-generation EGFR tyrosine kinase inhibitors (TKIs) such as erlotinib have significant activity in NSCLC patients with activating EGFR mutations. However, EGFR-TKI resistance inevitably occurs after approximately 12 months of treatment. Acquired mechanisms of resistance, other than secondary mutations in EGFR (T790 M) which account for 50-60%, are less well understood. Here, we identified lncRNA H19 as a significantly downregulated lncRNA in vitro models and clinical specimens with acquired EGFR-TKI resistance, H19 knockdown or overexpression conferred resistance or sensitivity, respectively, both in vitro and in vivo models. H19 downregulation contributed to erlotinib resistance through interaction and upregulation of PKM2, which enhanced the phosphorylation of AKT. AKT inhibitors restored the sensitivity of erlotinib-resistant cells to erlotinib. In EGFR-mutant patients treated with EGFR-TKIs, low H19 levels were associated with a shorter progression-free survival (PFS) (P = 0.021). These findings revealed a novel mechanism of low-level H19 in the regulation of erlotinib resistance in EGFR-mutant lung cancers. Combination of AKT inhibitors and EGFR-TKIs could be a rational therapeutic approach for some subgroups of EGFR-mutant lung cancer patients.

Prognostic value of combined platelet, fibrinogen, neutrophil to lymphocyte ratio and platelet to lymphocyte ratio in patients with lung adenosquamous cancer.

The aim of the present study was to investigate the prognostic value of the combined platelet (PLT), fibrinogen (FBG), neutrophil to lymphocyte ratio (NLR) and platelet to lymphocyte ratio (PLR) (CO-NPF) for postoperative outcomes in patients with lung adenosquamous cancer (ASC). Test results from patients who presented at The Cancer Institute and Hospital of Tianjin Medical University between January 2005 and December 2013 were retrospectively reviewed. CO-NPF was scored between 0 and 4 according to increased PLT, FBG, NLR and PLR prior to being split into two groups based on the presence (≥2) or absence (<2) of the combination of increased inflammatory indexes. In total, data from 134 patients with ASC were reviewed for the present study. Multivariate analysis identified that increased CO-NPF (P=0.001 and P<0.001, respectively), PLR (P=0.011 and P=0.001, respectively) and FBG (P=0.001 and P<0.001, respectively) were independently associated with shorter disease-free survival (DFS) and overall survival (OS). NLR (P=0.006) and PLT (P=0.001) were independent prognostic factors for OS. The area under the receiver operating characteristic curves of CO-NPF (area under the curve, 0.652, P=0.008, 95% confidence interval, 0.551-0.752) was increased compared with NLR, PLR, PLT and FBG individually, suggesting that CO-NPF has greater predictive value. CO-NPF was significantly and independently associated with shorter DFS and OS, and had greater predictive value compared with NLR, PLR, PLT and FBG in patients with ASC who underwent surgery.

[Prognostic analysis of 450 patients with lung squamous cancer].

OBJECTIVE: To study the treatment strategy and prognosis and its affected factors of lung squamous cancer according retrospective analysis. METHODS: Clinic data of 450 lung squamous cancer inpatient cases who were performed complete resection from January 2004 to January 2007, was retrospectively reviewed. There were 363 male and 87 female patients, aged from 31 to 82 years, with a mean of 60.5 years and a median of 62 years. RESULTS: The overall 5-year survival rate was 52.4%. Cox Regression suggested that preoperative N status (χ(2) = 18.969, P = 0.000), N stage (χ(2) = 44.069, P = 0.000) and TNM stage (χ(2) = 63.025, P = 0.000) are independent factors affecting the prognosis. Adjuvant chemotherapy affects the prognosis of stage II-IIIA lung squamous cancer (5-year survival rate: 48.9% vs. 37.7%, χ(2) = 3.946, P = 0.047). Studying the combined therapy of stage IIIA, the chemoradiotherapy group achieved the best survival (48.8%), then single chemotherapy group (35.9%) and no treatment group (28.5%), and the single radiotherapy group achieved the poorest survival rate (11.1%), and there were statistically significant differences among them (χ(2) = 8.397, P = 0.038). CONCLUSIONS: The 5-year survival rate of lung squamous cancer has significantly increased through promoting the standard of operation, especially increasing the standard of lymph node dissection. Adjuvant chemotherapy is benefit for stage II-IIIA patients and combined chemoradiotherapy is the best choice for stage IIIA patients. If preoperative examination suggests mediastinal lymph node's enlargement and fusion, the operation should not be performed.

Value of the metastatic lymph node ratio for predicting the prognosis of non-small-cell lung cancer patients.

BACKGROUND: The aim of this study was to investigate the relation between the metastatic lymph node ratio (LNR) and the prognosis of non-small-cell lung cancer (NSCLC). METHODS: A total of 301 patients with N1 or N2 NSCLC who underwent complete pulmonary resection were analyzed retrospectively. The correlations between the LNR and clinical and pathologic data were analyzed using χ(2) test analysis. The prognostic value of the LNR was calculated by univariate Kaplan-Meier survival analysis and multivariate Cox proportional hazard model analysis. The risk groups were classified by a combination of the LNR and pN stage. RESULTS: The LNR was correlated with age, smoking status, pathologic type, subcarinal lymph node, clinical staging, N stage (P < 0.05), and the number of positive lymph nodes and positive lymph node stations (P < 0.0001). In the univariate analysis, the LNR played an important role in predicting overall survival (OS) (P < 0.0001) and disease-free survival (P < 0.0001) by Kaplan-Meier survival analysis. In the multivariate analysis, high LNR (>18%) was an independent poor prognostic factor for OS [hazard ratio (HR) 2.5034, 95% confidence interval (CI) 1.6096-3.8933, P < 0.0001] and DFS (HR 1.9023, 95% CI 1.2465-2.9031, P = 0.0031). Stratification into high-, medium-, and low-risk groups-based on high-risk factors (LNR > 18%, N2) intermediate-risk factors (LNR > 18%, N1 or LNR < 18%, N2), and low-risk factors (LNR < 18%, N1)-could efficiently predicted outcomes (P < 0.0001) of patients with lymph node-positive NSCLC. CONCLUSIONS: The combination of the LNR and pN status provides a valuable help with prognosis. However, these results must be evaluated further in a large prospective randomized clinical trial.

[Treatment and prognostic analysis of 1638 patients with non-small cell lung cancer].

OBJECTIVE: To study the prognosis and prognostic factors of non-small-cell lung carcinoma (NSCLC) according the new TNM stage system. METHODS: Clinic data of 1638 inpatient cases admitted from January 2001 to January 2005 were retrospectively reviewed. There were 1083 male and 555 female patients in the study and the average age was 59.5 years. All the patients received surgical procedures. RESULTS: The overall 1, 3, 5-year survival rate was 80.0%, 52.3%, 39.0%. The main prognostic factors were bronchial stump, operation type, T stage, N stage, the number of lymph nodes (LNs) in lymph nodes dissection (1 - 10, 11 - 20, and > 20), overall N stations (< 4 and ≥ 4) and postoperative radiotherapy (all P < 0.05). Cox regression suggested that T stage (P = 0.000), N stage (P = 0.000), operation type (P = 0.001) and LNs (P = 0.013) were independent factors affecting the prognosis. CONCLUSIONS: The overall survival rate of NSCLC is poor. T stage, N stage, operation type and LNs are independent factors affecting the prognosis.

Up-regulation of the expression of S100A8 and S100A9 in lung adenocarcinoma and its correlation with inflammation and other clinical features.

BACKGROUND: S100A8 and S100A9 are two members of the S100 protein family characterized by the presence of two Ca2+-binding sites of the EF-hand type. Previous studies suggested that the whole S100 family displays significant functions in tumor growth, progression and invasion. This study aimed to determine the expression of the two indices of the family, S100A8 and S100A9, in lung cancer tissues and normal lung tissues and its correlation with clinical features. METHODS: A total of 60 cases with a variety of clinical data that were diagnosed with different histological subtypes of lung cancer were investigated. Semi-quantitative reverse transcriptase-PCR (Sq-Rt-PCR) and immunohistochemical staining of cancer, adjacent and peripheral lung tissues were executed to distinguish the expression patterns of S100A8 and S100A9 and to further clarify their correlation with clinical features. RESULTS: Immunohistochemical staining of both proteins showed a significant up-regulation in lung cancer tissue (S100A8, S100A9, P<0.0001), and PCR revealed that the levels of S100A8 and S100A9 expression were significantly higher in lung cancer tissues (S100A8 P=0.002/0.004; S100A9 P=0.022/0.026). The higher expression was found to be correlated with the clinical characteristics of adenocarcinoma, inflammation and stage IV lesion. CONCLUSIONS: S100A8, S100A9 up-regulation was found in the lung adenocarcinoma and end stage lung cancer tissue, the correlation of which with their higher expression in inflammatory lung tissues may indicate the collaborative effect of inflammation on the progression of cancer.

[A clinical and prognostic retrospective analysis of IIIA-N2 non-small cell lung cancer].

OBJECTIVES: To analyze the clinical conditions of postoperative patients with IIIA-N2 non-small cell lung cancer (NSCLC) and the prognostic factors related with survival of NSCLC, and to investigate the influence of operation and therapy on prognosis. METHODS: Clinical data of 657 inpatient cases with IIIA-N2 NSCLC admitted from January 2000 to December 2005 was retrospectively reviewed. The Kaplan-Meier method was used for survival analysis. The Log-rank law was applied to analyze the relationship between the variables and the prognosis in monovariate analysis, while Cox proportional hazard regression model was used to make multivariate analysis. RESULTS: The 1-, 3-and 5-year accumulative survival rates of the operative patience were 64.4%, 26.0% and 17.9%, respectively. The median survival time was 18 months. In monovariate analysis, the main unfavorable factors that affect life span involve were the diameter of tumor, T stage, skip metastasis of N2 lymph node, the number of metastatic lymph nodes, the metastasis of subcarinal lymph nodes, adjuvant chemotherapy, the cycle of adjuvant chemotherapy, postoperative radiotherapy, and the modality of therapy (the effect of naive surgery was disappointed, while the prognosis of the patients with adjuvant chemoradiotherapy was better than those with chemotherapy alone). A multivariate analysis using Cox regression identified 5 factors of prognosis: the diameter of tumor (P = 0.001), the metastasis of subcarinal lymph nodes (P = 0.019), the number of metastatic lymph nodes (P = 0.006), the cycle of adjuvant chemotherapy (P = 0.007), postoperative radiotherapy (P = 0.055), and adjuvant chemoradiotherapy (P = 0.026). CONCLUSIONS: The 5-year survival rate of the patients with IIIA-N2 Non-small cell lung cancer is poor. Tumor size, the metastasis of subcarinal lymph nodes, the number of metastatic LNs, the cycle of adjuvant chemotherapy, and postoperative radiotherapy have an effect on the prognosis. The prognosis of postoperative patients with single-level N2 and multi-level N2 disease is similar, and the key point of survival is the number of nodes involved. The therapeutic effect of patience given adjuvant chemoradiotherapy is superior to those treated with adjuvant chemotherapy.

[Value of thyroid transcription factor-1 in identification of the prognosis of bronchioloalveolar carcinoma].

OBJECTIVE: To study the expression and clinical significance of cytokeratin subtypes CK7 and CK20 and thyroid transcription factor-1 (TTF-1) in bronchioloalveolar carcinoma (BAC), and to investigate the value of these factors in identification of the prognosis of BAC. METHODS: Eighty-one specimens of BAC resected during operation, 68 of non-mucinous type and 13 of mucinous type, underwent immunohistochemical examination to detect the expression of CK7, CK20, and TTF-1. The value of these 3 factors in the identification of the prognosis of BAC was examined by survival analysis. RESULTS: All specimens showed positive expression of CK7, CK20, and TTF-1. There was no significant differences in CK7 expression rate among different ages, clinical stages, and pathological subtypes (all P > 0.05). Compared with that in the BAC of stage III and that of BAC of non-mucinous type, the CK20 positive rates of the BAC of stage I - II and mucinous type were both significantly higher (chi(2) = 3.928, P < 0.05, and chi(2) = 11.512, P < 0.05). The TTF-1 positive rates of the BAC of stage III and nonmucinous type were significantly higher than those of stage I - II and mucinous type respectively (chi(2) = 7.840, P < 0.05, and chi(2) = 19.497, P < 0.05). There was a statistically significant positive correlation between the expression of CK7 and expression of TTF-1 (r = 0.257, P = 0.021). Univariate analysis showed that the main prognostic factors were TTF-1 expression (P = 0.017), clinical stage (P = 0.000), tumor diameter (P = 0.017) and N stage (P = 0.000). Strata analysis suggested that in the nonmucinous type BAC patients the survival time of those positive for TTF-1 expression was superior to those negative for TTF-1 (P = 0.009); and in the stage III BAC patients the survival time of those positive for TTF-1 was superior to those negative for TTF-1 (P = 0.022). Cox regression analysis suggested that TTF-1 (P = 0.035), TNM stage (P = 0.000), tumor diameter (P = 0.034), and N stage (P = 0.000) were independent factors affecting the prognosis. CONCLUSION: There is a statistically significant positive correlation between the expression of CK7 and the expression of TTF-1. TTF-1 and CK20 provide a new evidence for the molecular staging of BAC. TTF-1, TNM stage, tumor diameter and N stage are all independent factors affecting the prognosis of BAC.