Expert Consensus on Prevention and Treatment of COVID-19 Infection in Patients with Lung Cancer

Corona virus disease 2019(COVID-19)infection has become a major public health issue affecting human health.The main goal of epidemic prevention and control at the current stage in China is to“protect people’s health and prevent severe cases”.Patients with lung cancer who receive antitumor therapy have low immunity,and the risk of severe illness and death once infected is much higher than healthy people,so they are vulnerable to COVID-19 infection.At present,less attention has been paid to the prevention and treatment of COVID-19 infection in patients with lung cancer in domestic guidelines and consensus.Based on the published data in China and abroad,we proposed recommendations and formed expert consensus on the vaccination of COVID-19,the use of neutralizing antibodies and small molecule antiviral drugs for patients with lung cancer,for physician’s reference.

Bioinformatics Identification of ZNFs/LINC00520/miR-181d/BCL2 Axis as a Novel Network in Cisplatin-Resistant Lung Adenocarcinoma Cells

Background: Resistance to cisplatin (DDP) leads to poor prognosis in patients with Lung Adenocarcinoma (LUAD) and limits its clinical application. It has been confirmed that autophagy promotes chemoresistance and, therefore, novel strategies to reverse chemoresistance by regulating autophagy are desperately needed. Methods: The differentially expressed lncRNAs (DElncRNAs), miRNAs (DEmiRNAs), and mRNAs (DEmRNAs) between A549 and A549/DDP cell lines were identified using the limma package in R, after gene expression profiles were obtained from Gene Expression Omnibus (GEO) database. By combining Autophagy-Related Genes (ARGs) from Human Autophagy Database (HADb), the interactions lncRNA-miRNAs and the interactions miRNAs-mRNAs respectively predicted by miRcode and miRDB/Targetscan database, the autophagy-related ceRNA network was constructed. Then, extraction of ceRNA subnetwork and Cox regression analyses were performed. A prognosis-related ceRNA subnetwork was constructed, and the upstream Transcription Factors (TFs) regulating lncRNAs were predicted by the JASPAR database. Finally, the expression patterns of candidate genes were further verified by quantitative real-time polymerase chain reaction (qRT-PCR) experiments. Results: A total of 3179 DEmRNAs, 180 DEmiRNAs, and 160 DElncRNAs were identified, and 35 DEmRNAs were contained in the HADb. Based on the ceRNA hypothesis, we established a ceRNA network, including 10 autophagy-related DEmRNAs, 9 DEmiRNAs, and 14 DElncRNAs. Then, LINC00520, miR-181d, and BCL2 were identified to construct a risk score model, which was confirmed to be a well-predicting prognostic factor. Furthermore, 5 TF ZNF family members were predicted to regulate LINC00520, whereas the RT-PCR results showed that the 5 ZNFs were consistent with the bioinformatics analysis. Finally, a ZNF regulatory LINC00520/miR-181d/BCL2 ceRNA subnetwork was constructed. Conclusions: An ZNFs/LINC00520/miR-181d/BCL2 axis as a novel network in DDP-resistant LUAD has been constructed successfully, which may provide potential therapeutic targets for LUAD.

Analysis of the Long-term Effect of Intraoperative Radiotherapy (IORT) for Non-Small Cell Lung Carcinoma (NSCLC)

OBJECTIVE To analyze the long-term effects of treatment with an op-eration + postoperative irradiation (A group) and an operation+intraoperative radiotherapy+postoperative irradiation (B group) in non-small cell lung cancer patients. METHODS Through a prospective randomized clinical trial, a total of 154 patients with non-small cell lung carcinoma were divided into two groups of 77 cases. Among the 154 cases, there were 134 squamous carcinomas, 17 adenocarcinomas and 3 adeno-squamous carcinomas. TNM staging: there were 17 in StageⅠ, 76 in Stage Ⅱ and 61 in Stage Ⅲ. A dosage of 15～25 Gy IORT, energy 9～16 MeV electrons, was delivered to the tumors. The doses given were 40～60 Gy postoperation. RESULTS The local control rates in A and B groups were 49.4% and 62.3% respectively (P<0.05). The survivals at 3, 5 and 7 years for group A were 40.3%, 27.3%, and 5.2% and for group B 44.2%, 28.6% and 6.5% (P>0.05). There were 16 deaths from radiotherapy complications, with 2 cases in group A and 14 in group B. CONCLUSION IORT+postoperative irradiation can enhance the local control rate of non-small cell lung cancer patients and reduce the recurrent rates, but it can not improve long-term survival.

Relationship between the Expression of E-cadherin and Microvessel Density in Lung Cancer and Its Significance

To study the relationship between the expression of E-cadherin andmicrovessel density (MVD) in lung cancer. Methods: The expression of E-cadherin and factor VIII wasdetected in 104 lung cancer tissues by an immunohistochemical method, and MVD was calculated by animage-analysis system. Results: The expression of E-cadherin was significantly related to thedifferentiation of lung cancers (P 【 0.05). A negative correlation was found between E-cadherinexpression and MVD in lung cancer tissues (P=0.047). Conclusion: Down-expression of E-cadherin andan increase of MVD may play an important role in the invasion and metastasis of lung cancer, and mayalso be used as a useful marker for tumor prognosis.

THE CLINICAL COURSE AND TREATMENT RESULTS OF LUNG METASTASES FROM BREAST CANCER

Objective: To analyze the clinical course and treatment result of lung metastases from breast cancer Method: 122 cases with lung metastases from breast cancer were treated with chemotherapy or chemotherapy plus endocrine therapy, response was assessed according to WHO criteria and survival rate estimated using the life Table Results: The median time from initial treatment of primary tumor to lung metastases was 22 months Sites of common consecutive metastases were lung, liver and bone The overall response rate was 48% with a CR rate of 15% Compared to non DDP encompassing regimen, the CR rate was higher in DDP based chemotherapy (7% versus 21%, P <0 05) with a longer median survival time (MST) The PR rate was higher in regimens containing anthracycline (48%) than in those without anthracycline (20%, P <0 01) The response rate was similar between chemotherapy and chemotherapy plus endocrine therapy ( P >0 05) No difference in MST was observed between patients receiving anthracycline and non anthracycline encompassing regimens The 1 , 3 , 5 , and 10 year survival rate was 77%, 22%, 11%, and 10%, respectively Conclusion: Size of primary tumor, the length of disease free interval, the number of lung metastases may provide additional information for predicting patients survival after treatment of lung metastases Combination chemotherapy, especially DDP based chemotherapy may prolong survival time of patients with lung metastases from breast cancer

The Inhibitory Effects of Rh-endostatin(YH-16) in Combination with Radiotherapy on Lung Adenocarcinoma A549 in Mice and the Underlying Mechanisms

In order to investigate the inhibitory effects of Endostar(rh-endostatin,YH-16)in combination with radiotherapy on lung adenocarcinoma A549 in mice and the interaction mechanisms of combined therapy,the transplantation tumor models of A549 lung adenocarcinoma were established.When the largest diameter of tumor reached 1.0cm,all nude mice were randomly divided into 4 groups:Endostar group,radiotherapy group,radiotherapy plus Endostar(combined treatment)group,and control group(n=6 in each group).The largest d...

Antineoplastic effects of deoxyelephantopin,a sesquiterpene lactone from Elephantopusscaber, on lung adenocarcinoma (A549) cells

OBJECTIVE： Deoxyelephantopin, a sesquiterpene lactone from Elephantopus scaber, showed inhibition of the growth of various tumor cells in vitro. In the present study, we investigated the cytotoxicity and apoptosis-inducing capacity of deoxyelephantopin on lung adenocarcinoma （A549） cells. METHODS： The cytotoxic effect of deoxyelephantopin on A549 cells and normal lymphocytes was evaluated using 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide （MTT） assay and 50% inhibitory concentration （IC50） value was determined. The self-renewal and proliferating potential of A549 cells after treatment with deoxyelephantopin were examined by colony formation assay. Cellular morphology of deoxyelephantopin-treated cells was observed using phase- contrast microscopy. The induction of apoptosis was evaluated using acddine orange and ethidium bromide staining, Hoechst 33342 staining, terminal deoxynucleotidyl transferase-mediated dUTP biotin nick end-labeling （TUNEL） assay, DNA fragmentation analysis and Annexin V-fluorescein isothiocyanate staining by flow cytometry. Activation of caspases was detected using fluorogenic substrate specific to caspases 2, 3, 8 and 9 and flow cytometric analysis. The total cellular DNA content and expression of cleaved poly （ADP-ribose） polymerase was also analyzed. RESULTS： Deoxyelephantopin exhibited cytotoxicity to A549 cells （IC50 = 12.287 μg/mL）, however, there was no toxicity towards normal human lymphocytes. Deoxyelephantopin suppressed the colony-forming ability of A549 cells in a dose-dependent manner. Acridine orange, ethidium bromide and Hoechst 33342 staining showed cell shrinkage, chromosomal condensation and nuclear fragmentation, indicating induction of apoptosis. Deoxyelephantopin increased apoptosis of A549 cells, as evidenced by more TUNEL-positive cells. DNA fragmentation and Annexin V staining revealed late-stage apoptotic cell population. Deoxyelephantopin inhibited A549 cell growth by cell cycle arrest at G2/M phase and induced apoptosis through both extrinsic and intrinsic pathways. CONCLUSION： These results suggest that deoxyelephantopin has great potential as a new chemotherapeutic agent to be developed further for the treatment of lung cancer.

Helicobacter pyloriseroprevalence in patients with lung cancer

AIM:To assess Helicobacter pylori(Hpylon)seroprevalence in a cohort of Greek patients with lung cancer. METHODS:Seventy-two lung cancer patients(55 males and 17 females,aged 58.2±11.7 years)and 68,age and gender-matched,control subjects were enrolled.All subjects underwent an enzyme-linked immunosorbent assay IgG serologic test for Hpylori diagnosis. RESULTS:A correlation between age and HpyloriIgG level was detected for both lung cancer patients(r=0.42, P=0.004)and controls(r=0.44,P=0.004).Seropositivity for Hpyloridid not differ significantly between patients with lung cancer and controls(61.1% vs 55.9%,P>0.05). Concerning the mean serum concentration of IgG antibodies against Hpylori,no significant difference between the two groups was detected(32.6±19.1 vs 27.4±18.3 U/mL, P>0.05). CONCLUSION:No significant association between Hpylori infection and lung cancer was found.

DIAGNOSTIC VALUE OF SERUM PROGRP31-98 IN PATIENTS WITH SMALL-CELL LUNG CANCER

Objective To explore the clinical significance of serum level of pro gastrin releasing peptide 31 98 (ProGRP31 98) for small cell lung cancer (SCLC), in comparison with neuron specific enolase (NSE). Methods Serum level of ProGRP31 98 and NSE was measured by ELISA respectively in 30 patients with SCLC, 30 with non small cell lung cancer (NSCLC), 15 with benign lung diseases and 15 normal subjects, additionally, 10 SCLC patients after having treatment with chemotherapy were included. The receiver operating characteristic (ROC) curve was used to set the cut off value and evaluate the diagnostic accuracy. Results The serum level of ProGRP31 98 was higher in patients with SCLC than that in other groups. The SCLC patients with extensive disease had a higher value than the patients with limited disease. In SCLC patients with distant metastases, it was also higher than in those without. Increase in serum ProGRP31 98 and NSE was both seen in SCLC patients, but for the former one, the increase was of much greater compared to the normal controls. Given the cut off value for ProGRP31 98 was 40ng·L -1 and for NSE 8μg·L -1 , their sensitivity of diagnosis in SCLC was 73% and 60%, respectively. The area under ROC curve of ProGRP31 98 was significantly larger than that of NSE. All patients responded to chemotherapy showed marked decrease in ProGRP31 98. Conclusion ProGRP31 98 is a more specific and sensitive marker than NSE in the diagnosis of SCLC.

Health beliefs toward lung cancer screening among Chinese American high-risk smokers:Interviews based on Health Belief Model

Objective This study aims to explore health beliefs toward lung cancer screening with low dose computed tomography among Chinese American high-risk smokers.Methods Guided by the Health Belief Model,semi-structured individual interviews were conducted with Chinese American high-risk smokers via phone.Additional questionnaires on demographic information,history of smoking and lung cancer screening were collected via email or phone before the interview,depending on participants’preference.Content analysis was used to extract meaningful and significant themes in the dataset.Constant comparison analysis and process coding were used to categorize and code data.Results Data saturation was reached after interviewing 12 participants.Chinese American high-risk smokers perceived a low susceptibility to lung cancer,since they believed various protective factors of lung cancer(e.g.,doing exercise,healthy diet,etc.)reduced their risk of getting lung cancer.All the participants perceived a high severity of lung cancer.They acknowledged lung cancer would have a huge impact on their life.Perceived benefits of lung cancer screening were accurate in most aspects although minor confusions were still noticed among this population.Perceived barriers varied on participants’,physicians’,and institutional levels.High-risk Chinese American smokers had little confidence to screening for lung cancer.Cues to action for them to screening for lung cancer included recommendations from health care providers,support from family members and friends,and information shared on Chinese-based social media.Conclusions Misconceptions and barriers to screening for lung cancer existed widely among Chinese American high-risk smokers.Intervention programs and targeted health education should be implemented to promote lung cancer screening among this population.

A Controlled Study on Comparing Differences in CT Perfusion Imaging between Rabbits inoculated with VX2 Lung Tumor and Patients with Squamous Cell Carcinoma of the Lung

OBJECTIVE By analysis and evaluation of the perfusion images and perfusion parameters of the rabbits with VX2 lung tumor, the association between the perfusion parameters and tumor angiogenesis of patients with squamous cell carcinoma of the lung has been studied in order to establish a non-invasive and effective way to detect tumor blood supply, which is be able to exhibit hemodynamic data in tumors during cancer treatments. METHODS Fifteen Netherlands rabbits inoculated with VX2 lung tumor （rabbit group） and 25 patients with squamous cell carcinoma of the lung （patient group） received a multi-slice spiral CT perfusion imaging test using the Netherlands PHILIPS Brilliance 16-slice spiral CT and a U.S. MEDRAD binocular highpressure syringe. Image postprocessing was done using the special perfusion software and EBW 4.0 Workstation. Perfusion volume （PV）, peak enhanced increment （PEI）, transit time peak （TTP）, and blood volume （BV） were measured and analyzed. RESULTS In the rabbit group, the values of the PV, PEI, TTP, and BV of the tumor margin were （53.89 ± 13.38） mL/（min.mL）, （45.71 ± 15.52） Hu, （39.29 ± 10.10） sec, and （31.45 ± 18.19） mL/100 g, respectively; these values of the tumor center were （36.57 ± 14.17） mL/（min.mL）, （28.64 ± 11.74） Hu, （39.00 ＋ 9.78） sec, and （19.76 ± 13.95） mL/100 g, respectively; the values of the muscles were （12.45± 4.38） mL/（min.mL）, （10.98 ± 5.03） Hu, （38.86 ± 10.04） sec, and （5.38 ±2.87） mL/100 g, respectively. The values of the relative perfusion volume （RPV）, relative peak enhanced increment （RPEI）, and relative blood volume （RBV） of the tumor margin were 4.38 ± 1.45, 3.96± 1.45, 9.99 ± 11.7, respectively; these values of the tumor center were 2.14 ± 1.08, 1.83±1.45, 4.17 ±3.39, respectively. The values of the PV, PEL BV of the tumor margin vs. the values of the muscles developed t-values, which were 15.028, 10.79, and 5.88, respectively （P ≤ 0.01）, with statistical significance; the values of the PV, PEI, BV of the tumor center vs. the values of the muscles produced t-values, which were 8.67, 7.49, and 4.55, respectively （P 〈 0.01）, with statistical significance. The values of the TTP of the tumor margin vs. TTP values of the muscles, and the TTP values of the tumor center vs. TTP values of the muscles developed t-values, which were 1.7 and 0.806, respectively （P ≥ 0.05）, without statistical significance. In the patient group, the values of the PV, PE, TTP, and BV of the tumor margin were （88.95 ± 30.89） mL/（min.mL）, （61.87 ± 27.31） Hu, （37.72 ± 12.53） sec, and （18.38 ± 7.2） mL/100 g, respectively; these values of the tumor center were （39.77 ± 18.29） mL/（min.mL）, （14.57 ± 8.1） Hu, （35.64 ± 12.41） sec, and （11.22 ± 6.02） mL/100 g, respectively; these values of the muscles were （12.45 ± 6.5） mL/（min.mL）, （6.14 ± 2.66） Hu, （35.68± 12.35） sec, and （2.23 ± 1.11） mL/100 g, respectively. The values of the RPV, RPEI, and RBV of the tumor margin were 8.05 ± 5.04, 8.87 ± 4.32, and 12.16 ± 8.49, respectively; these values of the tumor center were 2.39 ± 1.68, 2.97 ± 2.1, 3.53 ± 2.82, respectively. The values of the PV, PEI, BV of the tumor margin in the patient group vs. the values of the muscles produced t-values, which were 13.8, 10.85, and 12.22, respectively （P 〈 0.01）, with significant differences; these values of the tumor center vs. the values of the muscles developed t-values, which were 9.158, 6.26, 8.654, respectively （P 〈 0.01）, with significant differences. The TTP value of the tumor margin vs. that of the muscles produced t-value, which was 0.371, and the TTP value of the tumor center vs. that of the muscles developed t-value, which was 1 （P 〉 0.05）, without statistical difference. CONCLUSION CT perfusion imaging technics demonstrates directly dynamic changes of blood flow to tumors, which assists in identifying tumor growth and necrosis, therefore, this research provides an evidence-based guidelines for the treatment of human lung squamous cell carcinoma and has far-reaching clinical significance.

The effects of CYP1A1 gene polymorphism and p16 gene methylation on the risk of lung cancer in a Chinese population

Objective： To investigate the relationship between the genetic polymorphism of CYP1A1 and the genetic susceptibility to lung cancer as well as to study the effects of the methyiation in p16 gene on the risk of lung cancer in a Chinese population. Methods： A case control study was conducted among 47 cases of lung cancer and 94 controls. The genetic polymorphism of CYP1A1 was tested with method of PCR-RFLP, and a methylation-specific PCR （MSP） was performed to detect p16 methylation. Results： It showed that there was no significant difference in frequencies of the genotypes of CYP1A1 between the two groups （P 〉 0.05）. Synergistic effects were not found between smoking and CYP1AI. Methylated p16 gene was found in 44.7% （21/47） of lung cancer tissues and in 17.0% （8/47） of normal lung tissues with significant difference （P 〈 0.05）. Conclusion： The genetic polymorphism of CYP1A1 does not increase the risk of lung cancer in a Chinese population. The methylation in p16 gene may be the most common mechanism to inactivate p16 gene in lung cancer, and is not significantly associated with genotype of CYP1A1,

Phase Ⅰ/Ⅱ study of gemcitabine and oxaliplatin chemotherapy in combination with concurrent 3-D conformal radiotherapy for locally advanced non-small cell lung cancer

Background and objective Recent studies have showed that combination of chemotherapy and radiotherapy might result in better outcome for locally advanced non-small cell lung cancer (NSCLC). The aim of this study is to determine the maximal tolerance dose (MTD) and efficacy of full-dose gemcitabine and oxaliplatin when given concurrently with 3-dimentional radiation therapy (3D-RT) for locally advanced NSCLC. Methods Oxaliplatin was administered at a fixed dose of 130mg/m^2, and gemcitabine was administered at a starting dose of 800mg/m^2 with an incremental dose gradient of 200mg/m^2 for 3 dose levels. MTD was defined as the immediate dose level lower than the dose at which dose-limiting toxicity (DLT) occurred in more than one-third of the patients. The chemotherapy was administered at 3-week cycle. The RT was given as 3-D conformal manner at a single daily dose of 2Gy for 5 days per week. Results Twenty-two patients were evaluable and distributed to three different dose levels: 6 at level 1, 8 at level 2 and 8 at level 3. Pulmonary toxicity, esophageal and hematologic toxicity were the main DLT. Grade Ⅲ acute pulmonary toxicity occurred in one patient each at level 2 and level 3, both with V20>20%, and grade Ⅲ esophagitis in two patients at level 3. The MTD of gemcitabine in this study was 1000mg/m^2. The overall response rate was 75.0% (9/12). The 1- and 2-year survival rate was 70.0% and 30.5% respectively. The median time to progression was 8.7 months (range 5--11.8 months). Conclusion With reduced radiation volume, gemcitabine of 1000mg/m^2 in combination with oxaliplatin of 130mg/m^2 was effective and could be safely administered for NSCLC.

Treatment Sequencing Strategies in Lung Cancer

Background and objective The advances in the lung cancer screening methods and therapeutics,together with awareness towards deleterious habits,such as smoking,is increasing the overall survival with better quality of life for the patients.However,lung cancer is still one of the most common and fatal neoplasm with a high incidence and consequently burden to public health worldwide.Thus,based on guidelines and recent phasesⅡandⅢclinical trials studies,this manuscript summarizes the current treatment sequencing strategies in lung cancer.Methods A comprehensive search of related articles was performed focused on phasesⅡandⅢclinical trials studies.Results The lung cancer management should take into consideration the tumor characteristics,histology,molecular pathology and be discussed in a multidisciplinary team.Lung cancer treatment options comprises surgery whenever possible,radiotherapy associate with/or chemotherapy and immunotherapy as monotherapy,or combined with chemotherapy and best palliative care.Conclusions The screening predictability in more patients,smoking reduction,early diagnosis,better disease understanding and individualized,more effective and tolerable therapeutics are related to an increasing in overall survival and quality of life.In the near future improvement of personalized therapy in precision medicine is expected,enhancing new predictive biomarkers,optimal doses and optimal treatment sequencing as well as anti-cancer vaccines development.

Paraneoplastic Leukocytosis and Thrombocytosis as Prognostic Biomarkers in Non-small Cell Lung Cancer

Background and Objectives: Search for inexpensive laboratory markers have identified associations between blood counts and lung cancer outcomes. In this study, we evaluated the prognostic value of paraneoplastic leukocytosis(p-Leukocytosis) and paraneoplastic thrombocytosis(p-Thrombocytosis) in patients with non-small cell lung cancer(NSCLC). We also studied their relation to the expression of commonly detected molecular markers. Methods: We conducted a retrospective chart review on 571 consecutive NSCLC patients over a 10 year period. Blood counts were recorded at the time of cancer diagnosis. Kaplan-Meier survival curves were used to compare overall survival(OS) between patients with and without p-Leukocytosis(or) p-Thrombocytosis(p-Leuko/Thrombocytosis). Cox regression was used to determine if leukocytosis/thrombocytosis was a predictor of OS in NSCLC.Results: Patients with p-Leukocytosis and p-Thrombocytosis had a significantly poorer survival compared patients with normal blood counts(P<0.001). In a multivariate survival analysis, both continued to correlate even when adjusted for histology, gender, stage and chemotherapy(P<0.01, 0.03 respectively). Stage I and II NSCLC with p-Leuko/Thrombocytosis did not perform poorly compared to stage I/II NSCLC patients without paraneoplasia. Patients with the combined leukothrombocytosis syndrome did not have worse outcomes compared to those with either paraneoplastic syndrome alone. Conclusions: p-Leuko/Thrombocytosis is an accessible laboratory parameter of prognostic value in NSCLC. Evidence of p-Leuko/Thrombocytosis portends poor survival. The role of various cytokines in tumor pathobiology provides a rationale for identifying cytokine factors responsible for the paraneoplasia and administering anti-cytokine therapies alongside traditional chemotherapy in an attempt to improve survival outcomes in these subset of patients.

Establishment and application of a multiplex genetic mutation-detection method of lung cancer based on MassARRAY platform

Objective: This study aims to establish a method for highly parallel multiplexed detection of genetic mutations in Chinese lung cancer samples through Agena i PLEX chemistry and matrix-assisted laser desorption ionization time-of-flight analysis on Mass ARRAY mass spectrometry platform.Methods: We reviewed the related literature and data on lung cancer treatments. We also identified 99 mutation hot spots in 13 target genes closely related to the pathogenesis, drug resistance, and metastasis of lung cancer. A total of 297 primers, composed of99 paired forward and reverse amplification primers and 99 matched extension primers, were designed using Assay Design software. The detection method was established by analyzing eight cell lines and six lung cancer specimens. The proposed method was then validated through comparisons by using a Lung Carta^(TM) kit. The sensitivity and specificity of the proposed method were evaluated by directly sequencing EGFR and KRAS genes in 100 lung cancer cases.Results: The proposed method was able to detect multiplex genetic mutations in lung cancer cell lines. This finding was consistent with the observations on previously reported mutations. The proposed method can also detect such mutations in clinical lung cancer specimens. This result was consistent with the observations with Lung Carta^(TM) kit. However, an FGFR2 mutation was detected only through the proposed method. The measured sensitivity and specificity were 100% and 96.3%, respectively.Conclusions: The proposed Mass ARRAY technology-based multiplex method can detect genetic mutations in Chinese lung cancer patients. Therefore, the proposed method can be applied to detect mutations in other cancer tissues.

Efficacy Differences of First-line EGFR-TKIs Alone vs in Combination with Chemotherapy in Advanced Lung Adenocarcinoma Patients with Sensitive EGFR Mutation and Concomitant Non-EGFR Genetic Alterations

Background and objective:Epidermal growth factor receptor(EGFR)mutations are often associated with non-EGFR genetic alterations,which maybe a reason for the poor efficacy of EGFR tyrosine kinase inhibitors(TKIs).Here we conducted this study to explore whether EGFR-TKIs combined with chemotherapy would benefit advanced lung adenocarcinoma patients with both sensitive EGFR mutation and concomitant non-EGFR genetic alterations.Materials and methods:Cases of advanced lung adenocarcinoma with EGFR mutation combined with concomitant nonEGFR genetic alterations were retrospectively collected.And the patients were required to receive first-line EGFR-TKIs and chemotherapy combination or EGFR-TKIs monotherapy.Demographic,clinical and pathological data were collected,and the electronic imaging data were retrieved to evaluate the efficacy and time of disease progression.Survival data were obtained through face-to-face or telephone follow-up.The differences between the two groups in objective response rate(ORR),disease control rate(DCR),progression-free survival(PFS)and overall survival(OS)were investigated.Results:107 patients were included,including 63 cases in the combination group and 44 cases in the monotherapy group.The ORR were 78%and 50%(P=0.003),and DCR were 97%and 77%(P=0.002),respectively.At a median follow-up of 13.7 mon,a PFS event occurred in 38.1%and 81.8%of patients in the two groups,with median PFS of18.8 mon and 5.3 mon,respectively(P<0.000,1).Median OS was unreached in the combination group,and 27.8 mon in the monotherapy group(P=0.31).According to the Cox multivariate regression analysis,combination therapy was an independent prognostic factor of PFS.Conclusion:In patients with EGFR-mutant advanced lung adenocarcinoma with concomitant non-EGFR genetic alterations,combination of TKIs and chemotherapy was significantly superior to EGFR-TKIs monotherapy,which should be the preferred treatment option.

Inhalation treatment of lung cancer: the influence of composition, size and shape of nanocarriers on their lung accumulation and retention

Objective： Various nanoparticles have been designed and tested in order to select optimal carriers for the inhalation delivery ofanticancer drugs to the lungs. Methods： q-he following nanocarriers were studied： micelles, liposomes, mesoporous silica nanoparticles （MSNs）, poly propyleneimine （PPI） dendrimer-siRNA complexes nanoparticles, quantum dots （QDs）, and poly （ethylene glycol） polymers. All particles were characterized using the following methods： dynamic light scattering, zeta potential, atomic force microscopy, in vitro cyto- and genotoxicity. In vivo organ distribution of all nanopartides, retention in the lungs, and anticancer effects of liposomes loaded with doxorubicin were examined in nude mice after the pulmonary or intravenous delivery. Results： Significant differences in lung uptake were found after the inhalation delivery of lipid-based and non-lipid-based nanoparticles. The accumulation ofliposomes and miceUes in lungs remained relatively high even 24 h after inhalation when compared with MSNs, Q Ds, and PPI dendrimers. There were notable differences between nanoparticle accumulation in the lungs and other organs 1 and 3 h after inhalation or intravenous administrations, but 24 h after intravenous injection all nanoparticles were mainly accumulated in the liver, kidneys, and spleen. Inhalation delivery of doxorubicin by liposomes significantly enhanced its anticancer effect and prevented severe adverse side effects of the treatment in mice bearing the orthotopic model of lung cancer. Conclusion： The results of the study demonstrate that lipid-based nanocarriers had considerably higher accumulation and longer retention time in the lungs when compared with non-lipid-based carriers after the inhalation delivery. These particles are most suitable for effective inhalation treatment of lung cancer.

HYPERMETHYLATION OF p14^(ARF) PROMOTER REGION AND EXPRESION OF p14^(ARF) GENE PRODUCT IN NON-SMALL CELL LUNG CANCER

Objective： This study was designed to investigate promoter methylation status and protein expression of p14^ARF gene in non-small cell lung cancer, and value the role of p14^ARF promoter methylation in carcinogenesis of non-small cell lung cancer. Methods： Promoter methylation status and protein expression of p14^ARF gene in 40 cases of non-small cell lung cancer were analyzed by methylation specific polymerase china reaction （MSP）, restriction enzyme-related polymerase chain reaction （RE-PCR） and immunohistochemistry （IHC）. Results： The positive rates of p14^ARF promoter methylation in tumor tissues and normal tissues adjacent to cancer were 17.5% （7/40） and 2.5% （1/40） respectively. There were statistically significant differences between them, P〈0.05. The results of RE-PCR were consistent with that of MSP. The expression rate of p14^ARF protein in tumor tissues was significantly lower than that in normal tissues adjacent to cancer, p〈0.01. Promoter methylation status and protein expression of p14^ARF gene in non-small cell lung cancer showed significantly an inverse correlation （r=-0.56, P〈0.01）, and both of them did not relate statistically with the clinicopathologic characteristics of patients such as histological classification, clinical stage, differentiation grade and lymph node involvement. Conclusion： Promoter methylation is a crucial mechanism of inactivation of p14^ARF gene. Promoter methylation of p14^ARF gene might he involved in carcinogenesis of non-small cell lung cancer, and is an early event in development process of non-small cell lung cancer. It might be used as a new target in gene treatments in the future.

CTLA-4+49 A/G Polymorphism and the Risk of Lung Cancer:a Meta-analysis

Background and objective:Lung cancer is one of the malignant tumors.Gene mutations associated with cellular immune function and regulating the activation and proliferation of immune cells.Several publications have explored the relationship between cytotoxic T lymphocyte antigen-4(CTLA-4)+49 adenine(A)/guanine(G)polymorphism and susceptibility of lung cancer,but the results remain controversial.Thus,we performed this meta-analysis to derive a more comprehensive estimation of the relationship.Methods:All articles addressed lung cancer and polymorphisms of CTLA-4 were searched from the Pub Med,EMBASE databases published up to June 29,2019.Odds ratios(ORs)with 95%confidence intervals(CIs)were used to assess the strength of association.Publication bias of relevant studies was examined via Begg’s test and funnel plots.Results:The meta-analysis included 8 case-control studies covering 4,430 lung cancer patients and 5,198 healthy controls from September 2008 to April 2020.The overall eligible data indicated that CTLA-4+49 A/G polymorphisms did not correlate with the elevated lung cancer risk in all genetic comparison models(dominant model:OR=1.037,95%CI:0.925-1.161;recessive model:OR=0.968,95%CI:0.888-1.055;allele model:OR=0.992,95%CI:0.933-1.054;homozygous model:OR=0.980,95%CI:0.857-1.121;heterozygous model:OR=1.023,95%CI:0.906-1.154).In further stratified analyses,CTLA-4+49 A/G polymorphism was found to be significantly associated with susceptibility to NSCLC in these models(dominant model:OR=1.404,95%CI:1.074-1.836;allele model:OR=1.273,95%CI:1.034-1.565;homozygous model:OR=1.553,95%CI:1.044-2.310;heterozygous model:OR=1.308,95%CI:1.062-1.611).Conclusion:CTLA-4+49 A/G polymorphism were not associated with the risk of lung cancer but might be a risk factor only in NSCLC.