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We reported unique molecular features of cerebrospinal fluid (CSF) of nonsmall cell lung cancer (NSCLC) patients with leptomeningeal metastasis (LM), suggesting establishing CSF as a better liquid biopsy in clinical practices. We performed next-generation panel sequencing of primary tumor tissue, plasma, and CSF from 131 NSCLC patients with LM and observed high somatic copy number variations (CNV) in CSF of NSCLC patients with LM. The status of EGFR-activating mutations was highly concordant between CSF, plasma, and primary tumors. ALK translocation was detected in 8.3% of tumor tissues but only 2.4% in CSF and 2.7% in plasma. Others such as ROS1 rearrangement, RET fusion, HER2 mutation, NTRK1 fusion, and BRAF V600E mutation were detected in 7.9% of CSF and 11.1% of tumor tissues but only 4% in plasma. Our study has shed light on the unique genomic variations of CSF and demonstrated that CSF might represent better liquid biopsy for NSCLC patients with LM.

QTL mapping for fiber-related traits and elucidation of a stable and novel QTL affecting seed lignin content, cellulose content and seed oil content. Dissection of the genetic networks for fiber biosynthesis is important for improving the seed oil content and meal value of Brassica napus. In this study, the genetic basis of seed fiber biosynthesis in B. napus was investigated via quantitative trait locus (QTL) analysis using a doubled haploid population derived from ‘KenC-8’ crossed with ‘N53-2.’ Seed lignin content (LC), cellulose content (CC) and hemicellulose content (HC) were significantly negatively correlated with seed oil content (OC). Co-localization QTLs among LC, CC, HC and OC on A09 were found with contributions ranging from 9.87 to 48.50%. Seven co-localization QTLs involved in the fiber component and OC were further verified by bulked segregant analysis (BSA). The unique QTL uqA9-12 might be a real and new QTL that was commonly identified by QTL mapping and BSA and simultaneously affected LC, CC and OC with opposite additive effects. A potential regulatory network controlling seed fiber biosynthesis was constructed to dissect the complex mechanism of seed fiber and oil accumulation, and numerous candidate genes were identified in the fiber-related QTL regions. These results provided an enrichment of QTLs and potential candidates for fiber biosynthesis, as well as useful new information for understanding the complex genetic mechanism underlying rapeseed seed fiber accumulation.

9102 Background: Mefatinib is a novel, second-generation, irreversible epidermal growth factor receptor (EGFR)-tyrosine kinase inhibitor (TKI). A Phase Ib/II study has showed promising efficacy in patients with advanced EGFR (19del/L858R) mutant non-small cell lung cancer (NSCLC). This study is aimed to assess the efficacy and safety of mefatinib as first-line therapy for advanced rare EGFR mutations (G719X/S768I/L861Q) NSCLC. Methods: In this phase II study, we enrolled patients with stage IIIB-IV lung adenocarcinoma with at least one mutation in G719X, L861Q, and S768I who hadn¡¯t received front-line anticancer therapy. Other concurrent EGFR mutations were allowed with the exception of Ex19del, L858R, and exon 20 insertions. Eligible patients were treated with mefatinib 80 mg orally once daily. The primary endpoint was objective response rate (ORR) according to RECIST 1.1 criteria. Results: A total of 21 patients (median age 57.6, 47.62% male) have been enrolled, 80.95% with G719X, 33.3% S768I, and 19.05% L861Q. Six (28.57%) patients had concurrent mutations of G719X and S768I, and one (4.76%) patient had co-mutations of G719X and L861Q. As of data cut-off (Jan, 6, 2022), median follow-up was 25.1 months. Among the 21 efficacy evaluable patients, ORR was 85.71% (95% CI, 63.66-96.95) with a duration of response (DOR) of 22.2 months (95% CI, 8.4-NA). Disease control rate (DCR) was 100% (95% CI, 83.89-100.00). Median progression-free survival (mPFS) was 20.6 months (95% CI, 8.3-NA). Median overall survival (OS) was not reached.. The most common treatment-related adverse events (TRAEs) were rash (20, 95.24%), diarrhea (19, 90.48%), stomatitis (15, 71.43%), and pandactilitis (9, 42.86%). Grade 3 TRAEs were reported in 47.63% patients. Common Grade 3 TRAEs (¡Ý5%) included diarrhea (8, 38.10%) and rash (5, 23.81%). No ¡ÝGrade 4 TRAE or treatment-related SAE reported. 33.33% (n=7) of patients experienced dose reduction due to AEs (mainly diarrhea and rash). All TRAEs were reversible and remained stable or recovered without sequelae after dose reduction. None of the patients discontinued treatment or died due to TRAE. Conclusions: Mefatinib has shown promising anti-tumor activity and a favorable safety profile in patients with advanced NSCLC harboring rare EGFR mutations. Clinical trial information: CTR20190110. [Table: see text]

Background: To date, the reactogenicity and immunogenicity of heterologous ChAdOx1-nCoV19/BNT162b2 (ChAd/BNT) vaccination remain unclear. Here, we systematically assessed the reactogenicity and immunogenicity of the heterologous ChAd/BNT vaccination regimens. Methods: The publicly available studies on heterologous COVID-19 vaccination were systematically searched on PubMed, Web of Science, Embase and MedRxiv for the available literature up to August 1, 2021. We evaluated the immunogenicity by the geometric mean titers (GMTs) of the neutralizing antibody and anti-spike IgG. The reactogenicity was evaluated using the pooled risk ratios (RRs) calculated by the random-effects model about the adverse events after vaccination. Subgroup analyses based on vaccination regimens and interval between the two doses of vaccines were conducted to evaluate the impact of the different regimens. Findings: Nine studies (n=4,286) were included in the analyses. Compared to the homologous ChAd/ChAd vaccination, the heterologous ChAd/BNT vaccination showed significantly higher immunogenicity (pooled GMTR [geometric mean titers ratio] in terms of the neutralizing antibody: 11·92 [95% CI 5·65-25·13, I2=49·1%, P

Endophytic fungi play an important role in plant growth. The composition and structure of endophytes vary in different plant tissues, which are specific habitats for endophyte colonization. To analyze the diversity and structural composition of endophytic fungi from toothed clubmoss (Huperzia serrata) that was artificially cultivated for 3 years, we investigated endophytic fungi from the roots, stems and leaves using comparative sequence analysis of the ITS2 region of the fungal rRNA genes sequenced with high-throughput sequencing technology. Seven fungal phyla were identified, and fungal diversity and structure varied across different tissues, with the most distinctive community features found in the roots. A total of 555 operational taxonomic units (OTUs) were detected, and 198 were common to all samples, and 43, 16, 16 OTUs were unique to the root, stem, leaf samples, respectively. Taxonomic classification showed that Ascomycota and Basidiomycota were dominant phyla, and Cladosporium, Oidiodendron, Phyllosticta, Sebacina and Ilyonectria were dominant genera. The relative abundance heat map at the genus level suggested that H. serrata had characteristic endophytic fungal microbiomes. Line discriminant analysis effect size analysis and principal coordinate analysis demonstrated that fungal communities were tissue-type and tissue-site specific. Overall, our study provides new insights into the complex composition of endophytic fungi in H. serrata.

Genome-wide chromatin reprogramming is a fundamental requirement for establishing developmental competence in the newly-formed zygote. In zebrafish, Nanog, Pou5f3 and Sox19b play partially redundant roles in zygotic genome activation, however their interplay in establishing chromatin competency, the context in which they do so and their mechanism of action remain poorly defined. Here, we generated a triple maternal-zygoticnanog-/-;pou5f3-/-;sox19b-/-mutant and assessed the causal relationship between transcription factor (TF) occupancy, chromatin accessibility and genome activation. Analyses of this triple mutant and combinatorial rescues revealed highly synergistic and context-dependent activity of Nanog, Pou5f3, and Sox19b (NPS) in establishing chromatin competency at >50% of active enhancers. Motif analysis revealed a network of TFs that depend on NPS for establishing chromatin accessibility, including the endodermal determinant Eomesa, whose binding we show is regulated by NPS pioneer-like activity. Finally, we demonstrated that NPS play an essential role in establishing H3K27ac and H3K18ac at enhancers and promoters, and that their function in transcriptional activation can be bypassed by targeted recruitment of histone acetyltransferases to individual genes. Altogether, our findings reveal a large network of TFs that function to establish developmental competency, many of which depend on the synergistic and highly context-dependent role of NPS in establishing chromatin accessibility and regulating histone acetylation in order to activate the genome.

To mine differentially expressed microRNAs (miRs) in lung adenocarcinoma (LUAD) via The Cancer Genome Atlas (TCGA).The transcriptome and pathological data of LUAD patients were downloaded from TCGA. The differentially expressed genes were screened using "edgeR" package in R and analyzed by univariate and multivariate Cox regressions. The expression and clinic value of serum miR-548v in 50 patients with LUAD (study group) and 50 healthy individuals (control group) was detected by the quantitative real time polymerase chain reaction (qRT-PCR), and the diagnostic value of miR-548v in LUAD was analyzed by the receiver operating characteristic (ROC) curve.A total of 233 differentially expressed genes were found, of which 115 were highly expressed and 118 were lowly expressed. Multivariate Cox regression showed that hsa-mir-142 and hsa-mir-548v were independent prognostic factors for patients with LUAD. The study group showed significantly higher serum miR-548v expression than the control group (p0.001). miR-548v was related to TNM staging, lymph node metastasis, and tumor size of patients (p0.05). The area under the curve (AUC) of miR-548v in diagnosing LUAD was 0.960, and that in determining TNM staging, lymph node metastasis and tumor size was 0.829, 0.851 and 0.881 respectively.Differential expression of miR-548v predicts poor prognosis of patients with LUAD and is closely related to TNM staging, lymph node metastasis and tumor size. Therefore, it is expected to be a potential diagnostic and prognostic marker for LUAD.

Stress conditions induced by routine treatments might affect cancer-associated fibroblasts in lung adenocarcinoma. The present study tried to explore transcriptome changes in lung fibroblasts under chemotherapeutics, irradiation, and hypoxia, which were induced by chemotherapy, radiotherapy, and anti-angiogenesis therapy, respectively. We established three in vitro models to mimic the stress conditions for lung fibroblasts. Interestingly, one of the secretory molecules, tumor necrosis factor superfamily member 4 (TNFSF4, also known as OX40L), was significantly up-regulated in lung fibroblasts under stress environments. Lung adenocarcinoma patients received chemotherapy and radiotherapy had a higher expression level of TNFSF4 in serum and tumor tissues. There was a negative correlation between the increase of serum TNFSF4 levels and the shrink of the tumor after chemotherapy. TNFSF4 could promote cisplatin resistance and inhibit the apoptosis of lung adenocarcinoma cells. Furthermore, TNFSF4 could significantly increase the activity of NF-κB/BCL-XL pathway in lung adenocarcinoma cells, which could be counteracted by knocking down the expression of TNFRSF4 (receptor of TNFSF4). In conclusion, TNFSF4, secreted by cancer-associated fibroblasts under stress conditions, could facilitate chemoresistance of lung adenocarcinoma through inhibiting apoptosis of tumor cells.

Upon fertilization, embryos undergo chromatin reprogramming and genome activation, however, the mechanisms that regulate these processes are poorly understood. Here, we generated a triple mutant for Nanog, Pou5f3, and Sox19b (NPS) in zebrafish and found that NPS pioneer chromatin opening at >50% of active enhancers. NPS regulate acetylation across core histones at enhancers and promoters, and their function in gene activation can be bypassed by recruiting histone acetyltransferase to individual genes. NPS pioneer chromatin opening individually, redundantly, or additively depending on sequence context and we show that high nucleosome occupancy facilitates NPS pioneering activity. Nucleosome position varies based on the input of different transcription factors (TFs), providing a flexible platform to modulate pioneering activity. Altogether, our results illuminate the sequence of events during genome activation and offer a conceptual framework to understand how pioneer factors interpret the genome and integrate different TF inputs across cell types and developmental transitions.

A subgroup H of a finite group G is said to be weakly $$\mathcal {M}$$ -supplemented in G if there exists a subgroup B of G such that (1) $$G=HB$$ and (2) $$H_1B = BH_1

Summary The syncytiotrophoblast layer is the most critical and prominent tissue in placenta. SynT cells are differentiated from trophoblast stem cells (TSCs) during early embryogenesis. Mouse TSCs can spontaneously differentiate into cells of mixed lineages in vitro upon withdrawal of stemness-maintaining factors. However, differentiation into defined placental cell lineages remains challenging. We report here that canonical Wnt signaling activation robustly induces expression of SynT-II lineage-specific genes Gcm1 and SynB and suppresses markers of other placental lineages. In contrast to mouse TSCs, the induced SynT-II cells are migratory. More importantly, the migration depends on hepatocyte growth factor (HGF) and the c-MET signaling axis. Furthermore, HGF-expressing cells lie adjacent to SynT-II cells in developing murine placenta, suggesting that HGF/c-MET signaling plays a critical role in SynT-II cell morphogenesis during the labyrinth branching process. The availability of SynT-II cells in vitro will facilitate molecular understanding of labyrinth layer development., Highlights • Wnt is sufficient to induce SynT-II cells from trophoblast stem cells • Induced SynT-II cells are migratory and are independent on EMT • Hepatocyte growth factor/c-MET is essential for SynT-II cell migration, Zhu and colleagues successfully induce mouse syncytiotrophoblast (SynT) layer II cells from trophoblast stem cells by activation of canonical Wnt signaling. The induced SynT-II cells are migratory and are dependent on HGF/c-MET pathway. The availability of SynT-II cells in vitro should facilitate molecular study of labyrinth layer development in placenta.

A subgroup H of a finite group G is said to be S-semipermutable in G if H permutes with all Sylow q-subgroups of G for the primes q not dividing the order of H. Some criteria for p-supersolvability of a finite group are given, which are the generalizations of many recent results.

α1-antitrypsin (AAT) has been recognized to be associated with lung adenocarcinoma metastasis. However, the mechanisms by which AAT promotes tumor metastasis remain to be investigated. Herein, we first examined AAT expression in a panel of formalin-fixed paraffin-embedded tumor tissues from 88 lung adenocarcinoma patients undergoing curative resection, using immunohistochemical methods. Lung adenocarcinoma patients with high AAT expression showed a significantly shorter overall survival compared to those with low AAT expression by Kaplan-Meier method (P=0.008). High AAT expression was also identified as an independent prognostic factor by Cox regression analysis (adjusted hazard ratio: 2.05; P=0.04). Second, the role of AAT in lung adenocarcinoma cell migration was evaluated in vitro using wound healing and Transwell assays, by transfecting the lentivirus vector with interfering sequence or coding sequence of AAT. The migration property of A549 and SPC-A1 cells was significantly diminished by downregulating AAT expression. Conversely, the migration of both cell lines was significantly increased through upregulating AAT. Furthermore, AAT could increase the expression of fibronectin (FN). FN down-regulation reversed AAT-induced promotion of adenocarcinoma cell migration. Third, a cancer cell/endothelial cell co-culture model was established to investigate the effect of AAT on adenocarcinoma cell adhesion using immunofluorescence examination. The results showed that downregulation of AAT inhibited adhesion between lung adenocarcinoma cells and human umbilical vein endothelial cells whereas upregulation of AAT promoted adhesion, which may attribute to interactions between FN and integrin α5. Finally, AAT also showed the regulation effect on the metastatic behavior of lung adenocarcinoma cells in a mouse model, which may be through regulating FN expression. This study suggested that high AAT expression might be a negative prognostic marker for lung adenocarcinoma. AAT promoted lung adenocarcinoma metastasis, whose functional target may be FN. Our findings provide new insight into the mechanisms of lung adenocarcinoma metastasis.

Background: We retrospectively reported unique molecular features of cerebrospinal fluid(CSF) of NSCLC patients with LM, suggesting to establish CSF as a better liquid biopsy method in clinical practices. Methods: We performed next-generation panel sequencing of primary tumor tissue, plasma and cerebrospinal fluid (CSF) from 131 NSCLC patients with LM. Findings: High somatic copy number variations (CNV) were observed in CSF compared with plasma and primary tumor, exhibiting unique molecular features of CSF of NSCLC patients with LM. The status of EGFR-activating mutations was highly concordant between CSF, plasma, and primary tumors. There was no statistical difference between CSF, plasma and tumor tissues to detect EGFR mutation. ALK translocation was detected in 8.3% of tumor tissues, but detection rate was only 2.4% in CSF and 2.7% in plasma. Except EGFR and ALK, other rare driver genes such as ROS1 rearrangement, RET fusion, HER2 mutation, NTRK1 fusion, and BRAF V600E mutation were detected in 7.9% of CSF and 11.1% of tumor tissues, but only 4% in plasma. Except EGFR, TP53 and RB1 aberrations were high in these patients with CSF and primary tumor samples, implicating a prediction of high LM risk. Interestingly, co-occurring of EGFR and TP53 was not different among CSF, plasma and tumor tissues. However, co-occurring of EGFR and RB1 and EGFR/TP53/RB1 were higher in CSF compared with plasma or tumor tissues. Interpretation: Our study has shed light on the unique genomic variations of CSF and demonstrated that CSF may represent better liquid biopsy for NSCLC patients with LM. Funding Statement: This study was funded by the grants from the National Young 1000 Talents Program of China and Jiangsu Province Education Department, Jiangsu Province “Innovative and Entrepreneurial Team” and “Innovative and Entrepreneurial Talent.” Declaration of Interests: The authors stated: "None." Ethics Approval Statement: All patients provided signed informed consent and the study protocol was approved by the Research Ethics Committee of Nanjing Drum Tower Hospital.

The basic helix-loop-helix (bHLH) transcription factors are negatively regulated by inhibitor of DNA-binding (ID) proteins. Several studies have demonstrated that ID family proteins are dysregulated in a variety of cancer types, including in lung adenocarcinoma (LUAD). In current study, the prognostic value of ID family members was evaluated by investigating publicly accessible databases, including Oncomine, Kaplan-Meier plotter, UALCAN and the Human Protein Atlas. It was observed that the mRNA expression of all ID members was downregulated in LUAD tumor tissues compared with those in normal tissues according to the Oncomine and UALCAN databases. Additionally, increased mRNA expression levels of ID2 and ID1 were associated with improved and poorer survival time, respectively. Notably, ID3 and ID4 expression was not associated with survival in patients with LUAD. At the protein level, high ID2 significantly predicted an improved survival outcome while high ID1 is associated with shorter survival time. Thus, the results indicate that the ID proteins, particularly ID2, exhibit significant prognostic value in LUAD. More studies are required to elucidate the underlying molecular mechanisms behind the role of the ID family in the development of LUAD.

The awakening of the genome after fertilization is a cornerstone of animal development. However, the mechanisms that activate the silent genome after fertilization are poorly understood. Here, we show that transcriptional competency is regulated by Brd4- and P300-dependent histone acetylation in zebrafish. Live imaging of transcription revealed that genome activation, beginning at the miR-430 locus, is gradual and stochastic. We show that genome activation does not require slowdown of the cell cycle and is regulated through the translation of maternally inherited mRNAs. Among these, the enhancer regulators P300 and Brd4 can prematurely activate transcription and restore transcriptional competency when maternal mRNA translation is blocked, whereas inhibition of histone acetylation blocks genome activation. We conclude that P300 and Brd4 are sufficient to trigger genome-wide transcriptional competency by regulating histone acetylation on the first zygotic genes in zebrafish. This mechanism is critical for initiating zygotic development and developmental reprogramming., This research was supported by the Fonds de Recherche du Québec Santé (postdoctoral fellowship to J.-D.B.), the Surdna Foundation and the Yale Genetics Venture Fund (postdoctoral fellowship to L.M.), and the NIH (grants R01 HD074078, GM103789, GM102251, GM101108, GM081602, R35 GM122580, and 4DNucleome program to A.J.G.). M.A.M.-M. was supported by Programa de Movilidad en Areas de Investigación priorizadas por la Consejería de Igualdad, Salud y Políticas Sociales de la Junta de Andalucía and is currently supported by Ramon y Cajal program from Ministerio de Ciencia, Innovación y Universidades of the Spanish government (RYC-2017-23041). The Giraldez lab is supported by the Howard Hughes Medical Institute Faculty Scholar program and was supported by the Pew Scholars Program in the Biomedical Sciences, March of Dimes 1-FY12-230, the Yale Scholars Program, and Whitman fellowship funds provided by E.E. Just, Lucy B. Lemann, Evelyn and Melvin Spiegel, The Haldan Keffer Hartline and Edward F. MacNichol, Jr., of the Marine Biological Laboratory in Woods Hole, Massachusetts to A.J.G.

e21036 Background: Lung cancer with drug receptor protein kinase fusion is regarded as a large class of molecular event, which is called fusion-dependent lung cancer. Fusion-dependent lung cancer accounts for about 10%-15% of non-small cell lung cancer. Therefore, the identification and detection of fusion-dependent lung cancer patients are very important in China. Here we reported a total of 4111 patients with Chinese fusion-dependent lung cancer. Methods: A multicenter study in China was initiated from Dec. 2019, and lung patients have been enrolled as of Dec. 2020. A total of 4111 patients with fusion-dependent were analyzed based on medical records and detailed patient questionnaires. Results: Of this entire cohort, 4111 patients were identified with fusion-dependent, including ALK fusions (2190, 53.27%), ROS1 fusions (894, 21.75%), RET fusions (521, 12.67%), NTRK fusions (128, 3.11%), FGFR fusions (84, 2.04%), NRG1 fusions (41, 1.00%), ERBB2 fusions (55, 1.34%), BRAF fusions (44, 1.07%), MET fusions (34, 0.83%), EGFR fusions (74, 1.80%), Other fusions (46, 1.12%). In ALK fusions, there were 1856 cases (84.75%) of EML4-, including A20(94.64%) and non-A20(5.36%), and other partners STRN-, KIF5B-, HIP1-, KLCl-, successively; In ROS1 fusions, there were 461 cases (51.57%) of CD74-, and other partners EZR-, SDC4-, SLC34A2-, TPM3-, GOPC-, successively; In RET fusions, there were 329 cases (63.15%) of KIF5B-, and other partners CCDC6-, NCOA4-, ERC1-, successively; In NTRK fusions, there were 22 cases (17.19%) of TPM3-NTRK1, and other types ETV6-NTRK3, LMNA-NTRK1, AGTPBP1-NTRK2 successively; In FGFR fusions, there were 40 cases (47.61%) of FGFR3-TACC3, and other types FGFR2-TACC2, ATE1-FGFR2, BAG4-FGFR1, successively; In NRG1 fusions, there were 11 cases (26.83%) of CD74-, and other partners SDC4-, RBPMS-, successively; In MET fusions, there were 9 cases (26.47%) of CD74-, and other partners HLA-DRB1-, KIF5B-, successively; In ERBB2, BRAF and EGFR fusions, the partners were scattered. And 3643 patiens (88.68%) of the detection were used DNA-based NGS, and Others were used RNA-based NGS. Patients with ALK and ROS1 fusions were routinely treated with targeted therapy. Patients with RET, NTRK, FGFR and NRG1 fusions were actively enrolled in the clinical trial. Patients with ERBB2, BRAF, MET and EGFR fusions were only treated with end-line trial target therapy, and they were eager to be included in the corresponding clinical trials. Conclusions: The common fusions of ALK, ROS1, RET are routine standardized treatment is very important. Although fusions are rare, it can not be ignored. Morever, these patients are eager to receive active targeted therapy. DNA+RNA based NGS is not widely used in China, but it is of great value to detect fusions, especially rare fusions, which is the future development trend.

Let G be a finite group, E a normal subgroup of G and p a fixed prime. We say that E is p-hypercyclically embedded in G if every p-G-chief factor of E is cyclic. A subgroup H of G is said to satisfy Π-property in G if |G∕K:NG∕K((H∩L)K∕K)| is a π((H∩L)K∕K)-number for any chief factor L∕K in G; we say that H has Π*-property in G if H∩Oπ(H)(G) has Π-property in G. In this paper, we prove that E is p-hypercyclically embedded in G if and only if some classes of p-subgroups of E have Π*-property in G. Some recent results are extended.

[EN] We give some sufficient conditions for a normal p-subgroup P of a finite group G to have every G-chief factor below it cyclic. The S-permutability of some p-subgroups of O^p(G)plays an important role. Some known results can be reproved and some others appear as corollaries of our main theorems., A. Ballester-Bolinches and R. Esteban-Romero have been supported by the Grant MTM2014-54707-C3-1-P from the Ministerio de Economia y Competitividad, Spain, and FEDER, European Union. A. Ballester-Bolinches and Y. Li have been supported by a project from the National Natural Science Foundation of China (NSFC, No. 11271085) and a project of Natural Science Foundation of Guangdong Province (No. 2015A030313791). L. Miao thanks the China Scholarship Council and for its financial support and the Department of Mathematics of the University of Valencia for its hospitality.

Recently, epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) have revolutionized non-small cell lung cancer (NSCLC) treatment. However, resistance remains a major obstacle. Anexelekto (AXL) is a member of receptor tyrosine kinases (RTKs) and shares the same downstream signaling pathways with EGFR, such as PI3K/AKT and MAPK/ERK. AXL overexpression in resistant tumors has been implicated in many previous studies in vitro and in vivo. In this study, we further examined whether expression of AXL and its downstream targets increased in gefitinib-resistant PC9 cells (PC9GR). In addition, we hypothesize that knocking down AXL in PC9GR and overexpressing AXL in PC9 using genetic tools can restore and decrease the sensitivity to gefitinib, respectively. We found that silencing AXL could sensitize the resistance to gefitinib, and the downstream pathways were significantly inhibited. Interestingly, we also discovered that increased AXL expression did promote the resistance, and its downstream targets were activated accordingly. Then 69 NSCLC patients who harbored EGFR mutation were recruited to analyze the expression of AXL and the association between AXL expression and clinical characteristics. We found that 5 of the 69 patients were AXL positive (about 7%), and AXL was related to tumor differentiation and tumor size. In this study, we concluded that the molecular mechanisms of AXL mediated resistance involved in the increased activity of the PI3K/AKT and MAPK/ERK1/2 pathways, and AXL overexpression could promote resistance, but it can be weakened when AXL expression is silenced.

Acquired resistance remains a key challenge in epidermal growth factor receptor (EGFR)‐tyrosine kinase inhibitors (TKIs) therapy in lung adenocarcinoma (LUAD). Recent studies have shown that Notch signaling is associated with drug resistance. However, its role and possible mechanisms in EGFR-TKI resistance are not yet clear. In our study, we found that among four members of NOTCH1‐4, only NOTCH3 was upregulated in LUAD tissues and TKI-resistant cell line (HCC827GR6). Knockdown of NOTCH3 by siRNA significantly inhibited proliferative ability, and decreased colony and sphere formation in HCC827GR6 cells. Then miR-150 was identified as a posttranscriptional regulator of NOTCH3. Its expression was downregulated in LUAD tissues and negatively correlated with NOTCH3 mRNA. The cell proliferation and IC50 of gefitinib were decreased in HCC827GR6 cells transfected with miR-150 mimic, but was reversed when cotransfected with NOTCH3 overexpressed vector. Moreover, we also enrolled 20 patients with advanced LUAD who have taken TKIs as first-line therapy in this study. We found that collagen 1A1 (COL1A1) expression was increased significantly in LUAD tissues both at mRNA and protein levels, and positively correlated with NOTCH3 expression verified in our data and TCGA data. Univariate survival analysis showed that patients with high protein expression of NOTCH3 or COL1A1 were associated with shorter overall survival (OS). Taken together, these results suggest that miR-150/NOTCH3/COL1A1 axis contributed to EGFR‐TKI resistance in LUAD, which provide a potential therapeutic target for LUAD treatment.

Cytoplasmic male sterility (CMS) lines are widely used for hybrid production in Brassica napus. The Shaan2A CMS system is one of the most important in China and has been used for decades; however, the male sterility mechanism underlying Shaan2A CMS remains unknown. Here, we performed transcriptomic and proteomic analysis, combined with additional morphological observation, in the Shaan2A CMS. Sporogenous cells, endothecium, middle layer, and tapetum could not be clearly distinguished in Shaan2A anthers. Furthermore, Shaan2A anther chloroplasts contained fewer starch grains than those in Shaan2B (a near-isogenic line of Shaan2A), and the lamella structure of chloroplasts in Shaan2A anther wall cells was obviously aberrant. Transcriptomic analysis revealed differentially expressed genes (DEGs) mainly related to carbon metabolism, lipid and flavonoid metabolism, and the mitochondrial electron transport/ATP synthesis pathway. Proteomic results showed that differentially expressed proteins were mainly associated with carbohydrate metabolism, energy metabolism, and genetic information processing pathways. Importantly, nine gene ontology categories associated with anther and pollen development were enriched among down-regulated DEGs at the young bud (YB) stage, including microsporogenesis, sporopollenin biosynthetic process, and tapetal layer development. Additionally, 464 down-regulated transcription factor (TF) genes were identified at the YB stage, including some related to early anther differentiation such as SPOROCYTELESS (SPL, also named NOZZLE, NZZ), DYSFUNCTIONAL TAPETUM 1 (DYT1), MYB80 (formerly named MYB103), and ABORTED MICROSPORES (AMS). These results suggested that the sterility gene in the Shaan2A mitochondrion might suppress expression of these TF genes in the nucleus, affecting early anther development. Finally, we constructed an interaction network of candidate proteins based on integrative analysis. The present study provides new insights into the molecular mechanism of Shaan2A CMS in B. napus.

QTL mapping for fiber-related traits and elucidation of a stable and novel QTL affecting seed lignin content, cellulose content and seed oil content. Dissection of the genetic networks for fiber biosynthesis is important for improving the seed oil content and meal value of Brassica napus. In this study, the genetic basis of seed fiber biosynthesis in B. napus was investigated via quantitative trait locus (QTL) analysis using a doubled haploid population derived from 'KenC-8' crossed with 'N53-2.' Seed lignin content (LC), cellulose content (CC) and hemicellulose content (HC) were significantly negatively correlated with seed oil content (OC). Co-localization QTLs among LC, CC, HC and OC on A09 were found with contributions ranging from 9.87 to 48.50%. Seven co-localization QTLs involved in the fiber component and OC were further verified by bulked segregant analysis (BSA). The unique QTL uqA9-12 might be a real and new QTL that was commonly identified by QTL mapping and BSA and simultaneously affected LC, CC and OC with opposite additive effects. A potential regulatory network controlling seed fiber biosynthesis was constructed to dissect the complex mechanism of seed fiber and oil accumulation, and numerous candidate genes were identified in the fiber-related QTL regions. These results provided an enrichment of QTLs and potential candidates for fiber biosynthesis, as well as useful new information for understanding the complex genetic mechanism underlying rapeseed seed fiber accumulation.

Background Rapeseed (Brassica napus) is an important oil seed crop in the Brassicaceae family. Chemical induced male sterility (CIMS) is one of the widely used method to produce the hybrids in B. napus. Identification of the key genes and pathways that involved in CIMS were important to understand the underlying molecular mechanism. In the present report, a multi-omics integrative analysis, including of the proteomic, transcriptomic and miRNAs, combined with morphological and physiological analysis were conducted. Results Earlier degeneration of the tapetosomes and elaioplasts, aberrantly stacking in tapetal cells and incompletely deposition in tryphine of pollen wall were observed in chemical hybridization agent (CHA) of SX-1 treated B. napus through SEM and TEM analysis. It was revealed that the deficiencies in protein processing in endoplasmic reticulum (ER) and flavonoids biosynthesis were occurred at early stage in the SX-1 treated materials. Subsequently, plant hormone signal transduction, biosynthesis of amino acids, fatty acids and steroid in anther at later stages were identified down-regulated after SX-1 treatment. 144 transcript factors (TFs) were also indentified to down-regulated at early stage, which suggested the early regulation in anther and pollen wall development were disordered in CHA treated B. napus. In addition, 7 important miRNAs were identified and 2 of the predicted target genes of miRNAs were Rf-like genes. Conclusions Taken together, an interaction network of candidate genes and the putative metabolism pathways were constructed based on the multi-omics integrative analysis, it provided a new insight into the male sterility induced by CHA of SX-1 in B. napus. Electronic supplementary material The online version of this article (10.1186/s12864-018-5203-y) contains supplementary material, which is available to authorized users.

The awakening of the zygote genome, signaling the transition from maternal transcriptional control to zygotic control, is a watershed in embryonic development, but the factors and mechanisms controlling this transition are still poorly understood. By combining CRISPR-Cas9-mediated live imaging of the first transcribed genes (miR-430), chromatin and transcription analysis during zebrafish embryogenesis, we observed that genome activation is gradual and stochastic, and the active state is inherited in daughter cells. We discovered that genome activation is regulated through both translation of maternal mRNAs and the effects of these factors on the chromatin. We show that chemical inhibition of H3K27Ac writer (P300) and reader (Brd4) block genome activation, while induction of a histone acetylation prematurely activates transcription, and restore genome activation in embryos where translation of maternal mRNAs is impaired, demonstrating that they are limiting factors for the activation of the genome. In contrast to current models, we do not observe triggering of genome activation by a reduction of the nuclear-cytoplasmic (N/C) ratio or slower cell division. We conclude that genome activation is controlled by a time-dependent mechanism involving the translation of maternal mRNAs and the regulation of histone acetylation through P300 and Brd4. This mechanism is critical to initiating zygotic development and developmental reprogramming.

Background Chemotherapy-related hepatic dysfunction affected the prognosis of non-small cell lung cancer (NSCLC). However, predictive factors of chemotherapy-related hepatic dysfunction remained undefined. Objective To identify the predictive factors for hepatic dysfunction during cytotoxic chemotherapy in Chinese patients with advanced NSCLC. Methods We retrospectively reviewed the medical records of patients with advanced NSCLC who received cytotoxic chemotherapy at Division of Respiratory Medicine, the affiliated Drum Tower Hospital of Nanjing University, from July 2012 to January 2015. We investigated the incidence of hepatic dysfunction during chemotherapy and evaluated several clinical factors that are associated with hepatic dysfunction, including body mass index (BMI) and high density lipoprotein cholesterol (HDL-C). Results A total of 116 patients were enrolled in study, 54 patients (46.57%) experienced hepatic dysfunction after receiving chemotherapy. Multivariate analysis for hepatic dysfunction in patients with advanced NSCLC showed that hepatic dysfunction was associated with higher BMI (odds ratio = 4.742, P = 0.001) and lower HDL-C (odds ratio = 3.018, P = 0.019). Pearson's rank correlation analysis revealed that HDL-C and BMI presented a negative correlation in patients with hepatic dysfunction (r = -0.487, P Conclusions Higher BMI and lower HDL-C levels seem to be good independent predictive factors for chemotherapy-related hepatic dysfunction in advanced NSCLC. In addition, a negative correlation was presented between BMI and HDL-C.

To the Editor: Pulmonary Langerhans cell histiocytosis (PLCH) in adults is a rare disease occurring almost exclusively in smokers. The characteristic high-resolution computed tomography (HRCT) manifestation of PLCH is a combination of cysts (or cavities) and nodules mainly in the upper lung zone.[1] However, not all HRCT patterns of PLCH are typical. Few treatments are effective in current practice regarding PLCH. Targeted therapy with an inhibitor of mutated BRAF (vemurafenib) has been proved effective in Langerhans cell histiocytosis (LCH) harboring BRAF valine at position 600 (V600E) mutation.[2] MAP2K1 mutations are mutually exclusive with BRAF mutations and might have implications for the use of BRAF targeted therapy.[3] Here, we reported a case of PLCH proven by lung biopsy. A 61-year-old man was referred to our institution for evaluation of intermittent cough with white phlegm. He had been a former smoker (cumulative dose = 20 pack/years) and quit approximately one year before. A chest HRCT scan [Figure 1a] revealed, besides emphysema, diffuse nodules up to 1.5 cm in diameter spread in both lung fields without associated lymphadenopathy or pleural effusions. Positron emission tomography-computed tomography (CT) showed diffuse nodules in lung, many of which revealed increased 18F-deoxyglucose uptake (the average standardized uptake value [SUV]: 2.3; maximum SUV: 3.0). Pulmonary function test revealed moderate mixed restriction and obstruction pattern. Routine laboratory tests, ultrasound cardiogram, and bronchoscopy were unremarkable. CT-guided percutaneous transthoracic fine-needle aspiration (FNA) of pulmonary lesions was performed twice, showing atypical epithelial cells. The patient was discharged upon his own request. Figure 1 (a) Chest high-resolution computed tomography findings of the patient. (b) Histopathological findings of the lung biopsy specimen. H and E stain showed cellular nodule with aggregates of Langerhans cells (original magnification ×200); immunohistochemistry ... Two months later, reexamined chest HRCT [Figure 1a] showed a marked increase in the number and size of nodules, with diameter up to 2.0 cm. The patient underwent video-assisted thoracoscopic surgical (VATS) biopsy of the left upper and lower lobes. During the procedure, many palpable nodules were noted in lung parenchyma with diameters ranging from 0.3 to 1.8 cm. A key morphological feature of LCH cells on microscopic examination was noted, which was their highly convoluted nuclear membranes, and the cells strongly expressed Cluster of Differentiation (CD)1a and S100 [Figure 1b]. PLCH diagnosis was made. Treatment started with prednisone (PDN) (0.5 mg·kg−1·d−1 for 1 month). Unfortunately, chest HRCT obtained one month later showed an increase in the number and size of nodular lesions [Figure 1a]. Based on the results of LCH-III,[4] a randomized international clinical trial embarked by the Histiocyte Society, an adjusted chemotherapeutic regimen of methotrexate (MTX) + vindesine (VDS) + PDN was pursued, i.e., MTX 500 mg/m2 once every two weeks intravenous (i.v.), VDS 3 mg/m2 once a week i.v., PDN 30 mg/m2 d1-30 orally (afterward weekly reduction). He experienced WHO Grade 1/2 toxicities including hepatotoxicity, nausea, and vomiting. One month later, chest HRCT showed a partial clearing of nodules [Figure 1a], and his cough resolved. To explore whether the patient would benefit from the BRAF inhibitor therapy, BRAF and MAP2K1 mutations were further tested. Deoxyribonucleic acid (DNA) was extracted from formalin-fixed and paraffin-embedded (FFPE) samples of the surgical biopsy after histological detection of histiocyte-rich areas, using QIAmp DNA FFPE tissue kit (QIAGEN, Germany) according to instructions. Exon 2 and exon 3 of MAP2K1 and exon 15 of BRAF were amplified by polymerase chain reaction and sequenced with primers as listed in Table 1. As indicated in Figure 1c, one silent mutation (code “CAG” to “CAA” at position 610) in exon 2 of MAP2K1 was detected. However, there were no mutations in exon 3 of MAP2K1 and exon 15 of BRAF. The experiment was approved by the Ethics Committee of Drum Tower Hospital, and the patient gave written informed consent. Table 1 Polymerase chain reaction and sequencing primers for MAP2K1 (exon 2 and exon 3) and BRAF (exon 15) Several findings of this case were quite unique and rare. First, the chest HRCT manifestation of this patient was diffuse nodules alone throughout the lung, without formation of cysts or cavities. Second, the onset of disease was nearly one year after smoking cessation. Diffuse lung nodules progressed both in number and size despite smoking cessation. Third, corticosteroid monotherapy failed to control disease progression while intensified combination chemotherapy (MTX + VDS + PDN) showed efficacy. Finally, and particularly, BRAF and MAP2K1 of the lung lesions were sequenced. One MAP2K1 mutation was observed while BRAF V600E mutation was not detected. To the best of our knowledge, the report regarding MAP2K1 mutations in single-system PLCH was seldom and a new mutation site was identified (code “CAG” to “CAA” at position 610 in exon 2 of MAP2K1). These findings differentiate this case from the usual case of PLCH seen in clinical practice. PLCH usually has little systemic damage. In this case, we did not find extrapulmonary disease manifestations. In elder patients with pure nodular forms in chest HRCT, it is especially important to exclude malignancy, which is why this patient received twice CT-guided percutaneous transthoracic FNA followed by VATS lung biopsy. It is generally thought that lung bases are spared in PLCH. However, in this patient, nodules extended into the lower lung zones and costophrenic angles were infiltrated, which might associate with the severity of disease. Persisting uncertainty about the pathogenesis of PLCH has limited current treatment alternatives. Smoking cessation has been regarded as a treatment; however, worsening conditions despite cessation was observed in this case, indicating that PLCH might be induced by antigens in cigarette smoke but not terminated by smoke cessation. The role of smoking cessation on disease progress seems indifferent. In this case, corticosteroid monotherapy failed to control the disease. This patient further received chemotherapy (MTX + VDS + PDN) according to LCH-III.[4] HRCT imaging showed improvement one month later. Even though this case is one-system LCH, involving the lung might associate with rapid progression and worse prognosis, thus a more intensive chemotherapeutic regimen might be necessary as in multisystem LCH. The most common activating somatic mutation in BRAF gene is a substitution of glutamic acid for V600E, which has been identified in a part of LCH cases. This mutation results in constitutive activation of the mitogen-activated protein kinase (MAPK) pathway. MAP2K1 encodes the dual-specificity kinase of the MAPK pathway. MAP2K1 mutations have been demonstrated to confer resistance to BRAF inhibitor therapy in LCH and other neoplasms.[5] A mutation in exon 2 of MAP2K1 was positive in this patient. Even though it is a silent mutation, it indicates there might be another mechanism of MAPK pathway activation in BRAF V600E-negative PLCH. There is clearly a necessity for better therapies for PLCH. Financial support and sponsorship This work was supported by the grants from The Natural Science Foundation of Jiangsu Province (No. BK20130089), and the National Natural Science Foundation of China (No. 81501972). Conflicts of interest There are no conflicts of interest.

Background: Acute fibrinous and organizing pneumonia (AFOP) is a unique pathological entity with intra-alveolar fibrin in the form of fibrin balls and organizing pneumonia. It was divided into rare idiopathic interstitial pneumonia according to the classification notified by American Thoracic Society/European Respiratory Society in 2013. As a rare pathological entity, it is still not well known and recognized by clinicians. We reviewed the clinical features of 20 patients with AFOP diagnosed in a teaching hospital. Methods: The medical records of 20 patients with biopsy-proven diagnosis of AFOP were retrospectively reviewed. The patients' symptoms, duration of the disease, comorbidities, clinical laboratory data, pulmonary function testing, radiographic studies, and the response to treatment were extracted and analyzed. Results: Fever was the most common symptom and was manifested in 90% of AFOP patients. For clinical laboratory findings, systematic inflammatory indicators, including C-reactive protein and erythrocyte sedimentation rate, were significantly higher than normal in AFOP patients. In accordance with this increased indicators, injured liver functions were common in AFOP patients. Inversely, AFOP patients had worse clinical conditions including anemia and hypoalbuminemia. For pulmonary function testing, AFOP patients showed the pattern of restrictive mixed with obstructive ventilation dysfunction. For high-resolution computerized tomography (HRCT) findings, the most common pattern for AFOP patients was lobar consolidation which was very similar to pneumonia. However, unlike pneumonia, AFOP patients responded well to glucocorticoids. Conclusion: Patients with AFOP manifest as acute inflammatory-like clinical laboratory parameters and lobar consolidation on HRCT, but respond well to steroid.

Overcoming EGFR-TKI resistant which has the initial enthusiasm over substantial clinical responses is a formidable challenge on nowadays. In this study, we showed that cholesterol level in lipid rafts in gefitinib resistant non-small cell lung cancer (NSCLC) cell lines was remarkably higher than gefitinib sensitive cell line, and depletion of cholesterol increased gefitinib sensitivity. Furthermore, cholesterol-depleted enhanced gefitinib inhibit phosphorylation of EGFR, Akt-1, MEK1/2, and ERK1/2 and these were reversed in cholesterol add-back experiments. Gefitinib resistant cell lines showed high affinity of gefitinib and EGFR when cholesterol was depleted. Therefore, targeting cholesterol combined with EGFR-TKI is potentially a novel therapeutic strategy for gefitinib resistant treatment.

e14739 Background: CZ was recommended by NCCN for ALK-rearranged metastasis NSCLC as first-line therapy. Knowledge on the occurrence of ALK actionable muts within cancer types and the responses of ALK fusion types to CZ could better guide the treatment of ALK-rearranged pts. Methods: 772 ALK-mutated pts (with any ALK muts) detected by tissue/ctDNA NGS in our institution from Oct. 2016 to Oct. 2018 were included. ALK+ (with any ALK actionable muts) and EGFR- (with no EGFR actionable muts) pts treated with CZ at any line were included for survival analysis. Progress-free survival (PFS) was estimated using Kaplan-Meier method and compared using log-rank test. Results: 339 ALK+ pts were analyzed, including 331 lung cancer (LC: 265 NSCLC; 1 SCLC; 65 Unknown histology), 3 brain cancer (BC), 1 colon cancer (CC), 3 cancer of unknown primary (CUP), and 1 with no clinical information. The most common ALK mut was fusion (331/339), including 315 EML4-ALK, 5 KIF5B-ALK, 4 STRN-ALK, 1 ACSL3-ALK, 1 CLIP4-ALK, 1 DCTN1-ALK, 1 HIP1-ALK, 1 SRBD1-ALK, 1 TFG-ALK, and 1 ZFP161-ALK. Fusion position was known for 308 EML4-ALK, including 132 V1, 108 V3, 32 V2, 19 V5’, and 17 other variants (Vx). Except for 2 V1 with CUP, 1 KIF5B-ALK with CUP, 1 Vx with CC, and 1 EML4-ALK (no variant information) with BC, all the other fusions occurred in LC. ALK F1174L was detected only in 2 pts, both of whom had BC, specifically neuroblastoma. CZ was given to 76 ALK+ EGFR- pts (mPFS: 9 mo, 95% CI: 8 – 13), including 70 EML4-ALK and 6 non- EML4-ALK (mPFS: 10 vs. 9 mo, p = 0.7). Though non-significant, V1 (n = 32) on average had a longer mPFS than V3 (n = 28) (12 vs. 8 mo, p = 0.3), which may partially be explained by the younger age at diagnosis of V1 than V3 (48 vs. 53, p = 0.005). 1 TFG-ALK, 1 CLIP4-ALK, 1 DCTN1-ALK, 1 HIPI-ALK, 1 ZFP161-ALK, and 1 KIF5B-ALK achieved a PFS of 3, 4, 8, 9, 13, and 19 mo on CZ, respectively. 1 SCLC (V3) received CZ at first-line progressed after 6 mo. Conclusions: Though majority of the ALK actionable muts were commonly found in LC, some alterations may occur more often in other cancer types, such as F1174L in neuroblastoma. Some fusion types (e.g. TFG-ALK) may benefit less from CZ than others (e.g. KIF5B-ALK). Limited by relative small sample size, these results merit further validation and investigation.

e14543 Background: Somatic mutations in MET are diverse and include MET amplification, exon 14 skipping and also some functional point mutations (H1094Y, Y1230H, D1228N, etc). Generally, amplification and ex14 skipping are the two main mutation types. Methods: MET mutation profiles were analyzed using hybridization capture based next-generation sequencing (NGS), which enables the simultaneous detection of multi-mutation types of at least 59 genes (range 59-1021 genes). Results: Screening tens of thousands treatment naive patients, we found 319 patients with MET mutations, 56 patients (18%) with MET amplification, 55 patients (17%) with MET ex14 skipping, and 214 patients with other unknown functional point mutations. 39 cases with MET amplification and 52 cases with MET ex14 skipping in lung cancer. Patients with MET amplification or ex14 skipping had fewer mutations compared with patients with other MET mutations ( P < 0.0001), while there were no differences in ctDNA abundance between two groups ( P = 0.58). Further compared the MET amplification and ex14 skipping groups, we found mutation numbers and ctDNA abundance were both significant lower in ex14 skipping group (mutation numbers: P = 0.0134; ctDNA abundance: P < 0.0001). MET amplification range from 2 to 7 copies in these 39 patients, and patients with more than 3 copies had a higher ctDNA abundance than = < 3 copies patients ( P < 0.0005). Six patients with both amplification and ex 14 skipping, all of them with advanced age ( > 70 years). Of the MET amplification group, 11 patients (28%) developed EGFR driver mutation concurrently, four patients with L858R, three with ex19 del, one with G719A, one with both L858R and EGFR amplification, one with both ex19 del and amplification and one with only amplification, while no patients were found to have any EGFR driver mutation in MET ex14 skipping group. Conclusions: The cfDNA from patients with EGFR amplification or ex14 skipping was only 18% or 17% in MET mutations. Both of these two mutation types were mainly found in lung cancer. Furthermore, MET amplification group patients had fewer mutations and lower ctDNA abundance. In treatment naïve patients, MET amplification concurrent with other EGFR drive mutations, rather than ex14 skipping.

Although the correlation between glypican-5 (GPC5) and lung cancer is well known, the effect of GPC5 expression on non-small cell lung cancer (NSCLC) survival remains to be determined. In the present study, GPC5 expression in A549, H3255, and SPC-A1 NSCLC cell lines was evaluated by reverse transcription-polymerase chain reaction (RT-PCR) and western blot analysis. GPC5 mRNA and protein expression levels were found to be higher in A549 and H3255 cells compared with SPC-A1 cells. The role of GPC5 in NSCLC cell migration was evaluated in vitro by shRNA-mediated knockdown or the overexpression of GPC5 through scratch and transwell assays. The mean migration rates of cancer cells transfected with pRNAT-shRNA-GPC5-1 were reduced compared with the controls in A549 (P

Lung cancer is the leading cause of cancer-related death worldwide. Overall survival of all stages is unsatisfactory due to high metastasis and recurrence rates. Recent studies indicated that epithelial-to-mesenchymal transition (EMT) is involved in the progression and metastasis in cancer. Some reports also indicate that HS3ST3B1 is involved in angiogenesis and the proliferation of cancer cells. However, its role in non-small cell lung cancer (NSCLC) is not well known. In this study, we found that HS3ST3B1 was significantly up-regulated in NSCLC tissues compared with matched normal tissues (P=0.02). Its expression was also up-regulated in mesenchymal phenotype of NSCLC cell lines compared with epithelial phenotype (P

An increasing number of deregulated long non-coding RNAs (lncRNA) have been implicated in cancer in humans, suggesting that lncRNAs may be involved in tumorigenesis or tumor progression. In previous investigations, lncRNA, AFAP1-AS1 has been found to be associated with several cancers, including nasopharyngeal carcinoma, lung cancer and esophageal adenocarcinoma. However, the function of AFAP1-AS1 in lung cancer has not been reported. In our present study, we found that AFAP1-AS1 was overexpressed in lung adenocarcinoma and associated with survival time. The low expression of AFAP1-AS1 was an independent predictor for disease-free survival in patients with lung adenocarcinoma. Additionally, we transfected AFAP1-AS1 siRNA into H1975 and HCC827, both lung adenocarcinoma cancer cells, and found that cell growth was suppressed, apoptosis induced and invasion inhibited. Taken together, AFAP1-AS1 down-regulation exerts growth suppression and apoptosis induction in lung adenocarcinoma cells, suggested it may participate in turmorigenesis and be a therapeutic target for treating lung adenocarcinoma in future.

12101Background: Tumor mutation burden (TMB) is new clinical marker that predicts responses to immunotherapy, but the molecular basis of TMB is unknown. Understand the association between genomic l...

9068Background: AZD9291, a third-generation epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI), is active against patients with EGFR T790M-mutant non–small cell lung cancer (NS...

e24267Background: Approximately 4-7% of non-small cell lung cancer (NSCLC) patients have abnormal activation of MET, and MET inhibitors have shown promising results. However, the prevalence of MET ...

Yang Lv,1,2 Li-Yun Miao,2 Qiu-Fang Chen,1 Yan Li,2 Zhi-Xiang Shi,1 Xuan-Sheng Ding1 1Department of Clinical Pharmacy, China Pharmaceutical University, Nanjing, Jiangsu, People’s Republic of China; 2Division of Respiratory Medicine, Department of Respiration, The Affiliated Drum Tower Hospital of Nanjing University Medical College, Nanjing University Medical School, Nanjing, Jiangsu, People’s Republic of China Abstract: High-density lipoprotein cholesterol (HDL-C) has an inverse association with the incidence of lung cancer. However, whether it can be used as a predictive factor in advanced lung adenocarcinoma patients treated with epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKI) still remains undefined. This research aimed at studying the relationship of serum HDL-C baseline level and HDL-C kinetics to EGFR mutation, the efficacy of EGFR-TKI, and the predictive value of PFS. The presence of mutation rate in the 192 patients with lung adenocarcinoma was compared within stratified groups. Levels of baseline HDL-C and kinetics of HDL-C were analyzed retrospectively in patients treated with EGFR-TKI harboring EGFR mutation. Univariate and multivariate analyses were performed to investigate the prognostic value of HDL-C. EGFR mutation rate of HDL-C high-level group was significantly higher than that of low-level group (59.0% vs 35.6%, P=0.001). Multivariate logistic analysis showed that high-level HDL-C was an independent predictive factor for EGFR gene mutation (P=0.005; odds ratio =0.417; 95% confidence interval [CI], 0.227–0.768). Patients with a low level of HDL-C before therapy showed a progression of disease in most cases (P

To investigate the clinical and high-resolution computed tomography (HRCT) features of the welder's pneumoconiosis.A total of 10 patients diagnosed by bronchoscopy and pathology from January 2010 to January 2015 in Nanjing Drum Tower Hospital were recruited in this study, and the clinical manifestations, pulmonary function tests, pathology and HRCT data were collected and analyzed retrospectively.Ten patients all had welder's occupational history for 5-30 years, with the main clinical manifestations of cough, sputum production, chest tightness and other symptoms. And the ratio of forced expiratory volume in one second and forced vital capacity (FEV1/FVC) of 4 patients was lower than 75.0% in the pulmonary function tests. Transbronchial lung biopsy specimen showed numerous hemosiderin-laden macrophages within the alveolar space, associated with positive iron staining. In welder's pneumoconiosis, small centrilobular nodules (10 cases) were frequently seen on HRCT in bilateral lung fields, with branching linear structures or the tree buds like shadows in 7 cases; 3 patients also showed areas of ground-glass attenuation. And the patients were misdiagnosed as tuberculosis (1/10), interstitial pneumonia (3/10), allergic alveolitis (1/10), diffuse lung disease (2/10), and vasculitis (1/10).Clinical manifestations of welder's pneumoconiosis are not specific; small centrilobular nodules are frequently seen on HRCT, with linear opacities or tree bud-like shadows.

Fully grown oocytes of most vertebrates are arrested at prophase I of meiosis (G2 arrest). Upon exposure to steroid hormones, oocytes resume meiotic process, also called G2/M transition. The G protein-signaling pathway has been shown to play essential roles in the meiotic arrest at G2 phase. Previously, we showed that long chain fatty acyl-coenzyme A synthetase acsl1b was required for maintaining the meiotic arrest in Xenopus Acsl1b presumably synthesizes palmitoyl-coenzyme A that can be utilized by acyltransferases to modify proteins essential for the G2 arrest. In the present study, we report that protein acyltransferase ZDHHC3 functions downstream of acsl1b to maintain oocyte meiotic arrest. Depletion of maternal ZDHHC3 RNA in oocytes reduces the progesterone threshold to promote G2/M transition from 2 to 0.01 μM. As expected, Gs alpha palmitoylation level is greatly decreased in ZDHHC3-depleted oocytes. Furthermore, we mapped ZDHHC3 palmitoylation sites in Gs alpha and showed that palmitoylat...

Large numbers of maternal RNAs are deposited in oocytes and are reserved for later development. Control of maternal RNA translation during oocyte maturation has been extensively investigated and its regulatory mechanisms are well documented. However, translational regulation of maternal RNAs in early oogenesis is largely unexplored. In this study, we generated zebrafish zar1 mutants which result in early oocyte apoptosis and fully penetrant male development. Loss of p53 suppresses the apoptosis in zar1 mutants and restores oocyte development. zar1 immature ovaries show upregulation of proteins implicated in endoplasmic reticulum (ER) stress and the unfolded protein response (UPR). More importantly, loss of Zar1 causes markedly upregulation of zona pellucida (ZP) family proteins, while overexpression of ZP proteins in oocytes causes upregulation of stress related activating transcription factor 3 (atf3), arguing that tightly controlled translation of ZP proteins is essential for ER homeostasis during early oogenesis. Furthermore, Zar1 binds to zona pellucida (zp) mRNAs and represses their translation. Together our results indicate that regulation of translational repression and de-repression are essential for precisely controlling protein expression during early oogenesis.

To investigate the difference in clinical features and radiologic findings between patients with cryptogenic organizing pneumonia (COP) and connective tissue disorder related organizing pneumonia (CTD-OP).A total of 30 subjects with COP and 22 subjects with CTD-OP collected from 2005 to 2013 were retrospectively reviewed in the Affiliated Nanjing Drum Tower Hospital of Nanjing University Medical School, and the diagnosis of all patients were confirmed by lung biopsy.The common underlying diseases in patients with CTD-OP were Sjogren syndrome(SS), poly-/dermatomyositis(PM/DM), rheumatoid arthritis (RA). There were no significant differences in clinical manifestations between CTD-OP and COP. Compared with COP patients, the proportion of female patients was higher in CTD-OP. Higher positive rate for ANA was found in CTD-OP group (CTD-OP:63.6%; COP:10.0%; P0.01). There were no significant differences in parameters of lung function between CTD-OP and COP. As to radiological findings, the most common patterns were multiple patchy, linear shadows and ground-glass opacity. Some patients showed solitary nodule or consolidation and pleural effusion. Reticular shadow was a rare pattern among these patients. Most lesions were under the pleura and/or around the bronchus.There are no significant differences in clinical and radiologic manifestations between COP and CTD-OP, except that the proportion of women and ANA positive rate were higher in CTD-OP.

Although associations between thioredoxin interacting protein (TXNIP) and cancers have been recognized, the effects of TXNIP on non-small cell lung cancer (NSCLC) prognosis remained to be determined in detail. In addition, while hypoxia is a key characteristic of tumor cell growth microenvironment, the effect of hypoxia on TXNIP expression is controversial. In this study, formaldehyde fixed and paraffin embedded (FFPE) samples of 70 NSCLC patients who underwent resection between January 2010 and December 2011 were obtained. Evaluation of TXNIP and hypoxia inducible factor-1α (HIF-1α) protein expression in FFPE samples was made by immunohistochemistry. By Kaplan-Meier method, patients with high TXNIP expression demonstrated a significantly shorter progression free survival (PFS) compared with those with low TXNIP expression (18.0 months, 95%CI: 11.7, 24.3 versus 23.0 months, 95%CI: 17.6, 28.4, P=0.02). High TXNIP expression level was also identified as an independent prognostic factor by Cox regression analysis (adjusted hazard ratio: 2.46; 95%CI: 1.08, 5.56; P=0.03). Furthermore, TXNIP expression was found to be significantly correlated with HIF- 1α expression (Spearman correlation=0.67, P=0.000). To further confirm correlations, we established a tumor cell hypoxic culture model. Expression of TXNIP was up-regulated in all three NSCLC cell lines (A549, SPC-A1, and H1299) under hypoxic conditions. This study suggests that hypoxia induces increased TXNIP expression in NSCLC and high TXNIP expression could be a poor prognostic marker.