Lung adenocarcinoma (LUAD) is a highly malignant tumor with a dismal prognosis worldwide. Glial maturation factor β (GMFB), initially identified as a regulator of glial cell development, has recently been implicated in tumor microenvironment-mediated carcinogenesis. This study aims to validate the role of GMFB in LUAD progression and explore its potential as a novel biomarker and therapeutic target. GMFB expression was evaluated in LUAD tissues and cell lines using bioinformatics analysis, immunohistochemistry (IHC), Western blotting (WB), and quantitative real-time PCR (qRT-PCR). In vitro functional assays (CCK-8, EdU, colony formation, transwell, wound healing) and a nude mouse subcutaneous tumor model were used to assess the effects of GMFB on cell proliferation and migration. RNA sequencing and bioinformatics were employed to identify downstream pathways, with rescue experiments confirming mechanistic insights. GMFB was significantly overexpressed in LUAD tissues and cell lines, and high GMFB expression was associated with poor patient survival. Functional studies showed that GMFB knockdown attenuated LUAD cell proliferation and migration, whereas GMFB overexpression promoted these phenotypes. In vivo experiments confirmed GMFB-driven tumor growth. Mechanistically, RNA sequencing and WB analysis revealed that GMFB activated the MAPK/ERK signaling pathway, and pharmacological inhibition of ERK reversed GMFB-mediated tumor progression. GMFB promotes LUAD progression by activating the MAPK/ERK pathway, positioning it as a potential biomarker and therapeutic target for LUAD diagnosis and treatment.

Lung adenocarcinoma(LUAD) is the primary reason for cancer-related deaths globally. GINS1 has a significant regulatory function in DNA replication. It is overexpressed in various malignant tumors, but the specific molecular mechanisms of GINS1 in LUAD pathogenesis are not fully elucidated. This is the first report that GINS1 enhances LUAD by activating Wnt/β-catenin signaling pathway, and may serve as a potential target for therapy. Bioinformatic analysis including analysis of difference, survival analysis and pathway enrichment, immunohistochemistry(IHC), western blotting(WB), and quantitative real time polymerase chain reaction(qRT-PCR) were used to detect GINS1 expression in LUAD cell lines and tissues. A range of in vivo and in vitro experiments, such as cck-8, EdU, cloning experiment, wound healing experiment and transwell experiment, confirmed that GINS1 facilitated the proliferation and migration of LUAD. Additionally, the potential mechanism of GINS1 was hypothesized through WB and transcriptome sequencing. The rescue experiment was used to verify our conclusion. In this study, we discovered that GINS1 is significantly overexpressed in LUAD cell lines and tissues. Analysis of Kaplan − Meier survival data indicated that high levels of GINS1 expression are often linked to unfavorable survival outcomes. Additionally, a series of experiments showed that silencing GINS1 led to less proliferation and migration of LUAD cell lines, while its overexpression enhanced tumor progression. Furthermore, subcutaneous tumor experiments in nude mice supported the role of GINS1 in promoting tumor development in vivo. Lastly, transcriptome sequencing revealed that tumor progression is related to cell cycle (G1 to S phase transition associated with cyclinD) and β-catenin signaling pathway, which we subsequently validated using WB. A series of rescue experiment further confirmed that GINS1 facilitates the advancement of LUAD via the β-catenin signaling pathway. Our findings suggest that GINS1 plays a critical role in the progression of LUAD by modulating key molecular pathways, particularly the β-catenin signaling pathway., and it might serve as a potential new target of β-catenin signaling pathway for treatment of LUAD.

Background Children with acute lymphoblastic leukemia (ALL) are highly susceptible to infections due to active leukemia and chemotherapy-induced immunosuppression during the induction phase. Knowledge on the incidence, impact and outcome of infection in children during the induction phase of treatment without prophylactic antibiotics and antifungals is currently limited. Methods The clinical data of pediatric patients with newly diagnosed ALL at Qilu Hospital of Shandong University between January 1, 2015 and December 31, 2019 were reviewed and analyzed. Results Of the 377 children in the retrospective cohort, 237 developed 259 episodes of infections in different sites. The most common infections were respiratory tract infections followed by fever of unknown origin. A total of 65 microbial strains were detected, including 39 strains of Gram-positive (G+) bacteria, 17 strains of Gram-negative (G−) bacteria, five strains of fungi and four strains of viruses. Staphylococcus was the predominant genera of G+ bacteria, whereas Pseudomonas aeruginosa was the common genera of G− bacteria. There were significant differences between infection and time from symptom onset to diagnosis (≤ 15d or > 15d), duration of granular deficiency (≥ 15d or < 15d), and whether protective isolation was given in laminar beds (P < 0.05). Of the 237 episodes of infections, 45 patients had delayed CAM chemotherapy and seven patients died. Conclusion Abnormal bone marrow hematopoiesis and high incidences of infection, hemorrhage, and anemia were present in pediatric patients during the induction phase. In addition, infection-related morbidity, mortality, and chemotherapy delay remain clinically significant.

In this work, ginsenoside CK was confirmed as an EGFR tyrosine kinase inhibitor, with an IC50 value of 32.58 +/- 0.09 mu M. Molecular docking showed that ginsenoside CK could fit into EGFR binding pocket, thereby acting as a ligand for EGFR. Ginsenoside CK inhibited proliferation of A549 cells as well as induced its apoptosis via downregulating anti-apoptotic factors and upregulating apoptosis induction factors. Ginsenoside CK alone and combined with gefitinib inhibited the tumor progression of A549 lung cancer xenografts in BALB/c nude mice. ELISA assay showed that ginsenoside CK combined with gefitinib alleviated the inflammatory response by suppressing the release of pro-inflammatory cytokines, indicating better inhibitory effects than ginsenoside CK or gefitinib alone. This work further confirmed that ginsenoside CK regulated the EGFR/MAPK/ERK signaling both in vitro and in vivo. In conclusion, ginsenoside CK can exhibit the anti-lung cancer effect via acting as an EGFR tyrosine kinase inhibitor.

Acute myeloid leukemia (AML) is an aggressive hematological tumor caused by the malignant transformation of myeloid progenitor cells. Although intensive chemotherapy leads to an initial therapeutic response, relapse due to drug resistance remains a significant challenge. In recent years, accumulating evidence has suggested that post-transcriptional methylation modifications are strongly associated with tumorigenesis. However, the mRNA profile of m7G modification in AML and its role in drug-resistant AML are unknown. In this study, we used MeRIP-seq technology to establish the first transcriptome-wide m7G methylome profile for AML and drug-resistant AML cells, and differences in m7G between the two groups were analyzed. In addition, bioinformatics analysis was conducted to explore the function of m7G-specific methylated transcripts. We found significant differences in m7G mRNA modification between AML and drug-resistant AML cells. Furthermore, bioinformatics analysis revealed that differential m7G-modified mRNAs were associated with a wide range of cellular functions. Importantly, down-methylated m7G modification was significantly enriched in ABC transporter-related mRNAs, which are widely recognized to play a key role in multidrug resistance. Our results provide new insights into a novel function of m7G methylation in drug resistance progression of AML.

e20513 Background: Lung cancer is the most incident worldwide, and surveillance of recurrence remains a clinically unmet need. Non-invasive early detection is essential to improve prognosis of lung cancer. DNA methylation-based biomarkers for early cancer detection is promising. This study aims to investigate the methylation biomarkers of ctDNA that could predict the postoperative recurrence in early-stage lung cancer. Methods: The HM450K DNA methylation microarray data of lung adenocarcinoma (LUAD) (492 tumor tissues and 32 normal tissues) and lung squamous cell carcinoma (LUSC) (415 tumor tissues and 43 normal tissues) were downloaded from TCGA Data Portal ( https://portal.gdc.cancer.gov/ ), and healthy individuals were obtained from GEO database (656 whole blood from GSE40279) ( https://www.ncbi.nlm.nih.gov/geo/ ). After processing missing values with impute.knn function in CHAMP package, differences > 0.1 between tumor tissues and control samples (normal tissues or whole blood), standard deviation (SD) < 0.15 for tumor tissues and SD < 0.1 for control samples were nominated as differentially methylated sites (DMS). The adjacent ( < 250 bp) methylation sites were stitched into peaks. MCBs (methylation-correlated blocks) were defined as regions with more than or equal to 3 CpG sites within 100 bp and with DNA methylation sites correlation > 0.5. Results: In TCGA LUAD and LUSC, 658 and 551 DMS were identified between tumor and normal tissues; 999 and 912 DMS were generated by comparing tumor tissues and whole blood. The intersection of DMS located in the methylation canyon created a 203 kbp DNA methylation panel, containing 2521 DMS and 1417 methylation peaks. 37 MCBs were detected in tumor tissues of 30 lung cancer with DNA methylation panel sequencing. In term of 37 MCBs, 4 MCBs located in SHOX2, PTGER4, RASSF1A and ARL8B were identified with significant differences (p < 0.001) between plasma of 50 lung cancer patients and 50 healthy individuals. Furthermore, Real-time Quantitative PCR (qPCR) of the 4 MCB regions were performed in plasma of 30 lung cancer patients and 25 healthy individuals, and a cutoff of 0.414 distinguishing lung cancer from the healthy was calculated by the generalized linear model (GLM) based on the cycle threshold (CT) values of qPCR. The GLM incorporated the 4 MCBs showed robust performances in which the sensitivity, specificity and area under the curve (AUC) were 100%, 95% and 0.95, respectively. Postoperative plasma within 3 months were retrospectively collected from 32 lung cancer patients, and 30 patients with GLM scored less than 0.414 (Maximum: 0.327) had no recurrence within 1 year, while 2 patients with GLM scored more than 0.414 (0.480 and 0.557) were recurred within 1 year. Conclusions: The generalized linear model incorporated the 4 MCBs showed robust performances, and predicted the recurrence of early-stage lung cancer within 1 year after surgery.

Acute lymphoblastic leukemia (ALL) and its treatment continue to pose substantial risks. To understand ALL more deeply, the metabolome in fasting plasma of 27 ALL patients before and after high-dose methotrexate therapies (consolidation therapy) including methotrexate and 6-mercaptopurine (6-MP) was investigated. Plasma metabolites were analyzed using liquid chromatography–tandem mass spectrometry (LC–MS). Orthogonal projections to latent structures discriminant analysis and significance analysis of microarrays were used to evaluate the metabolic changes. Pathway enrichment and co-expression network analyses were performed to identify clusters of molecules, and 2826 metabolites were identified. Among them, 38 metabolites were identified by univariate analysis, and 7 metabolites that were altered by conditioning therapy were identified by multivariate analysis. The Kyoto Encyclopedia of Genes and Genomes (KEGG) database was used for pathway enrichment analysis. Among the enriched KEGG pathways, the 3 significantly altered metabolic pathways were pyrimidine metabolism; phenylalanine, tyrosine, and tryptophan biosynthesis; and phenylalanine metabolism. In addition, L-phenylalanine was significantly correlated with blood urea nitrogen (BUN), and palmitoylcarnitine was correlated with aspartate aminotransferase (AST). In summary, consolidation therapy significantly affected pyrimidine- and phenylalanine-associated metabolic pathways in pediatric ALL patients. These findings may provide an insight into the role of metabolic profiling in consolidation treatment and as a potential for pediatric ALL patients.

BACKGROUND:Effective targeted therapy for lung adenocarcinoma (LUAD), the number one cancer killer worldwide, continues to be a difficult problem because of the limitation of number of applicable patients and acquired resistance. Identifying more promising drug targets for LUAD treatment holds immense clinical significance. Recent studies have revealed that the U2 auxiliary factor (U2AF) homology motif kinase 1 (UHMK1) is a robust pro-oncogenic factor in many cancers. However, its biological functions and the underlying molecular mechanisms in LUAD have not been investigated. METHODS:The UHMK1 expression in LUAD cells and tissues was evaluated by bioinformatics analysis, immunohistochemistry (IHC), western blotting (WB), and real time quantitative polymerase chain reaction (RT-qPCR) assays. A series of gain- and loss-of-function experiments for UHMK1 were carried out to investigate its biological functions in LUAD in vitro and in vivo. The mechanisms underlying UHMK1's effects in LUAD were analyzed by transcriptome sequencing and WB assays. RESULTS:UHMK1 expression was aberrantly elevated in LUAD tumors and cell lines and positively correlated with tumor size and unfavorable patient prognosis. Functionally, UHMK1 displayed robust pro-oncogenic capacity in LUAD and mechanistically exerted its biological effects via the phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT)/mammalian target of rapamycin (mTOR) signaling pathway. CONCLUSION:UHMK1 is a potent oncogene in LUAD. Targeting UHMK1 may significantly improve the effect of LUAD treatment via inhibiting multiple biological ways of LUAD progression.

肺癌是严重威胁人类生命健康的疾病之一,目前其发病率和死亡率居所有癌症之首.microRNA-21(miR-21)作为一种致癌的miRNA(oncomiR),在多种实体瘤中的表达显著增加.miR-21在非小细胞肺癌(NSCLC)中可以通过改变各种靶分子的表达来调节癌细胞的增殖、侵袭、凋亡,降低遗传稳定性.本文系统性地综述了miR-21在NSCLC中的表达及其调控作用,从基因的表达、临床诊断、治疗与预后等方面探讨了miR-21在NSCLC发生发展中的作用、相关临床应用及潜在的应用价值.

[This corrects the article DOI: 10.3389/fcell.2021.771824.].

AbstractBackgroundAn increasing number of patients are being diagnosed with synchronous multiple primary lung cancer (SMPLC) with the popularization of lung cancer screening programs. However, a strategy for accurate location and suitable surgery therapy is still lacking. The present study aimed to explore the accuracy and feasibility of electromagnetic navigation bronchoscopy (ENB)‐guided thoracoscopic sublobectomy for stage IA SMPLC.MethodsPatients with SMPLC who underwent ENB‐guided sublobectomy from January 2020 to June 2022 were enrolled in this study. The analysis of localization accuracy of ENB and surgical outcome was conducted.ResultsOverall, 138 patients with 353 malignant nodules were enrolled. The tumor size was 0.7 cm (range from 0.5 to 1.1 cm). ENB localization was performed on 162 nodules, and a customized scoring system was developed to evaluate localization accuracy. The success rate of localization was 98.3% (178/181). Notably, localization accuracy was positively correlated with bronchial signs (p < 0.01) and negatively correlated with the distance from the nodule to the pleura (p = 0.02). All nodules were completely resected. Operation time, drainage volume on the third postoperative day, and catheter time were significantly correlated with the resected lesion numbers (p = 0.009, p = 0.004, and p = 0.01, respectively).ConclusionsENB‐guided uniportal video‐assisted thoracoscopic sublobectomy provides accurate preoperative localization and avoids unnecessary lung resection of patients with stage IA SMPLC. However, complete resection of multilocation nodules (more than four lesions) increases the risk of postoperative complications. A new combined treatment strategy for SMPLC should be explored.

BACKGROUND:Circular RNA (circRNA) has become a new focus in the field of tumor biology research in recent years. Many circRNAs have been showed to play an important role in the progression of lung adenocarcinoma (LUAD). In this work, we studied the oncological role of hsa-circ-000881 in LUAD and attempted to explore the related mechanism.METHODS:The relative expressions of hsa-circ-000881, miR-665, and PRICKLE2 were detected by RT-qPCR or western blot. Functional assays were conducted to analyze the role of hsa-circ-000881 in the proliferation, migration, and invasion of LUAD cells. A luciferase reporter assay was performed to verify whether hsa-circ-000881, miR-665, and PRICKLE2 interact with each other.RESULTS:Circ-000881 was remarkably downregulated in LUAD. Overexpression of circ-000881 attenuated cell growth, migration, and invasion, whereas its knockdown enhanced the malignancy of LUAD cells. The results of luciferase reporter assay and bioinformatics analysis confirmed that circ-000881 served as a sponge for miR-665, and PRICKLE2 was a direct target of miR-665.Overexpression of miR-665 or silencing of PRICKLE2 abolished circ-000881-mediated inhibition of malignant tumor behavior in LUAD cells.CONCLUSIONS:Circ-000881 has inhibitory effects on LUAD via a miR-665/PRICKLE2 axis, suggesting that circ-000881 may be an underlying therapeutic target for LUAD.

Background: Lung adenocarcinoma (LUAD) is the major form of lung cancer that presents a major peril to public health. Owing to the high rates of morbidity, mortality and chemoresistance, it is necessary to develop more effective therapeutic targets of LUAD. Mitochondrial fission regulator 1 (MTFR1) affects the occurrence and development of some diseases by regulating mitochondrial dynamics and is dysregulated in LUAD. However, the functions and molecular mechanisms of MTFR1 in LUAD have not been investigated.Methods: Immunohistochemical (IHC) analysis, real-time quantitative polymerase chain reaction (RT-qPCR), bioinformatic analysis and western blot (WB) were performed to assess the expression of MTFR1 at both protein and mRNA levels. The biological functions of MTFR1 in LUAD cells were assessed based on various in vivo and in vitro experiments. The dual-luciferase reporter assay and some rescue experiments were performed to evaluate the underlying mechanism of MTFR1 in LUAD.Results: MTFR1 was upregulated in LUAD cells and tissues and correlated with dismal clinicopathologic features and a worse prognosis of patients with LUAD. Functionally, MTFR1 overexpression stimulated the proliferation, invasion, migration and glycolytic capacity and impeded the apoptosis of LUAD cells; however, opposite results were obtained when MTFR1 expression was knocked down. MTFR1, which was directly targeted by miR-29c-3p, may exert its biological functions through the AMPK/mTOR signalling pathway.Conclusion: MTFR1 promotes the progression of LUAD. Therefore, targeting MTFR1 can offer an effective therapeutic strategy for LUAD treatment.

Background: Many evidences proved important roles of microRNAs in the occurrence and development of non-small cell lung cancer (NSCLC). This work aimed to explore the effects and regulating mechanism of microRNA-665 (miR-665) in NSCLC. Methods: Here, quantitative real time PCR was used to evaluate the expression level of miR-665 in NSCLC tissues and several cell lines (H520, H1299, PC9, H358, and A549). Then, MTT, apoptosis assays and wound-healing assays were performed to study miR-665 functions on cell proliferation, apoptosis and migration. Based on bioinformatics analysis, nemo-like kinase (NLK) was predicted as a potential target of miR-665 and then a series of experiments were performed to explore miR-665 mechanism.Results: In NSCLC tissues and several cell lines, miR-665 was up-regulated and cell experiments displayed that it had abilities to promote cell proliferation and migration, and inhibit cell apoptosis. MiR-665 over-expression could reduce NLK mRNA and protein level by binding to 3′-untranslated region of NLK, which suggested that miR-665 could regulate NLK expression. Further analysis found that there was an inverse correlation f between miR-665 and NLK in NSCLC. Moreover, low-level NLK possessed same effects on cells as miR-665 over-expression, and its knockdown could partly recovery the miR-665-mediated cell proliferation, migration and anti-apoptosis, indicating that miR-665 might regulate biological behaviors of NSCLC cells via targeting NLK. Conclusion: MiR-665 probably promoted NSCLC occurrence and progression via targeting NLK, which might be a hopeful biomarker for NSCLC diagnosis or a target for treatment.

Objective. This Work is aimed at exploring the effect of microRNA (MiR)-608 on the function of nonsmall cell lung cancer (NSCLC) A549 cells and related mechanisms. Methods. Blood samples of 106 NSCLC patients (experimental group) as well as 124 normal people (control group) were selected for relevant investigation. Polymerase chain reaction (PCR) as well as DNA sequencing was used to determine the genotyping of the MiR-608 rs4919510 polymorphism. MiR-608 expression in cells was detected by real-time PCR technology. Western blotting was used to detect changes in protein levels. NSCLC tissues as well as adjacent tissues were explored in 33 patients undergoing surgery. Results. MiR-608 rs4919510 does not influence the incidence of NSCLC patients. In addition, MiR-608 expression was downregulated in the tumor tissue of NSCLC patients, while the transcription factor activating enhancer-binding protein 4 (TFAP4) expression was upregulated. MiR-608 promotes DOX- (Doxorubicin-) induced apoptosis by negatively regulating TFAP4 expression in NSCLC tissue. TFAP4 can significantly inhibit the migration of A549 cells. Conclusion. The findings in this investigation can contribute to the effective treatment of NSCLC patients. Also, the investigation can provide some theoretical support for the application of new targets for NSCLC treatment.

目的 在倾向性评分匹配配对良好的情况下,比较机器人与胸腔镜在肺癌手术治疗中的围手术期安全性与短期疗效.方法 回顾性分析2020年8月至2020年10月期间,山东大学齐鲁医院胸外科田辉教授肺外科团队因原发性肺癌行肺叶或亚肺叶切除+肺门纵隔淋巴结清扫或采样术的286例患者的临床资料.其中,130例行达芬奇机器人辅助胸外科(RATS)肺切除术,为RATS组;156例行电视胸腔镜辅助胸外科(VATS)肺切除术,为VATS组.采用倾向性评分匹配方法进行混杂因素校正,比较匹配后两组病例的围手术期结果.结果 倾向性评分匹配分析后,每组88例配对成功.对两组病例的围手术期临床资料行统计学分析,发现RATS组手术时间略长于VATS组,但差异无统计学意义(P=0.625).RATS组术中出血量较VATS组更少(P<0.001).RATS组淋巴结清扫站数(P<0.001)及清扫个数(P=0.031)均高于VATS组;RATS组住院费用较VATS组高,差异有统计学意义(P<0.001).术后第1～3天疼痛数字评分(NRS评分)差异有统计学意义(P<0.001),RATS组术后第1～3天NRS评分更高.两组淋巴结升期率、术后第1～3天引流量、术后全部拔管时间、术后住院天数、术后并发症差异无统计学意义(P>0.05).结论 在可切除肺癌手术治疗上,RATS与VATS的围手术期安全性及短期疗效相似.此外,RATS在术中出血量、淋巴结清扫彻底性上存在优势,缺点是增加了住院总费用,潜在增加了术后疼痛.

加速康复外科(enhanced recovery after surgery,ERAS)理念由丹麦哥本哈根大学的Kehlet[1-2]首次提出.2007年,由黎介寿院士将ERAS理念引入我国[3].目前已被广泛的应用于临床外科各个专业.在2018年世界十大医疗创新成果中,ERAS位居第8位.ERAS是以患者为中心,在外科、麻醉、护理、营养等多学科团队协作的基础上,通过一系列具有循证医学证据的围手术期优化措施的综合应用,以减少手术患者的生理及心理的创伤应激反应,达到加速康复的目的[4].

在全世界范围内,肺癌的发病率及病死率均位居所有恶性肿瘤的第一位[1].虽然近年化疗、放疗、靶向治疗或免疫治疗等取得了长足的进展,但其仅能够一定程度改善肺癌患者的预后,目前外科手术治疗仍是公认的治疗早中期肺癌的最有效方法[2].随着电视胸腔镜手术(video-assisted thoracic surgery,VATS)在肺癌外科治疗领域的广泛应用及不断发展,研究表明VATS相比传统开放手术,患者的术后疼痛更轻、住院时间更短、生活质量更高,但其并不能有效改善肺癌患者的长期预后(包括无病生存期及总生存期等);与此同时,研究发现规范化及标准化的手术质量控制尤为重要,其可显著提高患者的围手术期疗效并明显改善肺癌患者的预后[3-5].