Vitamin D supplementation for prevention of acute respiratory infections in older adults: A systematic review and meta-analysis.

BACKGROUND: Acute respiratory infections (ARIs) have a substantial impact on morbidity, healthcare utilization, and functional decline among older adults. Therefore, we systematically reviewed evidence from randomized controlled trials (RCTs) to evaluate the efficacy and safety of vitamin D supplementation in preventing ARIs in older adults. METHODS: PubMed, Embase, the Cochrane Library, and ClinicalTrials.gov were searched until 1 February 2024. RCTs evaluating the use of vitamin D supplements to protect older adults from ARIs were included. Two reviewers independently screened papers, extracted the data and assessed the risk of bias. Data were summarised as relative risks (RRs) or odds ratios (ORs) with corresponding 95% confidence intervals (CIs). Random effects meta-analyses were used to synthesise the results. GRADE was used to evaluate the quality of evidence. All the analysis were performed with Stata version 17. RESULTS: Twelve trials (41552 participants) were included in the meta-analysis. It showed that vitamin D supplementation probably does not reduce the incidence of ARIs (RR, 0.99; 95% CI, 0.97-1.02, I2 = 0%; moderate certainty). No significant effect of vitamin D supplementation on the risk of ARI was observed for any of the subgroups defined by baseline 25(OH)D concentration, control treatments, dose frequency, study duration, and participants' condition. However, there was a possibility, although not statistically significant, that vitamin D may reduce the risk of ARI in patients with a baseline 25(OH)D concentration <50 nmol/L (OR, 0.90; 95% CI, 0.79-1.04, I2 = 14.7%). Additionally, vitamin D supplements might result in little to no difference in death due to any cause, any adverse event, hypercalcinemia, and kidney stones. CONCLUSIONS: Vitamin D supplementation among older adults probably results in little to no difference in the incidence of ARIs. However, further evidence is needed, particularly for individuals with vitamin D deficiency and populations residing in low and middle income countries. TRIAL REGISTRATION: This study was registered on PROSPERO (CRD42023451265).

Efficacy and safety of immune checkpoint inhibitors and targeted therapies in resected melanoma: a systematic review and network meta-analysis.

Background: Multiple immune checkpoint inhibitors (ICIs) and targeted therapies have been widely used as adjuvant treatments for high-risk resected melanoma, with unclear comparative efficacy and safety. Methods: PubMed, Embase, the Cochrane Library, and ClinicalTrials.gov were searched from database inception until 6 June 2023. We included RCTs that assess adjuvant ICIs or targeted therapies in high-risk resected melanoma. Frequentist random-effect network meta-analyses (NMA) were performed. The primary outcome was recurrence-free survival (RFS). Results: Eleven trials including 10,712 patients and comparing 10 treatments (nivolumab [Nivo], ipilimumab 3 mg/kg [Ipi3], Ipi10, pembrolizumab [Pemb], vemurafenib [Vemu], bevacizumab [Beva], Nivo + Ipi1, Nivo + Ipi3, dabrafenib plus trametinib [Dab + Tram], and placebo/observation [Pla/Obs]) were included. NMA showed that all treatments showed RFS benefit over placebo/observation except Ipi3 (hazard ratio [HR], 0.78; 95% CI, 0.58-1.05). Combination therapy of Nivo + Ipi3 was the most effective treatment, which significantly improved RFS compared with other treatments. NMA also showed that all treatments were associated with an increased risk of grade 3-5 adverse events over placebo/observation except Nivo (HR, 1.25; 95% CI, 0.87-1.80). NMA suggested that Nivo and Pemb were the two safest treatments except for placebo/observation. Although three combination therapies ranked as the top three in terms of RFS, they did not show significant overall survival benefits compared to monotherapies including Pemb, Nivo, Ipi3, and Ipi10. Conclusion: In this NMA, adjuvant Nivo and Pemb are the preferred options in patients with resected melanoma considering the benefits and harms. Combination therapy of Nivo + Ipi3 may be a promising strategy, but more evidence from phase 3 trials is needed. Systematic Review Registration: https://www.crd.york.ac.uk/PROSPERO/display_record.php?RecordID=438667, PROSPERO (CRD42023438667).

Apoptosis-Related Signature Predicts Prognosis and Immune Microenvironment Infiltration in Lung Adenocarcinoma.

Lung adenocarcinoma (LUAD), a malignancy with high incidence and mortality rates worldwide, contains multiple genomic and epigenomic abnormalities. And the useful tumor markers associated with these abnormalities need further investigation. Whereas apoptosis is a form of programmed cell death, the expression of apoptosis-related genes in LUAD and its relationship with prognosis is unclear. In the present study, we identified 64 differentially expressed apoptosis-related genes (DEARGs) that were differentially expressed between LUAD tissue and normal lung tissue. Based on these DEARGs, all LUAD cases were classified into two subtypes using The Cancer Genome Atlas (TCGA) cohort to assess the prognostic value of apoptosis-related genes for survival. An 11-gene signature was established by applying the Least Absolute Shrinkage and Selection Operator (LASSO) Cox regression method to construct a multigene prediction model and classify all LUAD patients in the TCGA cohort into high or low AS-score groups. Patients in the low AS-score group had significantly higher survival and prognosis than those in the high AS-score group. Taking the median risk score of the AS-score, LUAD patients in the GSE68465 cohort were divided into two risk groups, low and high. The overall survival (OS) time was longer in the low AS-score group. Combined with clinical characteristics, the AS-score was an independent predictor of LUAD patients. Gene ontology (GO) and Kyoto Encylopedia of Genes and Genomes (KEGG) analyses showed that the differential genes between the two groups were mainly enriched in cellular immunity. Further analysis revealed higher immune checkpoint protein expression and higher tumor mutational burden (TMB) in the high AS-score group, suggesting better efficacy of immunotherapy in the high AS-score group than the low AS-score group. And the high AS-score group was better in chemotherapy and targeted therapy efficiency. In conclusion, the AS-score constructed based on apoptosis-related genes can predict the prognosis of LUAD patients and provide some guidance for the antitumor treatment of LUAD patients.

Long non-coding RNA SNHG17 promotes lung adenocarcinoma progression by targeting the microRNA-193a-5p/NETO2 axis.

Long non-coding RNAs (lncRNAs) play vital roles in human cancers. It has been reported that lncRNA SNHG17 expression is dysregulated in different types of cancer and involved in cancer progression. However, the role of SNHG17 in lung adenocarcinoma (LUAD) remains unclear. The present study aimed to investigate the role of SNHG17 in LUAD. Reverse transcription-quantitative (RT-q) PCR analysis was performed to detect SNHG17 expression in LUAD tissues and cells. The effects of SNHG17 on cancer cell migration, invasion, proliferation and epithelial-to-mesenchymal transition (EMT) were assessed via Transwell, MTT and western blot assays, respectively. The interactions between SNHG17 and microRNA (miRNA/miR)-193a-5p, miR-193a-5p and neuropilin and tolloid-like 2 (NETO2) were assessed via the dual-luciferase reporter assay. NETO2 expression and its potential role in LUAD were analyzed via RT-qPCR analysis and the UALCAN database. The results demonstrated that SNHG17 expression was significantly upregulated in LUAD tissues and cells, and high SNHG17 expression was associated with tumor-node-metastasis stage and poor prognosis of patients with LUAD. SNHG17 knockdown inhibited cell migration, invasion, proliferation and the EMT process. In addition, the results revealed that SNHG17 functions as a competing endogenous RNA of miR-193a-5p. The results of the dual-luciferase reporter assay confirmed that miR-193a-5p can directly target SNHG17. NETO2 was also predicted as a target protein of miR-193a-5p, which was confirmed via the dual-luciferase reporter assay. The roles of NETO2 knockdown in cancer cells were rescued following transfection with miR-193a-5p inhibitor or overexpression of SNHG17. Notably, high NETO2 expression was associated with poor prognosis of patients with LUAD. Bioinformatics analysis demonstrated that the promoter methylation level of NETO2 decreased in LUAD. Taken together, the results of the present study suggest that SNHG17 expression is upregulated in LUAD tissues and cells, and SNHG17 exerts tumor promoting effect by targeting the miR-193a-5p/NETO2 axis.

[Gene Detection Analysis of Thalassemia in 2 494 Cases of Childbearing Couples in Haikou City].

OBJECTIVE: To observe the genotypes and composition ratio of thalassemia in couples of reproductive age, and provide a reference for the prevention and control of thalassemia in Haikou. METHODS: Gene diagnosis was performed in 2 494 subjects who were screened for thalassemia before marriage or prenatal by cross-breakpoint PCR, PCR-reverse dot hybridization, and PCR-electrophoresis. RESULTS: A total of 1 037 thalassemia gene carriers were detected in 2 494 samples, with a detection rate of 41.57%, of which 75.02% was α-thalassemia, 18.61% was ß-thalassemia, and 6.36% was α-ß complex thalassemia. There were 778 cases of α-thalassemia, mainly of deletion type, accounting for 76.99% (599/778). Twenty genotypes were detected, the highest three was --SEA/αα (33.42%, 260/778), -α3.7/αα(23.91%, 186/778), and -α4.2/αα(19.02%, 148/778), respectively. A rare HKαα/-α3.7 was detected, who immigrated from other province. There were 193 cases of ß-thalassemia, all of them were light (ß0/ßA or ß+/ßA). Eight genotypes were detected, the highest two was 41-42M/N (74.61%, 144/193) and 654M/N (10.36%, 20/193), respectively. There were 66 cases of α-ß compound type of thalassemia, 15 genotypes were detected, the highest three was ααWS/αα complex 41-42M/N (28.79%, 19/66), -α3.7/αα complex 41-42M/N, and -α4.2/αα complex 41-42M/N (16.67%, 11/66 for both). CONCLUSION: In Haikou city, the gene carrying rate of thalassemia is very high, and the genotype distribution is different from other cities in Hainan Province, attention should be paid to the impact of population inflow on the frequency spectrum change of local thalassemia gene.

Folic Acid-Conjugated CuFeSe2 Nanoparticles for Targeted T2-Weighted Magnetic Resonance Imaging and Computed Tomography of Tumors In Vivo.

BACKGROUND: Development of new long-circulating contrast agents for computed tomography (CT) and magnetic resonance imaging (MRI) of different biological systems still remains a great challenge. Here, we report the synthesis of folic acid (FA)-targeted CuFeSe2 nano-contrast agent for CT and MRI imaging in vitro and in vivo. METHODS AND RESULTS: In our study, CuFeSe2 was fabricated through a facile and green aqueous reaction and then further aminated through silanization. The amine-functionalized CuFeSe2-NH2 nanoparticles enable the covalent conjugation of folate-conjugated polyethylene glycol (FA-PEG-COOH) as a targeting ligand onto their surface, which could improve the dispersion and endue the targetability of nanoparticles, respectively. The formed multifunctional CuFeSe2-PEG-FA nanoparticles were characterized via different techniques, which exhibited outstanding dispersion, good biocompatibility and excellent FA-targeted capability. Meanwhile, the nanoparticles were quite safe in the given concentration range as confirmed by in vitro and in vivo toxicity assay. Importantly, CuFeSe2-PEG-FA nanoparticles were successfully applied in CT/MRI dual-modality imaging in vitro and in vivo, which showed a better imaging performance and targeted capability. CONCLUSION: Therefore, the constructed CuFeSe2-PEG-FA nanoparticles have a great potential as an efficient contrast agent for dual-modality imaging of different biological systems.

Feasibility of magnetic resonance imaging-based radiomics features for preoperative prediction of extrahepatic cholangiocarcinoma stage.

AIM: The aim of this study is to develop and test radiomics models based on magnetic resonance imaging (MRI) to preoperatively and respectively predict the T stage, perineural invasion, and microvascular invasion of extrahepatic cholangiocarcinoma (eCCA) through a non-invasive approach. METHODS: This research included 101 eCCA patients (29-83 years; 45 females and 56 males) between August 2011 and December 2019. Radiomics features were retrospectively extracted from T1-weighted imaging, T2-weighted imaging, diffusion-weighted imaging, and apparent diffusion coefficient map using MaZda software. The region of interest was manually delineated in the largest section on four MRI images as ground truth while keeping 1-2 mm margin to tumor border, respectively. Pretreatment, dimension reduction method, and classifiers were used to establish radiomics signatures for assessing three pathological characteristics of eCCA. Finally, independent training and testing datasets were used to assess radiomics signature performance based on receiver operating characteristic curve analysis, accuracy, precision, sensitivity, and specificity. RESULTS: This study extracted 1208 radiomics features from four MRI images of each patient. The best performing radiomics signatures for assessing the T stage, perineural invasion, and microvascular invasion were respectively produced by L1_normalization + linear discriminant analysis (LDA) + logistic regression, Box_Cox transformer + LDA + K-nearest neighbor, and L2_normalization + LDA + AdaBoost. The area under the curve values of the radiomics signatures for predicting the training and testing cohorts in each subgroup were respectively 1 and 0.962 (T stage), 1 and 1 (both perineural invasion and microvascular invasion). CONCLUSION: These proposed radiomic models based on MR images had powerful performance and high potential in predicting T stage, perineural, and microvascular invasion of eCCA. REPORTING GUIDELINES/RESEARCH DESIGN: Prognostic study.

Circular RNA circAFF2 accelerates gastric cancer development by activating miR-6894-5p and regulating ANTXR 1 expression.

BACKGROUND: Circular RNAs (circRNAs) contain a new class of non-coding RNAs that play an important role in adjusting biological function and gene expression. But the function of circRNAs in gastric cancer remains unclear. In the present research, we explored the functions of circular RNA AFF2(circAFF2, hsa_circ_0001947) in gastric cancer cells and an animal model of gastric cancer. METHODS: The expression of circAFF2, microRNA-6894-5p (miR-6894-5p), and Anthrax toxin receptor 1 (ANTXR 1) were determined by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Cell counting kit 8 (CCK-8) and transwell assays were used to analyze the knockdown effects of circAFF2, miR-6894-5p, and overexpression of ANTXR 1 on cell proliferation, migration, and invasion abilities. Binding interactions between, circAFF2 and miR-6894-5p and between, miR-6894-5p and ANTXR 1 were detected by Dual-luciferase reporter assays. Levels of protein expression were analyzed by Western blotting. Tumor models were established by subcutaneous injection of tumor cells in nude mice. RESULT: The result showed that circAFF2 expression was significantly increased in gastric cancer cell lines and tissues. The knockdown of circAFF2 dramatically suppressed the cell migration, invasion and proliferation of gastric cancer cells. In vivo studies showed that knockdown of circAFF2 delayed tumor growth. Furthermore, we revealed that circAFF2 functioned as a sponge to absorb miR-6984-5p and elevated the expression of ANTXR 1. CONCLUSION: CircAFF2 acts as an oncogene in gastric cancer and exerts its effects via miR-6894-5p/ANTXR 1 signaling.

Dexamethasone Upregulates the Expression of Aquaporin4 by Increasing SUMOylation in A549 Cells.

Dexamethasone can alleviate the severity of bronchial and alveolar edema and therefore is widely applied in the treatment of various exudative diseases including pulmonary edema. However, the effectiveness of dexamethasone is still being questioned and its mechanism is not fully understood. Aquaporins (AQPs) are mainly responsible for the transmembrane transport of water, which is tightly associated with pulmonary edema. Small ubiquitin-like modifiers (SUMOs) are considered to play a protective role in some pathological conditions. In this study, we demonstrated that dexamethasone can upregulate the expression of AQPs in A549 cells by inducing SUMOylation. We found that a low dose of dexamethasone significantly upregulated the levels of SUMOylation and AQP expression in A549 cells, accompanied by a translocation of SUMOs from the cytoplasm to the nucleus. We also explored the possible relation between SUMOylation and AQPs. Knockdown of SUMO2/3 by RNA interference decreased the level of AQP4 in A549 cells after dexamethasone stimulation. Together, our findings demonstrated that AQP4 expression was upregulated in A549 cells exposed to dexamethasone, and SUMOylation may participate in the regulation of AQP4.

Recombinant Newcastle disease virus expressing human IFN-λ1 (rL-hIFN-λ1) inhibits lung cancer migration through repolarizating macrophage from M2 to M1 phenotype.

BACKGROUND: Tumor-associated macrophages (TAMs) are frequently infiltrated in tumor microenvironment and promote tumor progression. Lung cancer development largely depends upon the essential contributions from the TAMs which generally polarize into M2 TAMs and produce abundant anti-inflammatory factors and facilitate tumor development. The recombinant Newcastle disease virus expressing human IFN-λ1 (rL-hIFN-λ1) could regulate Th1/Th2 immune response to produce anti-tumor microenvironment. However, the interaction between rL-hIFN-λ1 and macrophages polarization remains unclear. METHODS: The THP-1 cells were used to construct the THP-1-M0, THP-1-M1, THP-1-M2 and THP-1-rL-hIFN-λ1 macrophage models. qRT-PCR and Immunofluorescence were used to detect the polarization phenotype of macrophage polarized by rL-hIFN-λ1. The inhibitory properties of THP-rL-hIFN-λ1 on A549 cells and H446 cells were determined by a Clonogenic assay, as well as scratch migration assays and Transwell were used to explore the capability of migration. Furthermore, the M1/M2 infiltration density in different clinical stages of lung cancer tissues were examined. RESULTS: It was showed that rL-hIFN-λ1 could induce normal macrophages to differentiate into THP-1-M1 macrophages. Meanwhile, rL-hIFN-λ1 could also direct THP-1-M2 macrophages polarization into THP-1-M1 macrophages. Supernatants from rL-hIFN-λl induced macrophages inhibited colony formation, migration and invasion of lung cancer cells in vitro which was similar to THP-1-M1 macrophages. Moreover, analysis of clinical tumor tissues indicated that M1-type macrophages decreased gradually with the development of the clinical stage of lung cancer. CONCLUSIONS: Therefore, rL-hIFN-λl induced significant suppression of primary lung tumor growth and spontaneous lung metastases through regulating macrophages function, and it was expected to become a new biological therapy for lung cancer.

Recombinant virus expressing hIFN-λ1 (rL-hIFN-λ1) has important effects on endoplasmic reticulum stress, autophagy and apoptosis in small cell lung cancer.

BACKGROUND: Small cell lung cancer (SCLC) is an aggressive tumor with a poor prognosis. Human IFN-λ1 (IL-29), belonging to the type III IFN family, captured increasing attention recently due to its crucial role in developing tumors. Recent studies have revealed that the recombinant Newcastle Disease Virus (NDV) expressing human IFN-λ1 (rL-hIFN-λ1) plays a critical role in the development of tumors. However, the role of rL-hIFN-λ1 in SCLC is still unknown. METHODS: We determined the concentration of the virus intervention, followed by successfully infection in virus. We also investigated the effects of rL-hIFN-λ1 on endoplasmic reticulum stress (ERS), apoptosis and autophagy in H446 cells, and explored the interaction among the three. RESULTS: We found that the ERS, autophagy and apoptosis related proteins were significantly upregulated after infected with rL-hIFN-λ1 or NDV. In addition, both 4-phenylbutyric acid (4-PBA) or 3-Methyladenine (3-MA) could downregulate the expression of related proteins which increased by rL-hIFN-λ1. Furthermore, we found that both B-cell lymphoma-2 (BCL-2) knockdown or Rapamycin (Rapa) could increase ERS, autophagy and apoptosis. CONCLUSIONS: Our findings suggest that rL-hIFN-λ1 can induce ERS, autophagy and apoptosis in SCLC H446 cells, particularly, autophagy plays an important role during this process. Furthermore, rL-hIFN-λ1 might provide a potential biological treatment target for lung cancer treatment.

Next-generation sequencing reveals hsa_circ_0058092 being a potential oncogene candidate involved in gastric cancer.

Gastric cancer is a serious problem for human health. As part of noncoding RNA, circular RNA (circRNA) plays a key role in the occurrence and development of malignant tumor. We used next generation sequencing technology to detect circRNA expression profiles in 5 paired human gastric cancer tissues. Then, bioinformatics analysis was carried out to analyze the function of dysregulated circRNAs. Hsa_circ_0058092 was selected as the object of follow-up analysis. After using the Cistrome DB dataset the data was used to predict specific transcription factors of hsa_circ_0058092. The relationship between hsa_circ_0058092 and PODXL was further validated using RT-PCR and immunohistochemical techniques. Survival data were collected using a Kaplan-Meier analysis of hsa_circ_0058092. We identified 319 aberrantly expressed circRNAs, Hsa_circ_0058092 was selected for our studies. Functional analysis of hsa_circ_0058092 revealed that it was related to metabolic processes. The prediction results suggested that hsa_circ_0058092 has a relationship with hsa-miR-4269 which could specifically bind to the PODXL sequence. Transcription factor CEBPB may regulate the transcription process of hsa_circ_0058092. The expression of hsa_circ_0058092 was positively correlated with PODXL expression. Immunohistochemical analysis of PODXL showed that the expression of PODXL protein in cancer tissues is higher than that in adjacent tissues. Kaplan-Meier analysis suggested that hsa_circ_0058092 was associated with survival of gastric cancer patients. All of these results showed that hsa_circ_0058092 was a potential oncogene.

Migration of gastric cancer is suppressed by recombinant Newcastle disease virus (rL-RVG) via regulating α7-nicotinic acetylcholine receptors/ERK- EMT.

BACKGROUND: Nicotinic acetylcholine receptors (nAChRs) have been reported to be overexpressed in malignancies in humans and is associated with tumorigenesis and cell migration. In previous studies of gastric cancer, alpha7 nicotinic acetylcholine receptor (α7-nAChR) overexpression leads to epithelial-mesenchymal transition (EMT) and promotes the migration of gastric cancer cells. Recombinant avirulent LaSota strain of Newcastle disease virus (NDV) expressing the rabies virus glycoprotein (rL-RVG) may promote apoptosis of gastric cancer cells and reduces the migration of lung cancer metastasis. However, whether rL-RVG inhibits migration of gastric cancer cells and what the underlying functional mechanism is remains unknown. METHODS: The gastric cancer cell lines BGC and SGC were randomly divided into 3 groups: rL-RVG, NDV and Phosphate Buffered Solution (PBS) control groups. Furthermore,we adopted ACB and MLA,α7nAChR-siRNA for the overexpression and silencing of α7-nAChR.Corynoxenine was used for inhibiting the MEK-ERK pathway. Western blot, Immunofluoresce,cell proliferation assays,cell migration analyses through wound-healing assays and Transwell assays were used to explore the underlying mechanisms. A mouse xenograft model was used to investigate the effects of rL-RVG,NDV on tumor growth. RESULTS: In this study, our findings demonstrate that rL-RVG suppressed the migration of gastric cancer cells and reduced EMT via α7-nAChR in vitro. Furthermore rL-RVG decreased the phosphorylation levels of the MEK/ERK signaling pathway such as down-regulating the expression of P-MEK and P-ERK. Additionally, rL-RVG also reduced the expression level of mesenchymal markers N-cadherin and Vimentin and enhanced the expression of the epithelial marker E-cadherin. Lastly, rL-RVG inhibited nicotinic acetylcholine receptors (nAChRs) to suppress cell migration and epithelial to mesenchymal transition (EMT) in gastric cell. We also found that rL-RVG suppresses the growth of gastric cancer subcutaneous tumor cells in vivo. CONCLUSION: rL-RVG inhibits α7-nAChR-MEK/ERK-EMT to suppress migration of gastric cancer cells.

LaSota Strain Expressing The Rabies Virus Glycoprotein (rL-RVG) Suppresses Gastric Cancer by Inhibiting the Alpha 7 Nicotinic Acetylcholine Receptor (α7 nAChR)/Phosphoinositide 3-Kinase (PI3K)/AKT Pathway.

BACKGROUND The recombinant avirulent Newcastle disease virus (NDV) LaSota strain expressing the rabies virus glycoprotein (rL-RVG) can induce much greater apoptosis than can NDV in gastric carcinoma cells, but the mechanisms involved remains unclear. MATERIAL AND METHODS The 2 gastric carcinoma cell lines were divided into the rL-RVG group, the NDV group, and the PBS group. MTT assay was used to detect and analyze cell viability. siRNA for alpha7-nAChR, alpha7-nAChR antagonist, or alpha7-nAChR agonist, AKT antagonist, and p-AKT agonist were used for pretreatment. The protein expressions of RVG, NDV, alpha7-nAChR, cleaved caspase-3, p-AKT, PI3K, Bcl-2, and Bax proteins were detected by Western blot assay. Immunofluorescence was used to detect expressions of alpha7-nAChR proteins. Light microscopy, flow cytometry, and TUNEL assay were used to assess apoptosis. RESULTS The results showed that 2 virus concentrations over 10³ dilution caused greater cell proliferation inhibition. rL-RVG treatment increased the expression of alpha7-nAChR, cleaved caspase-3, and Bax protein but decreased the expression of p-AKT, PI3K, and Bcl-2 protein. When the groups were pretreated with alpha7-nAChR antagonist, the alpha7-nAChR, cleaved caspase-3, and Bax protein expression increased, but the expression of p-AKT, PI3K, and Bcl-2 protein was clearly decreased. However, the results in the alpha7-nAChR agonist group were the opposite. When treated with the AKT antagonist, the result was the same as in the rL-RVG treatment group. The result in the AKT agonist group was the opposite of that in the AKT antagonist group. Compared with the NDV group, the results of light microscopy, FCM, and TUNEL assay showed that alpha7-nAChR antagonist significantly affected the apoptosis of gastric cancer cells in the rL-RVG group. CONCLUSIONS rL-RVG leads to much greater apoptosis through the alpha7-nAChR/PI3K/AKT pathway.

RNA-Seq profiling of circular RNAs and potential function of hsa_circ_0002360 in human lung adenocarcinom.

CircRNAs have been identified play a key role in various different types of cancer. However, their role in lung adenocarcinoma remains unclear. In this study, we explored the specific circular transcriptome and characterized the circRNA expression profiles of five paired lung adenocarcinoma (LAC) tissues relative to adjacent normal tissues from LAC patients using next-generation sequencing (NGS). To illuminate circRNAs function, their gene targets were initially predicted before using, Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) analyses to further analyse the associated significant cell signalling pathways and functions. The potential interactions between circRNA-miRNA-mRNA were also investigated. Additionally, qRT-PCR assay, Western blot and Immunohistochemistry were performed to validate the differential expression of circRNA, microRNA and mRNA in the LAC group in comparison to the control group. Two-hundred-eighty-five dysregulated circular transcripts were found in LAC tissues, among which 102 and 183 were either up or down regulated, respectively. Our biological analysis suggested that the host genes of differentially expressed circRNAs targeted to cancer-related processes and mechanisms. The interaction maps of the circRNA-miRNA-target gene were constructed using Cytoscope. In further study, hsa_circ_0002360 was found to be the most significantly overexpressed circRNA in LAC tissues by interacting with miRNA and its corresponding mRNA. Our results showed that hsa_circ_0002360 was aberrantly and abundantly expressed and implicated in the development of LAC, suggesting a valuable therapeutic target for LAC treatment.

Recombinant Newcastle disease virus expressing human IFN-λ1 (rL-hIFN-λ1)-induced apoptosis of A549 cells is connected to endoplasmic reticulum stress pathways.

BACKGROUND: IFN-λs are a kind of cytokine with anti-tumor, immunomodulatory, and anti-proliferative activity. Recent studies have shown that the recombinant Newcastle disease virus expresses human IFN-λ1 (rL-hIFN-λ1), which plays a role in gastric cancer cell apoptosis. Endoplasmic reticulum stress (ERS) induces autophagy and apoptosis in tumor cells. In this study, we explored the relationship between ERS and rL-hIFN-λ1-induced apoptosis of lung adenocarcinoma A549 cells and its underlying mechanism. METHODS: First, we investigated the effect of rL-hIFN-λ1 on cellular proliferation, migration, and proteins associated with ERS, autophagy, and apoptosis of A549. Second, after administration of the ERS inhibitor, the associated proteins induced by rL-hIFN-λ1 were detected. Finally, a subcutaneous mouse model was used to examine the effect of rL-hIFN-λ1 on tumor growth and the ERS and apoptosis associated proteins in tumor tissues. RESULTS: The results showed that the proliferation and migration of A549 cells, and tumor tissue growth were significantly inhibited and the ERS, autophagy, and apoptosis associated proteins were upregulated in the experimental group. Additionally, both 4-PBA and knockdown of PERK or CHOP reduced the levels of rL-hIFN-λ1-induced autophagy and apoptosis-associated proteins. BCL-2 knockdown caused autophagy and apoptosis associated protein upregulation. CONCLUSIONS: In summary, rL-hIFN-λ1 inhibited cell proliferation and activated ERS, autophagy, and apoptosis in A549 cells and tissues, and when ERS pathways were blocked, the inhibiting effect was even more pronounced. Therefore, the recombinant Newcastle disease virus rL-hIFN-λ1-induced apoptosis of A549 cells is connected to ER stress and could be a promising therapeutic agent for lung adenocarcinoma.

Long non-coding RNA H19 promotes glucose metabolism and cell growth in malignant melanoma via miR-106a-5p/E2F3 axis.

PURPOSE: lncRNA H19 has been considered as an oncogenic lncRNA in many human tumours. In the present study, we identify the role and molecular mechanism of lncRNA H19 in melanoma. METHOD: QRT-PCR was used to detect the expression of lncRNA H19 and E2F3 was detected in melanoma tissues. Cell counting kit-8 (CCK8), representative metabolites analysis was used to explore the biological function of lncRNA H19, miR-106a-5p and E2F3 in melanoma cells. Bioinformatics, luciferase reporter assays, MS2-RIP and RNA pull-down assay was used to demonstrate the molecular mechanism of lncRNA H19 in melanoma. We further test the function of lncRNA H19 in vivo though Xenograft tumour assay. RESULTS: We found that lncRNA H19 was increased in melanoma tissue, and lncRNA H19 was correlated with poor prognosis of melanoma patients. miR-106a-5p acts as a tumour suppressor in melanoma by targeting E2F3. E2F3 affects the melanoma cell glucose metabolism and growth. We also demonstrated that lncRNA H19 may function as the sponge of miR-106a-5p to up-regulate E2F3 expression, and consequently promote the glucose metabolism and growth of melanoma. CONCLUSIONS: This result elucidates a new mechanism for lncRNA H19 in melanoma development and provides a survival indicator and potential therapeutic target for melanoma patients.

Recombinant Newcastle disease virus rL-RVG enhances the apoptosis and inhibits the migration of A549 lung adenocarcinoma cells via regulating alpha 7 nicotinic acetylcholine receptors in vitro.

BACKGROUND: The aim of this study were to investigate the possible pro-apoptotic mechanisms of the recombinant Newcastle disease virus (NDV) strain rL-RVG, which expresses the rabies virus glycoprotein, in A549 lung adenocarcinoma cells via the regulation of alpha 7 nicotinic acetylcholine receptors (α7 nAChRs) and to analyze the relationships between α7 nAChR expression in lung cancer and the clinical pathological features. METHODS: α7 nAChR expression in A549, LΑ795, and small-cell lung carcinoma (SCLC) cells, among others, was detected using reverse transcription polymerase chain reaction (RT-PCR). The optimal α7 nAChR antagonist and agonist concentrations for affecting A549 lung adenocarcinoma cells were detected using MTT assays. The α7 nAChR expression in A549 cells after various treatments was assessed by Western blot, immunofluorescence and RT-PCR analyses. Apoptosis in the various groups was also monitored by Western blot and TUNEL assays, followed by the detection of cell migration via transwell and scratch tests. Furthermore, α7 nAChR expression was examined by immunohistochemistry in lung cancer tissue samples from 130 patients and 40 pericancerous tissue samples, and the apoptotis in lung adenocarcinoma tissue was detected by Tunel assay, Then, the expression levels and clinicopathological characteristics were analyzed. RESULTS: Of the A549, LΑ795, SCLC and U251 cell lines, the A549 cells exhibited the highest α7 nAChR expression. The cells infected with rL-RVG exhibited high RVG gene and protein expression. The rL-RVG group exhibited weaker α7 nAChR expression compared with the methyllycaconitine citrate hydrate (MLA, an α7 nAChR antagonist) and NDV groups. At the same time, the MLA and rL-RVG treatments significantly inhibited proliferation and migration and promoted apoptosis in the lung cancer cells (P < 0.05). The expression of α7 nAChR was upregulated in lung cancer tissue compared with pericancerous tissue (P = 0.000) and was significantly related to smoking, clinical tumor-node-metastases stage, and histological differentiation (P < 0.05). The AI in lung adenocarcinoma tissue in high-medium differentiation group was lower than that in low differentiation group (p < 0.01). CONCLUSIONS: An antagonist of α7 nAChR may be used as a molecular target for lung adenocarcinoma therapy. Recombinant NDV rL-RVG enhances the apoptosis and inhibits the migration of A549 lung adenocarcinoma cells by regulating α7 nAChR signaling pathways.

Polygonatum odoratum lectin promotes BECN1 expression and induces autophagy in malignant melanoma by regulation of miR1290.

Autophagy is not only a survival response to growth-factor or nutrient deprivation but also an important mechanism for tumor-cell suicide, including melanoma. Polygonatum odoratum lectin (POL) displays apoptosis- and autophagy-inducing effects in many human tumors. POL also inhibits the growth of melanoma cells, but its role and molecular mechanism in malignant melanoma remain unclear. In this study, we found that POL suppressed proliferation and induced autophagy in melanoma cells. miR1290 was upregulated and inhibited autophagy in melanoma. BECN1 is the direct functional effector of miR1290. Furthermore, we found that POL promoted BECN1 expression though inhibition of miR1290, thus inducing melanoma-cell autophagy. This finding elucidates a new role and mechanism for POL in melanoma, and provides a potential antineoplastic agent for melanoma treatment.

Enhanced circulating ILC2s and MDSCs may contribute to ensure maintenance of Th2 predominant in patients with lung cancer.

Group 2 innate lymphoid cells (ILC2s) were demonstrated to be involved in the initiation and coordination of type 2 T helper cell (Th2) responses. Myeloid­derived suppressor cells (MDSCs) have received a great deal of attention for their role in creating an immunosuppressive microenvironment in cancer­bearing hosts. However, the contributions of ILC2s in the occurrence and development of lung cancer, and the association between ILC2s and Th2 or MDSCs in lung cancer remain to be elucidated. In the present study, 36 patients newly diagnosed with lung cancer based on the guidelines of the International Union Against Cancer Tumor Node Metastasis were included. The frequencies of ILC2s and MDSCs in peripheral blood mononuclear cells were determined, and the mRNA expression levels of ILC2s or Th2­related transcription factors and cytokines, and MDSCs­related products were assessed. The results demonstrated that the frequencies of the circulatory ILC2s and MDSCs were enhanced in lung cancer patients, as were ILC2­related transcription factors and cytokines in peripheral blood. A positive correlation was identified between the Th2­dominated phenotype and the expression levels of ILC2s­associated cytokines or transcription factors. In addition, increased autophagy related 1 was closely associated with Th2­associated transcription factors. It was demonstrated that ILC2s and MDSCs were clearly upregulated and accompanied by a predominant Th2 phenotype in patients with lung cancer; this may lead to new immunotherapy approaches for lung cancer based on the associated metabolites and cytokines.