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MicroRNAs in seminal plasma are able to discern infertile men at increased risk of developing testicular cancer

MOLECULAR ONCOLOGY 2025年第4期19卷 1188-1202页

作者： Ferrara, Carmen Battaglia, Rosalia caponnetto, angela Fazzio, Anna Stella, Michele Barbagallo, Cristina Musso, Nicolo Lunelio, Federica Vento, Maria Elena Borzi, Placido Scollo, Paolo Barbagallo, Davide Ragusa, Marco Pernagallo, Salvatore Di Pietro, Cinzia Univ Catania Dept Biomed & Biotechnol Sci Sect Biol & Genet G Sichel I-95123 Catania Italy Univ Enna Kore Dept Med & Surg Enna Italy Cannizzaro Hosp IVF Unit Catania Italy Kore Univ Enna Cannizzaro Hosp Catania Maternal & Child Dept Obstet & Gynecol Div Enna Italy DESINA Genomica S L Granada Spain

Male infertility is a risk factor for the development of testicular germ cell tumors. In this study, we investigated microRNA profiles in seminal plasma to identify potential noninvasive biomarkers able to discriminate the men at highest risk of developing cancer among the infertile population. We compared the microRNA profiles of individuals affected by testicular germ cell tumors and healthy individuals with normal or impaired spermiograms using high-throughput technology and confirmed the results by single-assay digital PCR. We found that miR-221-3p and miR-222-3p were downregulated and miR-126-3p was upregulated in cancer patients compared to both infertile and fertile men. ROC curve analysis confirmed that miR-126 upregulation is able to identify cancer patients among the infertile male population. In addition, in-depth bioinformatics analysis based on weighted gene co-expression networks showed that the identified miRNAs regulate cellular pathways involved in cancer.

关键词： circulating microRNA liquid biopsy male infertility testicular germ cell tumor

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Up-regulated microRNAs in blastocoel fluid of human implanted embryos could control circuits of pluripotency and be related to embryo competence

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JOURNAL OF ASSISTED REPRODUCTION AND GENETICS 2025年 1-15页

作者： Battaglia, Rosalia caponnetto, angela Ferrara, Carmen Fazzio, Anna Barbagallo, Cristina Stella, Michele Barbagallo, Davide Ragusa, Marco Vento, Maria Elena Borzi, Placido Scollo, Paolo Carli, Luca Feichtinger, Michael Kasapi, Evangelia Tsakos, Elias Palini, Simone Sierka, Wojciech Pecorino, Basilio Campitiello, Maria Rosaria Ronsini, Carlo Purrello, Michele Valerio, Domenico Longobardi, Salvatore D'Hooghe, Thomas Di Pietro, Cinzia Univ Catania Dept Biomed & Biotechnol Sci Sect Biol & Genet G Sichel I-95123 Catania Italy Univ Enna Kore Dept Med & Surg I-94100 Enna Italy Cannizzaro Hosp IVF Unit Catania Italy Wunschbaby Inst Feichtinger Vienna Austria EmbryoClin IVF Thessaloniki Greece Cervesi Hosp IVF Unit Cattolica Italy Gyncentrum Sp Zelazna 1 PL-40851 Katowice Poland ASL Salerno Dept Obstet & Gynecol & Physiopathol Human Reprod Salerno Italy Univ Campania Luigi Vanvitelli Dept Woman Child & Gen & Specialized Surg Naples Italy Inst Genet Res IRG I-80143 Naples Italy Merck Serono SpA Global Clin Dev Rome Italy Merck KGaA Darmstadt Germany KU Leuven Univ Leuven Dept Dev & Regenerat Biomed Sci Grp Leuven Belgium

Purpose The paper aims to investigate the biological role of microRNAs secreted by preimplantation embryo into the blastocoel fluid and to detect a distinctive molecular signature for identifying embryos with the highest implantation potential. Methods We carried on a multicenter retrospective study involving five European IVF centers. We collected 112 blastocoel fluid samples from embryos on day 5 post-fertilization, cultured individually, along with data on blastocyst grade and embryo transfer outcomes. Using a custom TLDA Array, we compared the expression levels of 89 miRNAs between 33 fluids from high-quality implanted embryos and 30 fluids from high-quality not-implanted embryos. Expression differences were assessed using SAM and t-test. Additionally, correlation and function enrichment analysis and network construction were conducted to identify the biological roles of deregulated microRNAs. Results We identified six up-regulated microRNAs in the blastocoel fluid from implanted embryos, significantly and positively correlated across all samples (r >= 0.7;P <= 0.05). They could take part in pluripotency circuits, regulating and being regulated by transcription factors associated with stemness, cell growth, and embryo development. The ROC curve analysis confirmed the potential of these miRNAs as implantation classifiers. Conclusion The six miRNAs up-regulated in blastocoel fluid from implanted embryos may represent a functional molecular signature for evaluating blastocyst quality and identifying the most competent embryos. Their evaluation associated with non-invasive preimplantation genetic testing, integrating epigenetic and genomic analyses, could enhance implantation grade and allow for identification of the euploid embryo not able to implant.

关键词： Blastocoel fluid Embryo development Embryo implantation Human blastocyst MicroRNAs

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Molecular Crosstalking among Noncoding RNAs: A New Network Layer of Genome Regulation in Cancer

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INTERNATIONAL JOURNAL OF GENOMICS 2017年第1期2017卷 1-17页

作者： Ragusa, Marco Barbagallo, Cristina Brex, Duilia caponnetto, angela Cirnigliaro, Matilde Battaglia, Rosalia Barbagallo, Davide Di Pietro, Cinzia Purrello, Michele Univ Catania Dept BioMed Sci & Biotechnol Sect Biol & Genet G Sichel BioMol Genome & Complex Syst BioMed Unit BMGS Uni Catania Italy IRCCS Associaz Oasi Maria SS Inst Res Mental Retardat & Brain Aging Troina Enna Italy

Over the past few years, noncoding RNAs (ncRNAs) have been extensively studied because of the significant biological roles that they play in regulation of cellular mechanisms. ncRNAs are associated to higher eukaryotes complexity;accordingly, their dysfunction results in pathological phenotypes, including cancer. To date, most research efforts have been mainly focused on how ncRNAs could modulate the expression of protein-coding genes in pathological phenotypes. However, recent evidence has shown the existence of an unexpected interplay among ncRNAs that strongly influences cancer development and progression. ncRNAs can interact with and regulate each other through various molecular mechanisms generating a complex network including different species of RNAs (e.g., mRNAs, miRNAs, lncRNAs, and circRNAs). Such a hidden network of RNA-RNA competitive interactions pervades and modulates the physiological functioning of canonical protein-coding pathways involved in proliferation, differentiation, and metastasis in cancer. Moreover, the pivotal role of ncRNAs as keystones of network structural integrity makes them very attractive and promising targets for innovative RNA-based therapeutics. In this review we will discuss: (1) the current knowledge on complex crosstalk among ncRNAs, with a special focus on cancer;and (2) the main issues and criticisms concerning ncRNAs targeting in therapeutics.

关键词： NON-coding RNA CROSS-talk (Biology) GENE expression PROTEIN expression GENETIC regulation MOLECULAR interactions CELL proliferation CELL differentiation

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Molecular profiling of follicular fluid microRNAs in young women affected by Hodgkin lymphoma

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REPRODUCTIVE BIOMEDICINE ONLINE 2021年第6期43卷 1045-1056页

作者： caponnetto, angela Battaglia, Rosalia Ragusa, Marco Barbagallo, Davide Lunelio, Federica Borzi, Placido Scollo, Paolo Purrello, Michele Vento, Maria Elena Di Pietro, Cinzia Univ Catania Dept Biomed & Biotechnol Sci Sect Biol & Genet Giovanni Sichel I-95123 Catania Italy Oasi Res Inst IRCCS I-94018 Troina Italy Univ Catania Interdisciplinary Res Ctr Diag & Therapy Brain Tu I-95123 Catania Italy Cannizzaro Hosp IVF Unit Catania Italy

Research question: Treatments for Hodgkin lymphoma have improved but one of their common effects is gonadal toxicity, which contributes to fertility damage of patients and induces temporary or irreversible loss of fertility. Could micro-RNA (miRNA) expression profiles in follicular fluid be influenced by Hodgkin lymphoma? Could their alteration affect molecular pathways involved in follicle growth and oocyte maturation? Design: miRNA expression profile was investigated in follicular fluid samples from young women affected by Hodgkin lymphoma compared with healthy controls by NanoString technology. Bioinformatic analysis was used to verify miRNA involvement in follicle development and miRNA deregulation with Hodgkin lymphoma in a larger cohort of follicular fluid samples was confirmed by real-time quantitative polymerase chain reaction. Results: Thirteen miRNAs are deregulated in Hodgkin lymphoma samples compared with controls and are involved in molecular pathways related to cancer, gametogenesis and embryogenesis. Among them, let-7b-5p, miR-423-5p, miR-503-5p, miR-574-5p and miR-1303 are implicated in biological processes related to follicle development and oocyte maturation. Let-7b-5p holds the central position in the regulatory network of miRNA-mRNA interactions, has the highest number of mRNA target genes shared with the other differentially expressed miRNAs and is significantly downregulated in Hodgkin lymphoma follicular fluid samples. Conclusions: These data led us to question the potential influence of miRNA deregulation on oocyte quality. Further studies are needed to verify the reproductive potential of young patients with Hodgkin lymphoma before starting chemotherapy protocols and an adequate protocol of fertility preservation needs to be guaranteed.

关键词： Follicular fluid Hodgkin lymphoma Let-7b-5p miRNA profiling

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Down-regulation of long non-coding RNAs in reproductive aging and analysis of the lncRNA-miRNA-mRNA networks in human cumulus cells

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JOURNAL OF ASSISTED REPRODUCTION AND GENETICS 2022年第4期39卷 919-931页

作者： caponnetto, angela Battaglia, Rosalia Ferrara, Carmen Vento, Maria Elena Borzi, Placido Paradiso, Marianna Scollo, Paolo Purrello, Michele Longobardi, Salvatore D'Hooghe, Thomas Valerio, Domenico Di Pietro, Cinzia Univ Catania Dept Biomed & Biotechnol Sci Sect Biol & Genet Giovanni Sichel I-95123 Catania Italy Cannizzaro Hosp IVF Unit Catania Italy Merck Serono SpA Global Clin Dev Rome Italy Healthcare Business Merck KGaA R&D Healthcare Global Med Affairs Fertil Frankfurter Str 250 D-64293 Darmstadt Germany Inst Genet Res IRG I-80143 Naples Italy

Purpose Long non-coding RNAs (lncRNAs) control gene expression at multiple levels. By interacting with microRNAs (miRNAs), they regulate their mRNA targets creating dynamic regulatory networks involved in different cellular processes. Their role in follicle development and oocyte maturation has recently emerged. lncRNA deregulation has been found associated with different pathological conditions. In this study, we identified differentially expressed lncRNAs in cumulus cells (CCs) isolated from MII oocytes of advanced maternal age women and proposed ceRNA-networks involved in signaling pathways crucial in ovarian folliculogenesis and female germ cell maturation. Methods We performed a high-throughput analysis of the expression profile of 68 lncRNAs from CCs of aged and young women by using NanoString technology. By miRNet, TarPmiR, miRTarBase, OKdb, and KEGG we predicted some ceRNA-networks involving the differentially expressed (DE) lncRNAs, miRNA interactors, and their mRNA target genes. Results We identified 28 lncRNAs down-regulated in CC samples from aged women. The analysis revealed that the miRNAs binding 11 of the DE lncRNAs and their mRNA targets are included in ceRNA-networks involved in the regulation of the PI3K-Akt, FOXO, and p53 signaling pathways. Conclusion We proposed that the lncRNA down-regulation in CCs from aged women could influence the expression of genes encoding proteins deregulated in reproductive aging. A better understanding of the interplay of lncRNA-miRNA-mRNA networks in human CCs could increase our knowledge about the mechanisms of regulation of gene expression involved in aging, lead to the development of novel therapeutics, and improve reproductive outcomes in aged women.

关键词： Regulation of gene expression lncRNAs Cumulus cells Reproductive aging ceRNA-networks

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LncRNA UCA1, Upregulated in CRC Biopsies and Downregulated in Serum Exosomes, Controls mRNA Expression by RNA-RNA Interactions

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MOLECULAR THERAPY NUCLEIC ACIDS 2018年 12卷 229-241页

作者： Barbagallo, Cristina Brex, Duilia caponnetto, angela Cimigliaro, Matilde Scalia, Marina Magnano, Antonio Caltabiano, Rosario Barbagallo, Davide Biondi, Antonio Cappellani, Alessandro Basile, Francesco Di Pietro, Cinzia Purrello, Michele Ragusa, Marco Univ Catania Sect Biol & Genet G Sichel Dept Biomed & Biotechnol Sci Via Santa Sofia 87 I-95123 Catania Italy Vittorio Emanuele Hosp Digest Endoscopy Serv I-95124 Catania Italy Univ Catania Dept Med & Surg Sci & Adv Technol GF Ingrassia I-95123 Catania Italy Univ Catania Vittorio Emanuele Hosp Dept Surg I-95124 Catania Italy IRCCS Assoc Oasi Maria SS Inst Res Mental Retardat & Brain Aging I-94018 Troina EN Italy

Long non-coding RNAs (lncRNAs) and circular RNAs (circRNAs) contribute to the onset of many neoplasias through RNA-RNA competitive interactions;in addition, they could be secreted by cancer cells into biological fluids, suggesting their potential diagnostic application. By analyzing the expression of 17 lncRNAs and 31 circRNAs in biopsies and serum exosomes from colorectal cancer (CRC) patients through qRT-PCR, we detected CCAT1, CCAT2, HOTAIR, and UCA1 upregulation and CDR1AS, MALAT1, and TUG1 downregulation in biopsies. In serum exosomes, UCA1 was downregulated, while circHIPK3 and TUG1 were upregulated. Combined receiver operating characteristic (ROC) curves of TUG1:UCA1 and circHIPK3:UCA1 showed high values of sensitivity and specificity. Through in vitro (i.e., RNA silencing and mitogen-activated protein kinase [MAPK] inhibition) and in silico analyses (i.e., expression correlation and RNA-RNA-binding prediction), we found that UCA1 could (1) be controlled by MAPKs through CEBPB;(2) sequester miR-135a, miR-143, miR-214, and miR-1271, protecting ANLN, BIRC5, IPO7, KIF2A, and KIF23 from microRNA (miRNA)-induced degradation;and (3) interact with mRNA 3'-UTRs, preventing miRNA binding. UCA1 and its co-regulated antisense LINC01764 could interact and reciprocally mask their own miRNA-binding sites. Functional enrichment analysis of the RNA-RNA network controlled by UCA1 suggested its potential involvement in cellular migration. The UCA1 regulatory axis would represent a promising target to develop innovative RNA-based therapeutics against CRC.

关键词： colorectal cancer ceRNA network circRNAs lncRNAs miRNAs LINC01764 TUG1 circHIPK3 CEBPB

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Internalization of Pegylated Er:Y2O3 Nanoparticles inside HCT-116 Cancer Cells: Implications for Imaging and Drug Delivery

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ACS APPLIED NANO MATERIALS 2023年第20期6卷 19126-19135页

作者： Chiechio, Regina Maria caponnetto, angela Battaglia, Rosalia Ferrara, Carmen Butera, Ester Musumeci, Paolo Reitano, Riccardo Ruffino, Francesco Maccarrone, Giuseppe Di Pietro, Cinzia Marchi, Valerie Lanzano, Luca Arena, Giovanni Grasso, Alfina Copat, Chiara Ferrante, Margherita Contino, Annalinda Univ Catania Dipartimento Sci Biomed & Biotecnol Sez Biol & Genet G Sichel I-95123 Catania Italy Univ Catania Dipartimento Sci Chim I-95125 Catania Italy Univ Rennes 1 Inst Sci Chim Rennes CNRS UMR 6226 F-35042 Rennes France Univ Catania Dipartimento Fis & Astron Ettore Majorana I-95123 Catania Italy CNR Ist Microelettron & i Microsistemi CNR IMM I-95123 Catania Italy Univ Catania Dept Med Surg Sci & Adv Technol GF Ingrassia Environm & Food Hyg Labs LIAA I-95124 Catania Italy

Lanthanide-doped nanoparticles, featuring sharp emission peaks with narrow bandwidth, exhibit high downconversion luminescence intensity, making them highly valuable in the fields of bioimaging and drug delivery. High-crystallinity Y2O3 nanoparticles (NPs) doped with Er3+ ions were functionalized by using a pegylation procedure to confer water solubility and biocompatibility. The NPs were thoroughly characterized using transmission electron microscopy (TEM), inductively coupled plasma mass spectrometry (ICP-MS), and photoluminescence measurements. The pegylated nanoparticles were studied both from a toxicological perspective and to demonstrate their internalization within HCT-116 cancer cells. Cell viability tests allowed for the identification of the "optimal" concentration, which yields a detectable fluorescence signal without being toxic to the cells. The internalization process was investigated using a combined approach involving confocal microscopy and ICP-MS. The obtained data clearly indicate the efficient internalization of NPs into the cells with emission intensity showing a strong correlation with the concentrations of nanoparticles delivered to the cells. Overall, this research contributes significantly to the fields of nanotechnology and biomedical research, with noteworthy implications for imaging and drug delivery applications.

关键词： Nanoparticles luminescence lanthanides pegylation cancer cells internalization

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Competing endogenous RNA network mediated by circ_3205 in SARS-CoV-2 infected cells

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CELLULAR AND MOLECULAR LIFE SCIENCES 2022年第2期79卷 75-75页

作者： Barbagallo, Davide Palermo, Concetta Ilenia Barbagallo, Cristina Battaglia, Rosalia caponnetto, angela Spina, Vittoria Ragusa, Marco Di Pietro, Cinzia Scalia, Guido Purrello, Michele Univ Catania Dept Biomed & Biotechnol Sci Sect Biol & Genet Giovanni Sichel I-95123 Catania Italy AOU Policlin Vittorio Emanuele UOC Lab Anal Unit I-95123 Catania Italy Univ Catania Dept Biomed & Biotechnol Sci Sect Med Biochem I-95123 Catania Italy Univ Catania Dept Biomed & Biotechnol Sci Sect Microbiol I-95123 Catania Italy

Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) is a new member of the Betacoronaviridae family, responsible for the recent pandemic outbreak of COVID-19. To start exploring the molecular events that follow host cell infection, we queried VirusCircBase and identified a circular RNA (circRNA) predicted to be synthesized by SARS-CoV-2, circ_3205, which we used to probe: (i) a training cohort comprised of two pools of cells from three nasopharyngeal swabs of SARS-CoV-2 infected (positive) or uninfected (negative, UCs) individuals;(ii) a validation cohort made up of 12 positive and 3 negative samples. The expression of circRNAs, miRNAs and miRNA targets was assayed through real-time PCR. CircRNA-miRNA interactions were predicted by TarpMiR, Analysis of Common Targets for circular RNAs (ACT), and STarMir tools. Enrichment of the biological processes and the list of predicted miRNA targets were retrieved from DIANA miRPath v3.0. Our results showed that the predicted SARS-CoV-2 circ_3205 was expressed only in positive samples and its amount positively correlated with that of SARS-CoV-2 Spike (S) mRNA and the viral load (r values = 0.80952 and 0.84867, Spearman's correlation test, respectively). Human (hsa) miR-298 was predicted to interact with circ_3205 by all three predictive tools. KCNMB4 and PRKCE were predicted as hsa-miR-298 targets. Interestingly, the function of both is correlated with blood coagulation and immune response. KCNMB4 and PRKCE mRNAs were upregulated in positive samples as compared to UCs (6 and 8.1-fold, p values = 0.049 and 0.02, Student's t test, respectively) and their expression positively correlated with that of circ_3205 (r values = 0.6 and 0.25, Spearman's correlation test, respectively). We propose that our results convincingly suggest that circ_3205 is a circRNA synthesized by SARS-CoV-2 upon host cell infection and that it may behave as a competitive endogenous RNA (ceRNA), sponging hsa-miR-298 and contributing to the upregulati

关键词： Coronavirus disease 2019 circRNA miRNA sponge microRNA Gene expression regulatory network

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Er:Y2O3 and Nd:Y2O3 Nanoparticles: Synthesis, Pegylation, Characterization and Study of Their Luminescence Properties

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CHEMOSENSORS 2023年第1期11卷 20页

作者： Chiechio, Regina Maria Battaglia, Rosalia caponnetto, angela Butera, Ester Franzo, Giorgia Reitano, Riccardo Purrello, Michele Ragusa, Marco Barbagallo, Davide Barbagallo, Cristina Di Pietro, Cinzia Marchi, Valerie Lo Faro, Maria Jose Contino, Annalinda Maccarrone, Giuseppe Musumeci, Paolo Univ Catania Dipartimento Fis & Astron Ettore Majorana Via Santa Sofia 64 I-95123 Catania Italy Univ Rennes 1 Inst Sci Chim Rennes CNRS UMR 6226 Ave Gen Leclerc F-35042 Rennes France Consiglio Nazl Ric CNR Ist Microelettron & i Microsistemi IMM Via S Sofia 64 I-95123 Catania Italy Univ Catania Dipartimento Sci Biomed & Biotecnolog Sez Biol & Genet G Sichel Via S Sofia 89 I-95123 Catania Italy Univ Catania Dipartimento Sci Chim Viale Andrea Doria 6 I-95125 Catania Italy

Lanthanide-doped yttrium oxide nanoparticles can display selective upconversion properties, rendering them invaluable in the field of nanomedicine for both sensing and diagnostics. Different syntheses of Er:Y2O3 and Nd:Y2O3 nanoparticles (NPs) were studied and optimized to obtain small particles of regular shape and good crystallinity. The morphological and compositional characterizations of the nanoparticles were obtained with different techniques and showed that both Er:Y2O3 and Nd:Y2O3 NPs were well dispersed, with dimensions of the order of a few tens of nanometers. The photoluminescence and cathodoluminescence measurements showed that both Er:Y2O3 and Nd:Y2O3 NPs had good emission as well as upconversion. The nanophosphors were functionalized by a pegylation procedure to suppress unwanted reactions of the NPs with other biological components, making the NP systems biocompatible and the NPs soluble in water and well dispersed. The pegylated core/shell nanoparticles showed the same morphological and optical characteristics as the core, promoting their strategic role as photoactive material for theragnostics and biosensing.

关键词： nanoparticles upconversion luminescence lanthanides pegylation cathodoluminescence

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Smoking Cessation and Reduction in Schizophrenia (SCARIS) with e-cigarette: study protocol for a randomized control trial

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TRIALS 2014年第1期15卷 1-8页

作者： caponnetto, Pasquale Polosa, Riccardo Auditore, Roberta Minutolo, Giuseppe Signorelli, Maria Maglia, Marilena Alamo, angela Palermo, Filippo Aguglia, Eugenio Univ Catania Dept Clin & Mol Biomed Policlin V Emanuele Smoking Prevent Cessat CtrAOU Catania Italy Univ Catania Dept Clin & Mol Biomed AziendaOsped Univ Policlin V Emanuele Inst Internal Med Catania Italy CTA Villa Chiara Psychiat Rehabil Clin & Res I-95030 Catania Italy Univ Catania Policlin Vittorio Emanuele Hosp Dept Clin & Mol Biomed UOPI Psychiat I-95100 Catania Italy Univ Catania AO Garibaldi Nesima Dept Clin & Mol Biomed Catania Italy

Background: It is well established in studies across several countries that tobacco smoking is more prevalent among schizophrenic patients than the general population. Electronic cigarettes are becoming increasingly popular with smokers worldwide. To date there are no large randomized trials of electronic cigarettes in schizophrenic smokers. A well-designed trial is needed to compare efficacy and safety of these products in this special population. Methods/Design: Intervention: We have designed a randomized controlled trial investigating the efficacy and safety of electronic cigarette. The trial will take the form of a prospective 12-month randomized clinical study to evaluate smoking reduction, smoking abstinence and adverse events in schizophrenic smokers not intending to quit. We will also monitor quality of life, neurocognitive functioning and measure participants' perception and satisfaction of the product. Outcome measures: A >= 50% reduction in the number of cigarettes/day from baseline, will be calculated at each study visit ("reducers"). Abstinence from smoking will be calculated at each study visit ("quitters"). Smokers who leave the study protocol before its completion and will carry out the Early Termination Visit or who will not satisfy the criteria of "reducers" and "quitters" will be defined "non responders". Statistical analysis: The differences of continuous variables between the three groups will be evaluated with the Kruskal-Wallis Test, followed by the Dunn multiple comparison test. The differences between the three groups for normally distributed data will be evaluated with ANOVA test one way, followed by the Newman-Keuls multiple comparison test. The normality of the distribution will be evaluated with the Kolmogorov-Smirnov test. Any correlations between the variables under evaluation will be assessed by Spearman r correlation. To compare qualitative data will be used the Chi-square test. Discussion: The main strengths of the SCARIS study are

关键词： Smoking cessation Smoking reduction Electronic cigarette Schizophrenia clinical trial Cognitive Addiction Quality of life

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