Synergistic Effect of an Antisense Oligonucleotide and Small Molecule on Splicing Correction of the Spinal Muscular Atrophy Gene.

Spinal muscular atrophy (SMA) is treated by increasing the level of Survival Motor Neuron (SMN) protein through correction of SMN2 exon 7 skipping or exogenous expression of SMN through gene therapy. Currently available therapies have multiple shortcomings, including poor body-wide distribution, invasive delivery, and potential negative consequences due to high doses needed for clinical efficacy. Here we test the effects of a combination treatment of a splice-correcting antisense oligonucleotide (ASO) Anti-N1 with the small compounds risdiplam and branaplam. We show that a low-dose treatment of Anti-N1 with either compound produces a synergistic effect on the inclusion of SMN2 exon 7 in SMA patient fibroblasts. Using RNA-Seq, we characterize the transcriptomes of cells treated with each compound as well as in combination. Although high doses of each individual treatment trigger widespread perturbations of the transcriptome, combination treatment of Anti-N1 with risdiplam and branaplam results in minimal disruption of gene expression. For individual genes targeted by the 3 compounds, we observe little to no additive effects of combination treatment. Overall, we conclude that the combination treatment of a splice-correcting ASO with small compounds represents a promising strategy for achieving a high level of SMN expression while minimizing the risk of off-target effects.

Structural Context of a Critical Exon of Spinal Muscular Atrophy Gene.

Humans contain two nearly identical copies of Survival Motor Neuron genes, SMN1 and SMN2. Deletion or mutation of SMN1 causes spinal muscular atrophy (SMA), one of the leading genetic diseases associated with infant mortality. SMN2 is unable to compensate for the loss of SMN1 due to predominant exon 7 skipping, leading to the production of a truncated protein. Antisense oligonucleotide and small molecule-based strategies aimed at the restoration of SMN2 exon 7 inclusion are approved therapies of SMA. Many cis-elements and transacting factors have been implicated in regulation of SMN exon 7 splicing. Also, several structural elements, including those formed by a long-distance interaction, have been implicated in the modulation of SMN exon 7 splicing. Several of these structures have been confirmed by enzymatic and chemical structure-probing methods. Additional structures formed by inter-intronic interactions have been predicted by computational algorithms. SMN genes generate a vast repertoire of circular RNAs through inter-intronic secondary structures formed by inverted Alu repeats present in large number in SMN genes. Here, we review the structural context of the exonic and intronic cis-elements that promote or prevent exon 7 recognition. We discuss how structural rearrangements triggered by single nucleotide substitutions could bring drastic changes in SMN2 exon 7 splicing. We also propose potential mechanisms by which inter-intronic structures might impact the splicing outcomes.

High Concentration of an ISS-N1-Targeting Antisense Oligonucleotide Causes Massive Perturbation of the Transcriptome.

Intronic splicing silencer N1 (ISS-N1) located within Survival Motor Neuron 2 (SMN2) intron 7 is the target of a therapeutic antisense oligonucleotide (ASO), nusinersen (Spinraza), which is currently being used for the treatment of spinal muscular atrophy (SMA), a leading genetic disease associated with infant mortality. The discovery of ISS-N1 as a promising therapeutic target was enabled in part by Anti-N1, a 20-mer ASO that restored SMN2 exon 7 inclusion by annealing to ISS-N1. Here, we analyzed the transcriptome of SMA patient cells treated with 100 nM of Anti-N1 for 30 h. Such concentrations are routinely used to demonstrate the efficacy of an ASO. While 100 nM of Anti-N1 substantially stimulated SMN2 exon 7 inclusion, it also caused massive perturbations in the transcriptome and triggered widespread aberrant splicing, affecting expression of essential genes associated with multiple cellular processes such as transcription, splicing, translation, cell signaling, cell cycle, macromolecular trafficking, cytoskeletal dynamics, and innate immunity. We validated our findings with quantitative and semiquantitative PCR of 39 candidate genes associated with diverse pathways. We also showed a substantial reduction in off-target effects with shorter ISS-N1-targeting ASOs. Our findings are significant for implementing better ASO design and dosing regimens of ASO-based drugs.

Spinal muscular atrophy: Broad disease spectrum and sex-specific phenotypes.

Spinal muscular atrophy (SMA) is one of the major genetic disorders associated with infant mortality. More than 90% of cases of SMA result from deletions of or mutations in the Survival Motor Neuron 1 (SMN1) gene. SMN2, a nearly identical copy of SMN1, does not compensate for the loss of SMN1 due to predominant skipping of exon 7. The spectrum of SMA is broad, ranging from prenatal death to infant mortality to survival into adulthood. All tissues, including brain, spinal cord, bone, skeletal muscle, heart, lung, liver, pancreas, gastrointestinal tract, kidney, spleen, ovary and testis, are directly and/or indirectly affected in SMA. Accumulating evidence on impaired mitochondrial biogenesis and defects in X chromosome-linked modifying factors, coupled with the sexual dimorphic nature of many tissues, point to sex-specific vulnerabilities in SMA. Here we review the role of sex in the pathogenesis of SMA.

The First Orally Deliverable Small Molecule for the Treatment of Spinal Muscular Atrophy.

Spinal muscular atrophy (SMA) is 1 of the leading causes of infant mortality. SMA is mostly caused by low levels of Survival Motor Neuron (SMN) protein due to deletion of or mutation in the SMN1 gene. Its nearly identical copy, SMN2, fails to compensate for the loss of SMN1 due to predominant skipping of exon 7. Correction of SMN2 exon 7 splicing by an antisense oligonucleotide (ASO), nusinersen (Spinraza™), that targets the intronic splicing silencer N1 (ISS-N1) became the first approved therapy for SMA. Restoration of SMN levels using gene therapy was the next. Very recently, an orally deliverable small molecule, risdiplam (Evrysdi™), became the third approved therapy for SMA. Here we discuss how these therapies are positioned to meet the needs of the broad phenotypic spectrum of SMA patients.

RNA in spinal muscular atrophy: therapeutic implications of targeting.

INTRODUCTION: Spinal muscular atrophy (SMA) is caused by low levels of the Survival Motor Neuron (SMN) protein due to deletions of or mutations in the SMN1 gene. Humans carry another nearly identical gene, SMN2, which mostly produces a truncated and less stable protein SMNΔ7 due to predominant skipping of exon 7. Elevation of SMN upon correction of SMN2 exon 7 splicing and gene therapy have been proven to be the effective treatment strategies for SMA. AREAS COVERED: This review summarizes existing and potential SMA therapies that are based on RNA targeting.We also discuss the mechanistic basis of RNA-targeting molecules. EXPERT OPINION: The discovery of intronic splicing silencer N1 (ISS-N1) was the first major step towards developing the currently approved antisense-oligonucleotide (ASO)-directed therapy (SpinrazaTM) based on the correction of exon 7 splicing of the endogenous SMN2pre-mRNA. Recently, gene therapy (Zolgensma) has become the second approved treatment for SMA. Small compounds (currently in clinical trials) capable of restoring SMN2 exon 7 inclusion further expand the class of the RNA targeting molecules for SMA therapy. Endogenous RNA targets, such as long non-coding RNAs, circular RNAs, microRNAs and ribonucleoproteins, could be potentially exploited for developing additional SMA therapies.

Programa Mais Médicos: contribuições aos processos de trabalho e desafios para a integralidade do cuidado na Estratégia Saúde da Família / Mais Médicos (More Doctors) Program: contributions to the work processes and challenges for a comprehensive health care in Family Health Strategy / Programa Mais Médicos (Más Médicos): contribuciones a los procesos de trabajo y desafíos para la integralidad del cuidado en la Estrategia Salud de la Familia

Rev Bras Med Fam Comunidade. Rio de Janeiro, 2020 Jan-Dez; 15(42):21491Programa Mais Médicos: contribuições aos processos de trabalho e desafios para a integralidade do cuidado na Estratégia Saúde da FamíliaPablo de Almeida Boiteux1, Thiago Dias Sarti1, Rita de Cássia Duarte Lima1Mais Médicos (More Doctors) Program: contributions to the work processes and challenges for a comprehensive health care in Family Health StrategyEste artigo analisa os efeitos da inserção de uma médica cubana do Programa Mais Médicos nos processos de trabalho de uma equipe de Saúde da Família, discutindo suas contribuições e desafios à integralidade do cuidado em saúde. Trata-se de uma pesquisa qualitativa, descritivo-exploratória, do tipo estudo de caso, realizada em município do estado do Espírito Santo, Brasil, mediante utilização de observação participante, complementada com entrevistas em profundidade e um grupo focal. A análise dos dados foi fundamentada nas teorias do processo de trabalho e da produção intersubjetiva do cuidado em saúde. Os resultados indicam contribuições na ampliação de ações assistenciais e no fortalecimento dos vínculos entre equipe e usuários. Entre os desafios, a persistência de práticas segmentadas por categorias profissionais, subordinadas à figura e aos saberes (bio)médicos, com limitada interação interprofissional e equipe-comunidade na construção de projetos comuns de cuidado.

This article analyzes the effects of the insertion of a Cuban doctor from the Mais Médicos (More Doctors) Program into the work processes of a Family Health Strategy team, discussing its contributions and challenges to the comprehensiveness of health care. This is a qualitative, descriptive-exploratory research, of the case study type, carried out in a city of the state of Espírito Santo, Brazil, using participant observation, in-depth interviews and focus group. The analysis was based on theories of the work process and the intersubjective production of health care. Results indicate contributions in the expansion of assistance actions and in the strengthening of the links between team and users. Among the challenges, the persistence of practices segmented by professional categories, subordinate to the (bio)medical figure and knowledge, with limited interprofessional and team-community interactions in the construction of common projects of care.

Este artículo analiza los efectos de la inserción de una médica cubana del Programa Mais Médicos en los procesos de trabajo de un equipo de Salud de la Familia, discutiendo sus contribuciones y desafíos a la integralidad del cuidado en salud. Es una investigación descriptiva-exploratoria, del tipo estudio de caso, realizada en municipio del estado de Espírito Santo, Brasil, mediante utilización de observación participante, complementada con entrevistas en profundidad y un grupo focal. El análisis fue fundamentado en teorías del proceso de trabajo y de la producción intersubjetiva del cuidado en salud. Los resultados indican contribuciones en la ampliación de y en el fortalecimiento de vínculos entre equipo y usuarios. Entre los desafíos, la persistencia de prácticas profesionales segmentadas, subordinadas al médico y sus saberes, con limitada interacción interprofesional y equipo-comunidad en la construcción de proyectos comunes de cuidado.

Infecção marinha: abordagem na Atenção Primária / Marine infection: approach in Primary Care / Infección marina: enfoque en la Atención Primaria

Introdução: Cidades litorâneas possuem atividades econômicas como a pesca e o turismo que promovem o contato com a água do mar e, consequentemente, a exposição a microrganismos raramente encontrados em outros contextos e muitas vezes de diagnóstico tardio, podendo resultar em morbidade ou morte significativa. Objetivo: Essa revisão da literatura objetiva mostrar a importância da suspeição da infecção marinha na Atenção Primária, sua etiologia, manifestações clínicas, tratamento, complicações e prevenção. Métodos: Foi realizada pesquisa em bases de dados eletrônicos (SciELO, Google Acadêmico, MEDLINE e PubMed). Resultados: Foram encontrados 135 artigos e vinte foram selecionados, referentes aos anos de 2003 a 2018. Observou-se escassez de estudos que avaliam a efetividade de esquemas de antibioticoterapia e sua duração necessária. Além disso, verificou-se a ausência de classificação no Código Internacional de Doenças (CID-10) e Descritores em Ciências da Saúde (DeCS), fato que prejudica a notificação e estudos epidemiológicos sobre o assunto. Conclusão: É preciso incluir a infecção marinha no diagnóstico diferencial de casos de ferimentos crônicos de difícil diagnóstico, principalmente se houver histórico de exposição a ambiente marinho. Novos estudos se fazem necessários para avaliação da terapêutica adequada. Outrossim, é fundamental conscientizar a população quanto ao risco de infecção marinha e seus métodos de prevenção.

Introduction: Coastal cities have economic activities such as fishing and tourism that promote contact with sea water and consequent exposure to microorganisms rarely found in other contexts. Too often, late diagnosis can result in significant morbidity or death. Objective: This review of the literature aims to show the importance of the differential diagnosis of marine infection in primary care, its etiology, clinical manifestations, treatment, complications and prevention. Methods: Electronic databases (SciELO, Google Scholar, MEDLINE and PubMed) were searched. Results: 135 articles were found and twenty were selected, referring to the years 2003 to 2018. A shortage of studies evaluating the effectiveness of antibiotic therapy regimens and their necessary duration was observed. In addition, the absence of the International Code of Diseases (ICD-10) and Descriptors in Health Sciences (DeCS) was observed, which is detrimental to the epidemiological notification and study on the subject. Conclusion: It is necessary to include marine infection in the differential diagnosis of cases of chronic injuries difficult to diagnose, especially if there is a history of exposure to the marine environment. Further studies are needed to evaluate the appropriate therapy. It is also essential to educate the population about the risk of marine infection and methods of prevention.

Introducción: Las ciudades costeras tienen actividades económicas como la pesca y el turismo que promueven el contacto com la agua del mar y consecuentemente la exposición a microorganismos raramente encontrados em otros contextos y muchas veces de diagnóstico tardío, pudiendo resultar em morbilidad o muerte significativa. Objetivo: Esta revisión de la literatura objetiva mostrar la importancia de la sospecha de la infección marina en la atención primaria, su etiología, manifestaciones clínicas, tratamiento, complicaciones y prevención. Método: Se realizo investigación en bases de datos electrónicos (SciELO, Google Académico, MEDLINE y PubMed). Resultados: Se encontraron 135 artículos y veinte fueron seleccionados, referentes a los años de 2003 a 2018. Se observo escasez de estudios que evalúan la efectividad de esquemas de antibioticoterapia y suduración necesaria. Además, se verifico la ausencia de clasificación en el Código Internacional de Enfermedades (CID-10) y Descriptores em Ciencias de La Salud (DeCS), hecho que perjudica la notificación y estudio epidemiológicos sobre el tema. Conclusión: Es necesario incluir la infección marina em el diagnóstico diferencial de casos de lesiones crónicas de difícil diagnóstico, principalmente si hayun historial de exposición al ambiente marino. Los nuevos estudios se hacen necesarios para evaluar la terapéutica adecuada. También es fundamental concientizar a la población encuanto al riesgo de infección marina y sus métodos de prevención.

Capacidade funcional e risco de quedas em idosos com síndrome metabólica / Functional capacity and risk of falls in the elderly with metabolic syndrome / Capacidad funcional y riesgo de caídas en ancianos con síndrome metabólico

Introdução: A síndrome metabólica (SM) é a anormalidade metabólica mais comum na população que predispõe a eventos cardiovasculares e pode estar relacionada à baixa aptidão física e baixa capacidade funcional em idosos. O objetivo deste estudo foi avaliar a interferência das alterações metabólicas na capacidade funcional e no risco de quedas em idosos. Métodos: Trata-se de um estudo transversal realizado com 126 idosos, sendo 58 com síndrome metabólica e 68 sem síndrome metabólica, segundo os critérios da International Diabetes Federation. Os índices de Katz, Tinetti, Timed Up e Go e Berg Scale foram aplicados para avaliar a capacidade funcional. A Escala de Eficácia das Quedas-Escala Internacional (FES-I-Brasil) foi utilizada para avaliar o medo de cair. Para análise estatística, foram utilizados os testes de correlação de Mann-Whitney, Qui-quadrado e Spearman. Resultados: Houve associação dos grupos MetS e non-MetS em relação com os estratos de risco do índice TUG [p=0,02; OR=0,38; IC95% (0,16-0,91)]. Entre esses dois grupos, houve diferença significativa na média dos seguintes marcadores: escala de Berg (p=0,03); TUG (p=0,03); FES-I-Brazil (p=0,02). O índice de Kats e IPAQ não apresentaram associação significativa. Conclusões: Os idosos com SM apresentaram maior risco e medo de quedas quando comparados aos pacientes sem SM. No entanto, não houve variação na capacidade de caminhar ou no equilíbrio.

Introduction: Metabolic syndrome (MetS) is the most common metabolic abnormality in the population that predisposes to cardiovascular events and may be related to poor physical fitness and low functional capacity in the elderly. The objective of this study was to evaluate the interference of metabolic changes in functional capacity and risk of falls in the elderly. Methods: A cross-sectional study was carried out with 126 elderly subjects, 58 presented MetS and 68 without MetS, according to the criteria of the International Diabetes Federation. Katz Index, Tinetti Index, Timed Up and Go test and Berg Scale were applied to evaluate the functional capacity. Falls Efficacy Scale-International scale (FES-I-Brazil) was used to evaluate the fear of falling. For statistical analysis, the Mann-Whitney, Chi-square and Spearman correlation tests were used. Results: There was an association between the MetS and non-MetS groups in relation to the risk strata of the TUG index [p=0.02; OR=0.38; 95%CI (0.16-0.91)]. Between these two groups, there was a significant difference in the mean of the following markers: Berg scale (p=0.03); TUG (p=0.03); FES-I-Brazil (p=0.02). The Kats and IPAQ index did not show any significant association. Conclusions: Elderly patients with MetS presented higher risk and fear of falls when compared to patients without MetS. However, there was no variation in walking ability or balance.

Introducción: El síndrome metabólico (MetS) es la anormalidad metabólica más común en la población que predispone a eventos cardiovasculares y puede estar relacionado con un mal estado físico y una baja capacidad funcional en los ancianos. El objetivo de este estudio fue evaluar la interferencia de los cambios metabólicos en la capacidad funcional y el riesgo de caídas en los ancianos. Método: Se realizó un estudio transversal con 126 sujetos de edad avanzada, 58 presentaron MetS y 68 sin MetS, según los criterios de la Federación Internacional de Diabetes. Se aplicaron los índices Katz, Tinetti, Timed Up and Go y Berg Scale para evaluar la capacidad funcional. La Escala de Eficacia de las Caídas - Escala internacional (FES-I-Brasil) se utilizó para evaluar el miedo a las caídas. Para el análisis estadístico, se utilizaron las pruebas de correlación de Mann-Whitney, Chi-cuadrado y Spearman. Resultados: Hubo una asociación entre los grupos MetS y no MetS en relación con los estratos de riesgo del índice TUG [p=0.02; OR=0,38; IC 95% (0.16-0.91)]. Entre estos dos grupos, hubo una diferencia significativa en la media de los siguientes marcadores: escala de Berg (p=0.03); TUG (p=0.03); FES-I-Brasil (p=0.02). El índice de Kats e IPAQ no mostró ninguna asociación significativa. Conclusiones:Los pacientes de edad avanzada con síndrome metabólico presentaron mayor riesgo y miedo a las caídas en comparación con los pacientes sin síndrome metabólico. Sin embargo, no hubo variación en la capacidad para caminar o el equilibrio.

Pointless pondering on predatory publications.

It is a pointless pondering (thinking) on predatory (meaning greedy) publications (meaning journals) while practicing publishing through freedom of expression and or the Press where applicable. It should be noted that a weak publication will vanish (disappear) itself in an open access publishing model where contents are made available for free on the WWW. The fundamental question in this context is the definition of host (congregation) and predator (intruder). The second question is the type (data and or commercial) and subsequent measure of effect of the predator on the host. Detailed discussion on this issue or any other related issue is welcomed under the freedom of the Press yet conclusion on it will be often biased and is clearly unwarranted. The parties aware of such concerns should write to the publisher (with address for communication) to take such action within such time to stand corrected. Please be informed that ISSN is unique for each publication and portals for ISSN is distributed throughout the world in each country. This is well monitored and clearly streamlined. Therefore, NO two publication titles will be identical. Awareness from authors on misleading or misinformed or misrepresented ISSN is important and such information should be petitioned to ISSN and portals for ISSN that is distributed throughout the world with state mechanisms to monitor such activities. Academia should be self-aware on these issues and have discussions on the quality and quantity of data taken to the context. Caveat Emptor is applicable to a considerable extend among the literate community as in this case. The only problem could arise because of compromised (unregistered or mirrored) ISSN number published on the WWW which is already well regulated through DNS lookup. Therefore, parties concerned about ethical issues on scientific publishing should write to concerned publishers with known address to stand corrected or to ISSN and portals for ISSN or to DNS lookup where address is not available to correct such issues through available state mechanisms. Hence, biased advisory notes from government representations, society sponsored mass campaign through news/TV media and academic miss representation based on data collected by an individual without physical address for communication is clearly unwarranted in this regard.

Expression of transcription factor FOXP4 in laryngeal squamous cell carcinoma tissues and its effects on biological characteristics of laryngeal squamous cell carcinoma TU177 cells / 中国肿瘤生物治疗杂志

@#[Abstract] Objective:To detect the expression of transcription factor FOXP4 (Forkhead box P4) in laryngeal squamous cell carcinoma (LSCC) tissues and cell lines, and to investigate its effects on the proliferation, migration, invasion, cell cycle, and apoptosis of LSCC TU177 cells in vitro as well as to explore its relationship with epithelial-mesenchymal transition (EMT) process. Methods: A total of 40 pairs of tumor tissues and adjacent tissues that resected from LSCC patients were collected from the biological specimen bank of the Forth Hospital of Hebei Medical University between 2013 and 2015. The expression of FOXP4 in LSCC tissues and corresponding adjacent tissues was detected by qPCR. qPCR and Western blotting were used to detect the FOXP4 expression level in human LSCC cell lines (AMC-HN-8, TU177, TU686, and TU212). Small interfering RNA (si-RNA) was used to knock down FOXP4 expression in TU177 cells. The effects of FOXP4 knockdown on the proliferation, migration, invasion, cell cycle and apoptosis of TU177 cells were measured by MTS assay, clone formation assay, Transwell chamber migration and invasion assay, and flow cytometry, respectively. The mRNA levels of EMT markers N-cadherin, β-catenin, Vimentin, Twist, Snail and zine finger E box binding homeobox 1 (ZEB1) after transfection of si-FOXP4 in TU177 cells were detected by qPCR. The changes of protein levels of N-cadherin, β-catenin, Vimentin and Twist after FOXP4 knockdown were measured by Western blotting. Results: The expression of FOXP4 in LSCC tissues was significantly higher than that in adjacent tissues (P<0.05), and it was related to the TNM stage of tumors and lymph node metastasis (all P<0.05). The expression of FOXP4 in LSCC cells was higher than that in the adjacent tissues (P<0.05 or P<0.01). The expression of FOXP4 in TU177 cells transfected with si-FOXP4 was significantly lower than that in the control group (P<0.01). Compared with the control group, knocking down FOXP4 could inhibit the proliferation, migration and invasion but promote the apoptosis of TU177 cells in vitro (all P<0.01), block the cell cycle at G0/G1 phase (P<0.01), and reduce cell replication in S phase (P<0.01); in addition, knocking down FOXP4 could reduce the mRNA levels of N-cadherin, β-catenin, Vimentin, Twist, Snail, ZEB1 (P<0.05 or P<0.01) and the protein levels of N-cadherin, β-catenin, Vimentin, Twist in TU177 cells. Conclusion: The high expression of FOXP4 may be related to the occurrence and development of LSCC. FOXP4 knockdown can inhibit the proliferation, migration and invasion of laryngeal cancer cells in vitro, block cell cycle at G0/G1 phase, promote apoptosis, and may participate in the EMT process.

A cluster of COVID-19 caused by a presymptomatic case in Haikou / 预防医学

Objective@#To investigate a cluster of coronavirus disease 2019(COVID-19)caused by latent infection in Haikou,so as to provide reference for the prevention and control of COVID-19 clusters. @*Methods@#An epidemiological investigation was conducted according to the COVID-19 Prevention and Control Program(Fourth Edition). The course of diagnosis and treatment,clinical characteristics and field investigation data were collected to analyze the transmission chain and the intergeneration between cases. @*Results @#Among 39 people involved,five confirmed cases and two asymptomatic infections were found,with an attack rate of 17.95%. The cases aged from 40 to 65 years and lived in the dormitories near the farm of Dongshan. The first case(named Case 1)closely contacted with a confirmed case of COVID-19 in Chengmai from January 28 to February 10,was isolated on February 13 and developed symptoms on February 16. The other six cases(Case 2 to Case 7)shared the water source with Case 1 from January 28 to February 13(within the incubation period of Case 1). They needed to open the power switch outside Case 1's room and the water valve in Case 1's washroom before the use of water. They might be infected by contacting the doorknob and the water valve contacted by Case 1,or by the aerosol formed after Case 1 used the washroom,then infected each other when living together. The onset of Case 2 to Case 7 was earlier than Case 1,and they had no travelling history in Hubei Province 14 days before and contacted no confirmed cases except Case 1. Therefore,Case 1 was the source of the cluster during his incubation period of COVID-19. @*Conclusion@#This was a cluster of COVID-19 due to latent transmission by living in the same area and touching the same objects indirectly,which indicated that COVID-19 was infectious in the incubation period.

How RNA structure dictates the usage of a critical exon of spinal muscular atrophy gene.

Role of RNA structure in pre-mRNA splicing has been implicated for several critical exons associated with genetic disorders. However, much of the structural studies linked to pre-mRNA splicing regulation are limited to terminal stem-loop structures (hairpins) sequestering splice sites. In few instances, role of long-distance interactions is implicated as the major determinant of splicing regulation. With the recent surge of reports of circular RNA (circRNAs) generated by backsplicing, role of Alu-associated RNA structures formed by long-range interactions are taking central stage. Humans contain two nearly identical copies of Survival Motor Neuron (SMN) genes, SMN1 and SMN2. Deletion or mutation of SMN1 coupled with the inability of SMN2 to compensate for the loss of SMN1 due to exon 7 skipping causes spinal muscular atrophy (SMA), one of the leading genetic diseases of children. In this review, we describe how structural elements formed by both local and long-distance interactions are being exploited to modulate SMN2 exon 7 splicing as a potential therapy for SMA. We also discuss how Alu-associated secondary structure modulates generation of a vast repertoire of SMN circRNAs. This article is part of a Special Issue entitled: RNA structure and splicing regulation edited by Francisco Baralle, Ravindra Singh and Stefan Stamm.

A novel role of U1 snRNP: Splice site selection from a distance.

Removal of introns by pre-mRNA splicing is fundamental to gene function in eukaryotes. However, understanding the mechanism by which exon-intron boundaries are defined remains a challenging endeavor. Published reports support that the recruitment of U1 snRNP at the 5'ss marked by GU dinucleotides defines the 5'ss as well as facilitates 3'ss recognition through cross-exon interactions. However, exceptions to this rule exist as U1 snRNP recruited away from the 5'ss retains the capability to define the splice site, where the cleavage takes place. Independent reports employing exon 7 of Survival Motor Neuron (SMN) genes suggest a long-distance effect of U1 snRNP on splice site selection upon U1 snRNP recruitment at target sequences with or without GU dinucleotides. These findings underscore that sequences distinct from the 5'ss may also impact exon definition if U1 snRNP is recruited to them through partial complementarity with the U1 snRNA. In this review we discuss the expanded role of U1 snRNP in splice-site selection due to U1 ability to be recruited at more sites than predicted solely based on GU dinucleotides.

[Study of clinical outcome and prognosis in pediatric core binding factor-acute myeloid leukemia].

Objective: To analyze the clinical outcome and the prognostic factor in pediatric patients with core binding factor-acute myeloid leukemia (CBF-AML). Methods: A total of 121 newly diagnosed pediatric CBF-AML patients enrolled from Aug. 2005 to Sep. 2017 were retrospectively reviewed. Cumulative incidence of relapse (CIR), event-free survival (EFS) and overall survival (OS) rates were estimated by Kaplan-Meier method and prognostic factors were evaluated by Cox regression with SPSS. Results: Of the 121 patients, 120 patients were assessed for bone marrow remission after induction chemotherapy. 100 cases (83.3%) achieved complete remission (CR) after the first course of chemotherapy. 119 cases (99.2%) achieved CR after the second course of chemotherapy. Of the 121 patients, 13 patients (10.7%) had recurrence with the median interval of recurrence as 13.8 months (3.7 to 58.8 months). 17 patients (14.0%) died. The CIR, EFS and OS at 3 years were 12.7%, 77.5% and 82.8%, respectively. The factors including age at diagnosis, sex, initial WBC count, presence of extramedullary leukemia, C-KIT expression, additional chromosomal abnormalities, and CR after the first course of chemotherapy were analyzed by multivariate regression analysis of Cox. Multivariate analysis identified that additional chromosomal abnormalities was the only independent risk factor affecting OS (HR=4.289, 95%CI 1.070-17.183, P=0.040). Conclusions: Pediatric CBF-AML was a unique setting of prognostic subtypes. Chemotherapy produced good responses. Additional chromosomal abnormalities was the only independent risk factor for OS in pediatric CBF-AML.

Comparision of visual quality after topography-guided and wavefront-optimized FS-LASIK in myopic eyes / 中华实验眼科杂志

Objective@#To compare the visual quality after topography-guided customized femtosecond laser-assisted in situ keratomileusis (FS-LASIK) and wavefront-optimized FS-LASIK treatment in myopic eyes.@*Methods@#A non-randomized controlled clinical study was performed.Seventy-eight eyes of 39 myopic patients undergoing FS-LASIK in Beijing Tongren Hospital from October 2016 to February 2017 were enrolled in this study and divided into two groups according to each patient's opinion, with matched demography between the two groups.Topography-guided customized FS-LASIK was performed on 42 eyes of 21 myopia in the topography-guided group, and wavefront-optimized FS-LASIK was performed on 36 eyes of 18 patients in the wavefront-optimized group.Visual acuity, refractive error, higher order aberrations (HOAs) and contrast sensitivity(CS) were compared between the two groups before and 6 months after surgery.Written informed consent was obtained from each patient before the operation.This study protocol was approved by Ethic Committee of Beijing Tongren Hospital (No.TRECKY2014-026).@*Results@#The postoperative uncorrected visual acuity (UCVA) levels were eaqual to or better than the preoperative best corrected visual acuity (BCVA) in 95.2% patients in the topography-guided group, and in 94.4% patients in the wavefront-optimized group 6 months after surgery, respectively.There were no significant differences in the spherical equivalent, sphere refraction and cylinder refraction between the two groups (all at P>0.05). The amount of induced coma was significantly lower in the topography-guided group than that in the wavefront-optimized group ([0.07±0.22]μm vs.[0.22±0.16]μm) at 6 mm pupil.LogCS improved under the 12.0 c/d in the background of mesopic in the topography-guided group and decreased under the 18.0 c/d both in the background of mesopic and mesopic + glare in the wavefront-optimized group 6 months after surgery, with signifcant differences between them (all at P<0.05). LogCS values under 6.0, 12.0 and 18.0 c/d in the background of mesopic and 18.0 c/d in the background of mesopic+ glare in the topography-guided group were significantly higher than those in the wavefront-optimized group 6 months after surgery, with significant differences between the two groups (all at P<0.05).@*Conclusions@#Topography-guided FS-LASIK has lower higher-order aberrations and better CS than wavefront-optimized FS-LASIK.

Study of clinical outcome and prognosis in pediatric core binding factor-acute myeloid leukemia / 中华血液学杂志

Objective@#To analyze the clinical outcome and the prognostic factor in pediatric patients with core binding factor-acute myeloid leukemia (CBF-AML).@*Methods@#A total of 121 newly diagnosed pediatric CBF-AML patients enrolled from Aug. 2005 to Sep. 2017 were retrospectively reviewed. Cumulative incidence of relapse (CIR), event-free survival (EFS) and overall survival (OS) rates were estimated by Kaplan-Meier method and prognostic factors were evaluated by Cox regression with SPSS.@*Results@#Of the 121 patients, 120 patients were assessed for bone marrow remission after induction chemotherapy. 100 cases (83.3%) achieved complete remission (CR) after the first course of chemotherapy. 119 cases (99.2%) achieved CR after the second course of chemotherapy. Of the 121 patients, 13 patients (10.7%) had recurrence with the median interval of recurrence as 13.8 months (3.7 to 58.8 months). 17 patients (14.0%) died. The CIR, EFS and OS at 3 years were 12.7%, 77.5% and 82.8%, respectively. The factors including age at diagnosis, sex, initial WBC count, presence of extramedullary leukemia, C-KIT expression, additional chromosomal abnormalities, and CR after the first course of chemotherapy were analyzed by multivariate regression analysis of Cox. Multivariate analysis identified that additional chromosomal abnormalities was the only independent risk factor affecting OS (HR=4.289, 95%CI 1.070-17.183, P=0.040).@*Conclusions@#Pediatric CBF-AML was a unique setting of prognostic subtypes. Chemotherapy produced good responses. Additional chromosomal abnormalities was the only independent risk factor for OS in pediatric CBF-AML.

Effect of long non-coding RNA Linc01296 on cisplatin resistance of ovarian cancer OVCAR-3 cells and its correlation to clinicopathological characteristics / 中国肿瘤生物治疗杂志

@# Objective: To investigate the effect of long non-coding RNA(lncRNA) Linc01296 on cisplatin (DDP) resistance in ovarian cancer cells and the mechanism. Methods: Fifty-three cases of ovarian cancer tissues, 22 cases of borderline ovarian tumor tissues and 17 cases of benign ovarian mass tissues were collected from Department of Gynecology, Sichuan Provincial People's Hospital during October 2017 to October 2018. The differential expression of Linc01296 mRNAin above tissue samples was detected by qPCR, and its relationship with clinicopathological data was analyzed. Human ovarian cancer cell line OVCAR-3 (OVCAR-3 Control) and its DDPresistant cell line (OVCAR-3 DR) were cultured. Lentiviral vectors expressing siRNA that targeting Linc01296 or siRNA control were transfected into OVCAR-3 DR cells respectively, as siLinc01296 group and siControl group. CCK-8 assay was used to detect DDP semi-inhibitory concentration (IC50) and drug resistance index of each group; real-time fluorescence quantitative PCR (qPCR) was used to detect the mRNA expressions of Linc 01296 and tumor stem cell related genes (Oct-4 and Sox-2) in cell lines of OVCAR-3 control, DR, DR siControl and siLinc 01296; and the protein expressions of Oct-4 and Sox-2 in each group were detected by Wb. Results: DDP IC50 and drug resistance index in OVCAR-3 DR group was significantly higher than that in Control group (all P <0.01), and DDP IC50 and drug resistance index in siLinc01296 group was significantly lower than that in DR siControl group (all P <0.01), but significantly higher than that in Control group (all P <0.01). The mRNA and protein expressions of Linc01296, Oct-4 and Sox-2 in DR group were significantly higher than that in Control group (all P <0.01), while those expressions in siLinc01296 group were significantly lower than those in DR siControl group ( P <0.01), and the mRNA and protein expressions of Oct-4 and Sox-2 in siLinc01296 group were higher than that in control group ( P <0.01). The expression of Linc01296 in ovarian cancer tissues was significantly higher than that in borderline ovarian tumor tissues and benign ovarian mass tissues ( P <0.01), which was positively correlated with lymph node metastasis and clinical stage ( P <0.05). Conclusion: Linc01296 can promote the occurrence of DDP resistance via promoting the expression of cancer stem cell markers in ovarian cancer; Linc01296 is highly expressed in ovarian cancer tissues and closely related to disease progression of patients.

Study of clinical outcome and prognosis in pediatric core binding factor-acute myeloid leukemia / 中华血液学杂志

Objective: To analyze the clinical outcome and the prognostic factor in pediatric patients with core binding factor-acute myeloid leukemia (CBF-AML). Methods: A total of 121 newly diagnosed pediatric CBF-AML patients enrolled from Aug. 2005 to Sep. 2017 were retrospectively reviewed. Cumulative incidence of relapse (CIR), event-free survival (EFS) and overall survival (OS) rates were estimated by Kaplan-Meier method and prognostic factors were evaluated by Cox regression with SPSS. Results: Of the 121 patients, 120 patients were assessed for bone marrow remission after induction chemotherapy. 100 cases (83.3%) achieved complete remission (CR) after the first course of chemotherapy. 119 cases (99.2%) achieved CR after the second course of chemotherapy. Of the 121 patients, 13 patients (10.7%) had recurrence with the median interval of recurrence as 13.8 months (3.7 to 58.8 months). 17 patients (14.0%) died. The CIR, EFS and OS at 3 years were 12.7%, 77.5% and 82.8%, respectively. The factors including age at diagnosis, sex, initial WBC count, presence of extramedullary leukemia, C-KIT expression, additional chromosomal abnormalities, and CR after the first course of chemotherapy were analyzed by multivariate regression analysis of Cox. Multivariate analysis identified that additional chromosomal abnormalities was the only independent risk factor affecting OS (HR=4.289, 95%CI 1.070-17.183, P=0.040). Conclusions: Pediatric CBF-AML was a unique setting of prognostic subtypes. Chemotherapy produced good responses. Additional chromosomal abnormalities was the only independent risk factor for OS in pediatric CBF-AML.