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Understanding the chemical composition and molecular transformation in soil dissolved organic matter (DOM) is important to the global carbon cycle. To address this issue, ultrahigh-resolution Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR-MS) was applied to investigate DOM molecules in 36 paddy soils collected from subtropical China. All the detected 7576 unique molecules were divided into seven compound groups, and nine trade-off relationships between different compound groups were revealed based on principal component analysis and Pearson's correlation. An optimized method was developed to evaluate all potential molecular transformations in DOM samples. The concept of thermodynamics was introduced to evaluate the identified molecular transformations and classify them as thermodynamically favorable (TFP) and thermodynamically limited (TLP) processes. Here, we first tried to understand the molecular trade-offs by using the potential molecular transformations. All the nine trade-offs could be explained by molecular transformations. Six trade-offs had bases of biochemical reactions, and the trade-off-related direct transformations could explain the content variations of carbohydrate-like, condensed aromatic-like, tannin-like, and lignin-like compounds in TLP. More reasonable explanations existed in the TLP rather than TFP, which demonstrated the critical role of external energy in the molecular transformation of soil DOM.
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Materia Orgánica Disuelta , Suelo , Ciclo del Carbono , China , Espectrometría de Masas , Suelo/químicaRESUMEN
CARM1 is a protein arginine methyltransferase (PRMT) that acts as a coactivator in a number of transcriptional programs. CARM1 orchestrates this coactivator activity in part by depositing the H3R17me2a histone mark in the vicinity of gene promoters that it regulates. However, the gross levels of H3R17me2a in CARM1 KO mice did not significantly decrease, indicating that other PRMT(s) may compensate for this loss. We thus performed a screen of type I PRMTs, which revealed that PRMT6 can also deposit the H3R17me2a mark in vitro CARM1 knockout mice are perinatally lethal and display a reduced fetal size, whereas PRMT6 null mice are viable, which permits the generation of double knockouts. Embryos that are null for both CARM1 and PRMT6 are noticeably smaller than CARM1 null embryos, providing in vivo evidence of redundancy. Mouse embryonic fibroblasts (MEFs) from the double knockout embryos display an absence of the H3R17me2a mark during mitosis and increased signs of DNA damage. Moreover, using the combination of CARM1 and PRMT6 inhibitors suppresses the cell proliferation of WT MEFs, suggesting a synergistic effect between CARM1 and PRMT6 inhibitions. These studies provide direct evidence that PRMT6 also deposits the H3R17me2a mark and acts redundantly with CARM1.
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Embrión de Mamíferos/enzimología , Fibroblastos/enzimología , Histonas/metabolismo , Mitosis , Proteína-Arginina N-Metiltransferasas/metabolismo , Animales , Histonas/genética , Metilación , Ratones , Ratones Noqueados , Proteína-Arginina N-Metiltransferasas/genéticaRESUMEN
PRMT5 is an arginine methyltransferase that accounts for the vast majority of the symmetric methylation in cells. PRMT5 exerts its function when complexed with MEP50/WDR77. This activity is often elevated in cancer cells and correlates with poor prognosis, making PRMT5 a therapeutic target. To investigate the PRMT5 signaling pathway and to identify genes whose loss-of-function sensitizes cancer cells to PRMT5 inhibition, we performed a CRISPR/Cas9 genetic screen in the presence of a PRMT5 inhibitor. We identified known components of the PRMT5 writer/reader pathway including PRMT5 itself, MEP50/WDR77, PPP4C, SMNDC1 and SRSF3. Interestingly, loss of PRMT1, the major asymmetric arginine methyltransferase, also sensitizes cells to PRMT5 inhibition. We investigated the interplay between PRMT5 and PRMT1, and found that combinatorial inhibitor treatment of small cell lung cancer and pancreatic cancer cell models have a synergistic effect. Furthermore, MTAP-deleted cells, which harbor an attenuated PRMT5-MEP50 signaling pathway, are generally more sensitive to PRMT1 inhibition. Together, these findings demonstrate that there is a degree of redundancy between the PRMT5 and PRMT1 pathways, even though these two enzymes deposit different types of arginine methylation marks. Targeting this redundancy provides a vulnerability for tumors carrying a co-deletion of MTAP and the adjacent CDKN2A tumor suppressor gene.
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Eliminación de Gen , Neoplasias/enzimología , Proteína-Arginina N-Metiltransferasas/metabolismo , Proteínas Represoras/metabolismo , Células A549 , Animales , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Etilenodiaminas/farmacología , Humanos , Isoquinolinas/farmacología , Células MCF-7 , Ratones Noqueados , Neoplasias/genética , Neoplasias/patología , Proteína-Arginina N-Metiltransferasas/antagonistas & inhibidores , Proteína-Arginina N-Metiltransferasas/genética , Pirimidinas/farmacología , Pirroles/farmacología , Proteínas Represoras/antagonistas & inhibidores , Proteínas Represoras/genética , Transducción de Señal/efectos de los fármacos , Transducción de Señal/genéticaRESUMEN
The coal pulverizing system is an important auxiliary system in thermal power generation systems. The working condition of a coal pulverizing system may directly affect the safety and economy of power generation. Prognostics and health management is an effective approach to ensure the reliability of coal pulverizing systems. As the coal pulverizing system is a typical dynamic and nonlinear high-dimensional system, it is difficult to construct accurate mathematical models used for anomaly detection. In this paper, a novel data-driven integrated framework for anomaly detection of the coal pulverizing system is proposed. A neural network model based on gated recurrent unit (GRU) networks, a type of recurrent neural network (RNN), is constructed to describe the temporal characteristics of high-dimensional data and predict the system condition value. Then, aiming at the prediction error, a novel unsupervised clustering algorithm for anomaly detection is proposed. The proposed framework is validated by a real case study from an industrial coal pulverizing system. The results show that the proposed framework can detect the anomaly successfully.
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BACKGROUND: Education for asthmatic children in the outpatient department is insufficient. AIM: To evaluate the efficacy of a nurse-led education pathway, a standard education programme, on children with asthma. METHODS: One hundred and eighty participants enrolled and were randomly assigned to either the control group or the intervention group. The intervention group received predetermined step-by-step education sessions based on the self-designed education pathway, while the control group received usual care. Asthma control, health-related quality of life, and health-care utilization measures were taken at baseline and at follow-up visits between February 2016 and May 2018. RESULTS: Significantly higher scores for health-related quality of life and inhaler technique at the third-month visit and asthma control test at the sixth-month visit were seen in the intervention group. The numbers of unscheduled physician visits and school absences were lower in the intervention group than in the control group within 6 months. However, no significant differences were observed in emergency department visits and hospitalizations. CONCLUSION: The nurse-led education pathway could be considered effective for children with asthma visiting the outpatient department.
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Asma/enfermería , Continuidad de la Atención al Paciente , Relaciones Enfermero-Paciente , Pacientes Ambulatorios , Asma/fisiopatología , Niño , Servicio de Urgencia en Hospital , Femenino , Hospitalización , Humanos , Masculino , Calidad de VidaRESUMEN
The type II arginine methyltransferase PRMT5 is responsible for the symmetric dimethylation of histone to generate the H3R8me2s and H4R3me2s marks, which correlate with the repression of transcription. However, the protein level of a number of genes (MEP50, CCND1, MYC, HIF1a, MTIF and CDKN1B) are reported to be downregulated by the loss of PRMT5, while their mRNA levels remain unchanged, which is counterintuitive for PRMT5's proposed role as a transcription repressor. We noticed that the majority of the genes regulated by PRMT5, at the posttranscriptional level, express mRNA containing an internal ribosome entry site (IRES). Using an IRES-dependent reporter system, we established that PRMT5 facilitates the translation of a subset of IRES-containing genes. The heterogeneous nuclear ribonucleoprotein, hnRNP A1, is an IRES transacting factor (ITAF) that regulates the IRES-dependent translation of Cyclin D1 and c-Myc. We showed that hnRNP A1 is methylated by PRMT5 on two residues, R218 and R225, and that this methylation facilitates the interaction of hnRNP A1 with IRES RNA to promote IRES-dependent translation. This study defines a new role for PRMT5 regulation of cellular protein levels, which goes beyond the known functions of PRMT5 as a transcription and splicing regulator.
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Ciclina D1/genética , Ribonucleoproteína Heterogénea-Nuclear Grupo A-B/genética , Sitios Internos de Entrada al Ribosoma , Proteína-Arginina N-Metiltransferasas/genética , Proteínas Proto-Oncogénicas c-myc/genética , ARN Mensajero/genética , Animales , Sitios de Unión , Línea Celular Transformada , Ciclina D1/metabolismo , Fibroblastos/citología , Fibroblastos/metabolismo , Regulación de la Expresión Génica , Células HEK293 , Células HeLa , Ribonucleoproteína Nuclear Heterogénea A1 , Ribonucleoproteína Heterogénea-Nuclear Grupo A-B/metabolismo , Humanos , Células MCF-7 , Metilación , Ratones , Unión Proteica , Biosíntesis de Proteínas , Proteína-Arginina N-Metiltransferasas/metabolismo , Proteínas Proto-Oncogénicas c-myc/metabolismo , ARN Mensajero/metabolismoRESUMEN
PRMT5 is the primary enzyme responsible for the deposition of the symmetric dimethylarginine in mammalian cells. In an effort to understand how PRMT5 is regulated, we identified a threonine phosphorylation site within a C-terminal tail motif, which is targeted by the Akt/serum- and glucocorticoid-inducible kinases. While investigating the function of this posttranslational modification, we serendipitously discovered that its free C-terminal tail binds PDZ domains (when unphosphorylated) and 14-3-3 proteins (when phosphorylated). In essence, a phosphorylation event within the last few residues of the C-terminal tail generates a posttranslational modification-dependent PDZ/14-3-3 interaction "switch." The C-terminal motif of PRMT5 is required for plasma membrane association, and loss of this switching capacity is not compatible with life. This signaling phenomenon was recently reported for the HPV E6 oncoprotein but has not yet been observed for mammalian proteins. To investigate the prevalence of PDZ/14-3-3 switching in signal transduction, we built a protein domain microarray that harbors PDZ domains and 14-3-3 proteins. We have used this microarray to interrogate the C-terminal tails of a small group of candidate proteins and identified ERBB4, PGHS2, and IRK1 (as well as E6 and PRMT5) as conforming to this signaling mode, suggesting that PDZ/14-3-3 switching may be a broad biological paradigm.
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Proteínas 14-3-3/metabolismo , Dominios PDZ , Proteína-Arginina N-Metiltransferasas/metabolismo , Proteínas 14-3-3/química , Secuencia de Aminoácidos , Animales , Línea Celular Tumoral , Membrana Celular/metabolismo , Humanos , Ratones , Fosforilación , Unión Proteica , Procesamiento Proteico-Postraduccional , Proteína-Arginina N-Metiltransferasas/químicaRESUMEN
Positive transcription elongation factor-b (P-TEFb) is required for the release of RNA polymerase II (RNAPII) from its pause near the gene promoters and thus for efficient proceeding to the transcription elongation. It consists of two core subunits-CDK9 and one of T-typed or K-typed cyclin, of which, cyclin T1/CDK9 is the major and most studied combination. We have previously identified a novel splice variant of cyclin T1, cyclin T1b, which negatively regulates the transcription elongation of HIV-1 genes as well as several host genes. In this study, we revealed the serine-arginine-rich protein, ASF/SF2, as a regulatory factor of the alternative splicing of cyclin T1 gene. ASF/SF2 promotes the production of cyclin T1b versus cyclin T1a and regulates the expression of cyclin T1-depedent genes at the transcription level. We further found that a cis-element on exon 8 is responsible for the skipping of exon 7 mediated by ASF/SF2. Collectively, ASF/SF2 is identified as a splicing regulator of cyclin T1, which contributes to the control of the subsequent transcription events. J. Cell. Biochem. 118: 4020-4032, 2017. © 2017 Wiley Periodicals, Inc.
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Empalme Alternativo/fisiología , Ciclina T/biosíntesis , Factores de Empalme Serina-Arginina/metabolismo , Línea Celular , Ciclina T/genética , Humanos , Isoformas de Proteínas/biosíntesis , Isoformas de Proteínas/genética , Factores de Empalme Serina-Arginina/genéticaRESUMEN
BACKGROUND Our research purpose was to compare the curative efficacy of different skin grafting methods for treating third-degree burn wounds. MATERIAL AND METHODS A total of 105 patients with third-degree burns were involved in this study. The burn wounds of these patients were treated using three different methods: Meek skin grafting, Stamp skin grafting, and Microskin grafting. Patients treated with different methods were placed in different groups. The skin graft survival rate, skin graft fusion time, wound healing time, total time of surgery, and 1% total body surface area (TBSA) treatment costs in each group were evaluated during and after the grafting procedures. After the operations, patients were followed up for 3 to 18 months in order to evaluate the postoperative outcomes. RESULTS The skin graft survival rate was significantly higher in the Meek group compared to the rates in the Stamp and Microskin groups (both P<0.01). In addition, the skin graft fusion time, wound healing time, and 1% TBSA treatment costs were significantly lower in the Meek group compared to those in the Stamp and Microskin groups (both P<0.01). Furthermore, the Meek group exhibited better results with respect to curative efficacy, scarring status, and joint activity in comparison to the other two groups (both P<0.05). CONCLUSIONS The Meek skin grafting method showed better clinical efficacy for treating large wound areas in third-degree burn patients compared to the Stamp and Microskin skin grafting methods.
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Quemaduras/cirugía , Trasplante de Piel/métodos , Adulto , Cicatriz/patología , Femenino , Estudios de Seguimiento , Supervivencia de Injerto , Humanos , Masculino , Persona de Mediana Edad , Periodo Posoperatorio , Piel/patología , Factores de Tiempo , Resultado del Tratamiento , Cicatrización de HeridasRESUMEN
UNLABELLED: Hepatitis B virus (HBV), a small enveloped DNA virus, chronically infects more than 350 million people worldwide and causes liver diseases from hepatitis to cirrhosis and liver cancer. Here, we report that hepatocyte nuclear factor 6 (HNF6), a liver-enriched transcription factor, can inhibit HBV gene expression and DNA replication. Overexpression of HNF6 inhibited, while knockdown of HNF6 expression enhanced, HBV gene expression and replication in hepatoma cells. Mechanistically, the SP2 promoter was inhibited by HNF6, which partly accounts for the inhibition on S mRNA. Detailed analysis showed that a cis element on the HBV genome (nucleotides [nt] 3009 to 3019) was responsible for the inhibition of the SP2 promoter by HNF6. Moreover, further analysis showed that HNF6 reduced viral pregenomic RNA (pgRNA) posttranscriptionally via accelerating the degradation of HBV pgRNA independent of La protein. Furthermore, by using truncated mutation experiments, we demonstrated that the N-terminal region of HNF6 was responsible for its inhibitory effects. Importantly, introduction of an HNF6 expression construct with the HBV genome into the mouse liver using hydrodynamic injection resulted in a significant reduction in viral gene expression and DNA replication. Overall, our data demonstrated that HNF6 is a novel host factor that can restrict HBV replication via both transcriptional and posttranscriptional mechanisms. IMPORTANCE: HBV is a major human pathogen whose replication is regulated by host factors. Liver-enriched transcription factors are critical for many liver functions, including metabolism, development, and cell proliferation, and some of them have been shown to regulate HBV gene expression or replication in different manners. In this study, we showed that HNF6 could inhibit the gene expression and DNA replication of HBV via both transcriptional and posttranscriptional mechanisms. As HNF6 is differentially expressed in men and women, the current results may suggest a role of HNF6 in the gender dimorphism of HBV infection.
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Replicación del ADN/genética , Regulación Viral de la Expresión Génica/genética , Virus de la Hepatitis B/metabolismo , Factor Nuclear 6 del Hepatocito/metabolismo , Caracteres Sexuales , Animales , Northern Blotting , Western Blotting , Línea Celular Tumoral , Cartilla de ADN/genética , Femenino , Vectores Genéticos/genética , Células HEK293 , Virus de la Hepatitis B/genética , Factor Nuclear 6 del Hepatocito/genética , Humanos , Inmunohistoquímica , Luciferasas , Masculino , Ratones , ARN Interferente Pequeño/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Transfección/métodosRESUMEN
A variety of cellular factors participates in the HIV-1 life cycle. Among them is the well characterized cyclin T1 (CYCT1). CycT1 binds to cyclin-dependent kinase 9 (CDK9) and forms the positive transcription elongation factor-b (P-TEFb). P-TEFb is then recruited by HIV-1 TAT to the HIV-1 long terminal repeat (LTR) promoter and subsequently leads to phosphorylation of the C-terminal domain of RNA polymerase II (pol II), enhanced processivity of pol II, and transcription of a full-length HIV-1 RNA. In this study, we report the identification of a new CYCT1 splice variant, designated as CYCT1b, and accordingly we renamed CYCT1 as CYCT1a. CYCT1b was detected in several cell lines, including primary human CD4 T cells, and its expression was subject to cell cycle regulation. Similar to CYCT1a, CYCT1b was primarily localized in the nucleus. CYCT1b expression was found to be inversely correlated with HIV-1 gene expression and replication. This inverse correlation appeared to involve TAT transactivation, CDK9 binding, and subsequent recruitment of P-TEFb to the HIV-1 LTR promoter and pol II C-terminal domain phosphorylation. In agreement with these findings, CYCT1b expression led to direct inhibition of TAT-transactivated transcription of the HIV-1 LTR promoter. Taken together, these results show that the newly identified CYCT1b splice variant inhibits HIV-1 transcription and may provide new clues for the development of anti-HIV strategies.
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Ciclina T/genética , Regulación Viral de la Expresión Génica , VIH-1/metabolismo , Transcripción Genética , Productos del Gen tat del Virus de la Inmunodeficiencia Humana/metabolismo , Empalme Alternativo , Secuencia de Aminoácidos , Secuencia de Bases , Ciclina T/metabolismo , Perfilación de la Expresión Génica , Células HEK293 , Células HeLa , Humanos , Células Jurkat , Datos de Secuencia Molecular , Sistemas de Lectura Abierta , Fosforilación , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , ARN Mensajero/metabolismo , Activación Transcripcional , Técnicas del Sistema de Dos HíbridosRESUMEN
Coronaviruses possess a cap structure at the 5' ends of viral genomic RNA and subgenomic RNAs, which is generated through consecutive methylations by virally encoded guanine-N7-methyltransferase (N7-MTase) and 2'-O-methyltransferase (2'-O-MTase). The coronaviral N7-MTase is unique for its physical linkage with an exoribonuclease (ExoN) harbored in nonstructural protein 14 (nsp14) of coronaviruses. In this study, the structure-function relationships of the N7-MTase were analyzed by deletion and site-directed mutagenesis of severe acute respiratory syndrome coronavirus (SARS-CoV) nsp14. The results showed that the ExoN domain is closely involved in the activity of the N7-MTase, suggesting that coronavirus N7-MTase is different from all other viral N7-MTases, which are separable from other structural domains located in the same polypeptide. Two of the 12 critical residues identified to be essential for the N7-MTase were located at the N terminus of the core ExoN domain, reinforcing a role of the ExoN domain in the N7-MTase activity of nsp14. The other 10 critical residues were distributed throughout the N7-MTase domain but localized mainly in the S-adenosyl-l-methionine (SAM)-binding pocket and key structural elements of the MTase fold of nsp14. The sequence motif DxGxPxA (amino acids [aa] 331 to 338) was identified as the key part of the SAM-binding site. These results provide insights into the structure and functional mechanisms of coronaviral nsp14 N7-MTase.
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Exorribonucleasas/química , Metiltransferasas/química , Caperuzas de ARN/metabolismo , ARN Viral/metabolismo , Síndrome Respiratorio Agudo Grave/virología , Coronavirus Relacionado al Síndrome Respiratorio Agudo Severo/enzimología , Proteínas no Estructurales Virales/química , Proteínas Virales/química , Exorribonucleasas/genética , Exorribonucleasas/metabolismo , Metilación , Metiltransferasas/genética , Metiltransferasas/metabolismo , Estructura Terciaria de Proteína , Caperuzas de ARN/genética , ARN Viral/genética , Coronavirus Relacionado al Síndrome Respiratorio Agudo Severo/química , Coronavirus Relacionado al Síndrome Respiratorio Agudo Severo/genética , Coronavirus Relacionado al Síndrome Respiratorio Agudo Severo/metabolismo , Proteínas no Estructurales Virales/genética , Proteínas no Estructurales Virales/metabolismo , Proteínas Virales/genética , Proteínas Virales/metabolismoRESUMEN
Soil organic carbon (SOC) mineralization is essential to biogeochemical recycling in terrestrial ecosystem. However, the microbial mechanisms underlying the nutrient-induced SOC mineralization remain uncertain. Here, we investigated how SOC mineralization was linked to microbial assembly processes as well as soil nutrient availability and stoichiometric ratio in a paddy rice ecosystem at four soil profile levels. Our results showed a sharp decrease in SOC mineralization from topsoil (112.61-146.34 mg CO2 kg-1 day-1) to subsoil (33.51-61.41 mg CO2 kg-1 day-1). High-throughput sequencing showed that both abundance and diversity of specialist microorganisms (Chao1: 1244.30-1341.35) significantly increased along the soil profile, while the generalist microorganisms (Chao1: 427.67-616.15; Shannon: 7.46-7.97) showed the opposite trend. Correspondingly, the proportion of deterministic processes that regulate specialist (9.64-21.59 %) and generalist microorganisms (21.17-53.53 %) increased and decreased from topsoil to subsoil, respectively. Linear regression modeling and partial least squares path modeling indicated that SOC mineralization was primarily controlled by the assembly processes of specialist microorganisms, which was significantly mediated by available soil C:N:P stoichiometry. This study highlighted the importance of soil stoichiometry-mediated bacterial community assembly processes in regulating SOC mineralization. Our results have an important implication for the integration of bacterial community assembly processes into the prediction of SOC dynamics.
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The coactivator associated arginine methyltransferase (CARM1) promotes transcription, as its name implies. It does so by modifying histones and chromatin bound proteins. We identified nuclear factor I B (NFIB) as a CARM1 substrate and show that this transcription factor utilizes CARM1 as a coactivator. Biochemical studies reveal that tripartite motif 29 (TRIM29) is an effector molecule for methylated NFIB. Importantly, NFIB harbors both oncogenic and metastatic activities, and is often overexpressed in small cell lung cancer (SCLC). Here, we explore the possibility that CARM1 methylation of NFIB is important for its transforming activity. Using a SCLC mouse model, we show that both CARM1 and the CARM1 methylation site on NFIB are critical for the rapid onset of SCLC. Furthermore, CARM1 and methylated NFIB are responsible for maintaining similar open chromatin states in tumors. Together, these findings suggest that CARM1 might be a therapeutic target for SCLC.
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Neoplasias Pulmonares , Carcinoma Pulmonar de Células Pequeñas , Animales , Ratones , Factores de Transcripción NFI , Proteína-Arginina N-Metiltransferasas/metabolismo , CromatinaRESUMEN
While opening-up promotes regional economic development, its impact on the residents' health level cannot be ignored. Based on provincial data of China from 2009 to 2020, the Gini Coefficient and Theil Index are used to analyze the regional inequalities in residents' health in China. The Difference-in-Difference model is constructed to study the impact of China's opening-up policies and other factors on residents' health. The results show that, firstly, the health levels of Chinese residents have steadily improved and regional inequalities have been gradually narrowing. Secondly, the Belt and Road Initiative has significantly improved the residents' health along the route, while the Pilot Free Trade Zone, which is another important opening-up policy in China, has had an inhibitory effect on the health of residents. Thirdly, it is proven that the Belt and Road Initiative improves the health of residents in provinces along the route by increasing the degree of opening-up and improving the regional environmental quality. This study will support and advance the UN's Sustainable Development Goals (SDGs), especially SDG3 (Good Health and Well-being) and SDG10 (Reduced Inequalities).
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Desarrollo Económico , Desarrollo Sostenible , ChinaRESUMEN
SPINDOC is tightly associated with the histone H3K4me3 effector protein SPIN1. To gain a better understanding of the biological roles of SPINDOC, we identified its interacting proteins. Unexpectedly, SPINDOC forms two mutually exclusive protein complexes, one with SPIN1 and the other with PARP1. Consistent with its ability to directly interact with PARP1, SPINDOC expression is induced by DNA damage, likely by KLF4, and recruited to DNA lesions with dynamics that follows PARP1. In SPINDOC knockout cells, the levels of PARylation are reduced, in both the absence and presence of DNA damage. The SPINDOC/PARP1 interaction promotes the clearance of PARP1 from damaged DNA, and also impacts the expression of known transcriptional targets of PARP1. To address the in vivo roles of SPINDOC in PARP1 regulation, we generate SPINDOC knockout mice, which are viable, but slightly smaller than their wildtype counterparts. The KO mice display reduced levels of PARylation and, like PARP1 KO mice, are hypersensitive to IR-induced DNA damage. The findings identify a SPIN1-independent role for SPINDOC in the regulation of PARP1-mediated PARylation and the DNA damage response.
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Proteínas Co-Represoras/metabolismo , Neoplasias/metabolismo , Poli(ADP-Ribosa) Polimerasa-1/metabolismo , Animales , Línea Celular , Daño del ADN , Reparación del ADN , Modelos Animales de Enfermedad , Humanos , Ratones , Ratones Noqueados , Neoplasias/genética , Neoplasias/patología , Dominios y Motivos de Interacción de ProteínasRESUMEN
Somatic mutations affecting CREBBP and EP300 are a hallmark of diffuse large B-cell lymphoma (DLBCL). These mutations are frequently monoallelic, within the histone acetyltransferase (HAT) domain and usually mutually exclusive, suggesting that they might affect a common pathway, and their residual WT expression is required for cell survival. Using in vitro and in vivo models, we found that inhibition of CARM1 activity (CARM1i) slows DLBCL growth, and that the levels of sensitivity are positively correlated with the CREBBP/EP300 mutation load. Conversely, treatment of DLBCLs that do not have CREBBP/EP300 mutations with CARM1i and a CBP/p300 inhibitor revealed a strong synergistic effect. Our mechanistic data show that CARM1i further reduces the HAT activity of CBP genome wide and downregulates CBP-target genes in DLBCL cells, resulting in a synthetic lethality that leverages the mutational status of CREBBP/EP300 as a biomarker for the use of small-molecule inhibitors of CARM1 in DLBCL and other cancers.
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Proteína de Unión a CREB/genética , Proteína p300 Asociada a E1A/genética , Histona Acetiltransferasas/metabolismo , Linfoma de Células B Grandes Difuso/genética , Linfoma de Células B Grandes Difuso/metabolismo , Proteína-Arginina N-Metiltransferasas/antagonistas & inhibidores , Mutaciones Letales Sintéticas/genética , Acetilación/efectos de los fármacos , Animales , Línea Celular , Regulación hacia Abajo/genética , Ratones , Ratones Endogámicos NOD , Ratones SCIDRESUMEN
The aberrant expression of protein arginine methyltransferase 5 (PRMT5) has been associated with multiple cancers. Using the proteolysis targeting chimera technology, we discovered a first-in-class PRMT5 degrader 15 (MS4322). Here, we report the design, synthesis, and characterization of compound 15 and two structurally similar controls 17 (MS4370) and 21 (MS4369), with impaired binding to the von Hippel-Lindau E3 ligase and PRMT5, respectively. Compound 15, but not 17 and 21, effectively reduced the PRMT5 protein level in MCF-7 cells. Our mechanism studies indicate that compound 15 degraded PRMT5 in an E3 ligase- and proteasome-dependent manner. Compound 15 also effectively reduced the PRMT5 protein level and inhibited growth in multiple cancer cell lines. Moreover, compound 15 was highly selective for PRMT5 in a global proteomic study and exhibited good plasma exposure in mice. Collectively, compound 15 and its two controls 17 and 21 are valuable chemical tools for exploring the PRMT5 functions in health and disease.
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Dipéptidos/farmacología , Inhibidores Enzimáticos/farmacología , Proteína-Arginina N-Metiltransferasas/antagonistas & inhibidores , Animales , Línea Celular Tumoral , Dipéptidos/síntesis química , Dipéptidos/farmacocinética , Diseño de Fármacos , Inhibidores Enzimáticos/síntesis química , Inhibidores Enzimáticos/farmacocinética , Humanos , Ligandos , Masculino , Ratones , Estructura Molecular , Proteína-Arginina N-Metiltransferasas/metabolismo , Proteolisis/efectos de los fármacos , Relación Estructura-Actividad , Proteína Supresora de Tumores del Síndrome de Von Hippel-Lindau/metabolismoRESUMEN
BACKGROUND: Asthma self-management education combining with behavior therapy is considered to be more effective. Goal setting is a common behavior change technique used to help patients self-manage their symptoms. However, empirical evidence around its effectiveness on asthma management lacks clarity. AIMS: To systematically integrate and appraise the evidence for effectiveness of goal setting interventions on asthma outcomes. METHODS: Databases included CENTRAL, PubMed, EMBASE, CINAHL and Proquest Psychology Database were systematically searched for relevant intervention studies employing goal setting technique as a method in asthma education program for self-management. Characteristic of studies and outcomes in clinical, psychosocial and healthcare utilization outcome were extracted. RESULTS: From a total of 2641 citations, 45 full-text articles were assessed for eligibility and 9 studies met the inclusion criteria. Eight studies were randomized controlled trial and one was before-after study. None studies have a high methodological quality. Goal-setting based intervention appeared to improve symptom control, quality of life and self-efficacy in adult patients with asthma. CONCLUSION: This systematic review highlighted the potential of a goal setting technique in the asthma self-management education. However, due to the limitations of the quality and quantity of the included literature, more rigorous studies are needed. In the future, better effective study protocol combining with goal setting approach and other behavior technique is needed to further investigate.
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AIMS: Paediatric pressure ulcers are a serious problem to healthcare service. Thus, effective and early identification of the risk of developing pressure ulcer is essential. The Braden Q scale is a widely used tool in the risk assessment of paediatric pressure ulcer, but its predictive power is controversial. Hence, we performed a meta-analysis to evaluate the predictive power of the Braden Q scale for pressure ulcer in hospitalised children and offer recommendations for clinical decision. METHODS: Studies that evaluated the predictive power of the Braden Q scale were searched through databases in English and Chinese, including Medline, Cochrane Library, Embase, CINAHL, SinoMed, CNKI, Wangfang and VIP. The studies were screened by two independent reviewers. QUADAS-2 was used to assess the risk of bias of eligible studies. Demographic data and predictive value indices were extracted. The pooled sensitivity, specificity and receiver operating characteristics (ROC) were calculated by MetaDiSc 1.4 using random-effects models. RESULTS: Cochran Qâ¯=â¯26.13 (Pâ¯=â¯0.0036) indicated the existence of heterogeneity; the I2 for pooled DOR was 61.7%, suggesting significant heterogeneity among the included studies. The pooled sensitivity and specificity were 0.73 (95% CI: 0.67-0.78) and 0.61 (95% CI: 0. 59-0.63), respectively, yielding a combined DOR of 3.47 (95% CI: 2-6.01). The area under the ROC curve was 0.7078 ± 0.0421, and the overall diagnostic accuracy (Q*) was 0.6591 ± 0.0337. Sensitivity analysis showed the results were robust. CONCLUSION: The Braden Q scale has moderate predictive validity with medium sensitivity and low specificity for pressure ulcers in hospitalised children. Further development and modification of this tool for use in paediatric population are warranted.