RESUMO
Elaeagnus pungens Thunb. is a common traditional Chinese herbal medicine. It has high medicinal, edible, and ornamental value. In Sep. 2020, a leaf spot disease was found on E. pungens in the campus of Nanjing Forestry University, China (31°36'51"N, 119°11'8"E). The incidence rate was ca. 77%. The disease primarily appeared as small brown spots on the leaves. Then, the spots enlarged and coalesced into regular or irregular gray necrotic lesions with dark margins. At the late stage of symptom development, black spots (acervulus) appeared on the necrotic lesions (Fig. S2A-C). Eight symptomatic leaves were collected and surface-sterilized using 75% ethanol for 30 s followed by 1% NaClO for 1 min, and then washed three times in sterile distilled water. Cuttings (ca. 5×5 mm) were made from the margins of the lesions and placed on 2% of potato dextrose agar (PDA) in Petri plates and incubated at 25 ± 2 °C for 5 days. The isolation frequency of pathogens from diseased tissues was ca. 100%. A total of four fungal isolates 3-3-1, 3-3-2, 3-3-3, and 3-3-4 were obtained using the monosporic isolation method and stored in the Forest Pathology Laboratory at Nanjing Forestry University. For molecular identification, the internal transcribed spacer (ITS), partial translation elongation factor 1-alpha (TEF1-α), and partial ß-tubulin (TUB2) were amplified from the isolate 3-3-1, 3-3-2, 3-3-3 and 3-3-4, with the corresponding primer sets published in Maharachchikumbura et al. (2014). The amplicons of ITS (ON510047, ON510048, ON510070, and ON510069), TEF1-α (ON808445, ON808446, ON808447, and ON808448), and TUB2 (ON808449, ON808450, ON808451, and ON808452) generated from the isolate 3-3-1, 3-3-2, 3-3-3, and 3-3-4 were sequenced and deposited in GenBank. The ITS, TEF1-α, and TUB2 of the isolate 3-3-1 shared the same nucleotide sequences with the corresponding sequences of the isolate 3-3-2, 3-3-3, and 3-3-4. The ITS, TEF1-α, and TUB2 sequences showed 100%, 97%, and 99% similarity to Neopestalotiopsis clavispora MFLUCC12-0281 (ex-type), respectively. Phylogenetic analysis using concatenated sequences of ITS, TEF1-α, and TUB2 also showed that isolate 3-3-1, 3-3-2, 3-3-3, and 3-3-4 clustered monophyletically with N. clavispora, and supported with a high bootstrap value (80%) (Fig. S1). Since these four isolates were same species based on phylogenetic analysis, isolate 3-3-4 was randomly chosen for the pathogenicity test and morphological analysis. Colonies of the isolate 3-3-4 grown on PDA were white, cottony, and flocculent, contained undulate edges with dense aerial mycelium on the surface, and averaged 12.2 mm d-1 growth at 25 °C (Fig. S2F). Black conidiomata formed superficially, scattered over the PDA at two weeks post incubation, 170.15-1820.32 × 90.33-1230.12 µm (n = 109), and contained slimy black conidial mass (Fig. S2G). Conidiogenous cells were pear-shaped to cylindrical, transparent, and colorless to pale yellow with smooth cell walls (Fig. S2H). Conidia were spindle shaped, five cells, four septa, 18.46-25.9 × 5.3-9.37 µm, (av ± SD = 23.31 ± 1.81 × 7.33 ± 1.07 µm, n = 34) (Fig. S2I). Apical and basal cells were lighter in color, mostly hyaline, and the middle three cells were darker in color, mostly brown. The apical cell showed two to three colorless, transparent unbranched accessory filaments, 9.68-30.59 µm in length, (av ± SD = 20.57 ± 4.52 µm, n = 95), whereas the basal cell only a single appendage, 3.52-9.4 µm in length, (av ± SD = 5.32 ± 1.29 µm, n = 34) (Fig. S2I). These morphological characteristics were similar to N. clavispora described by Daengsuwan et al. (2021). Based on phylogenetic analysis and morphological characteristics, isolate 3-3-1, 3-3-2, 3-3-3, and 3-3-4 were identified as N. clavispora. Healthy potted seedlings of E. pungens (63-85 cm in height, 0.7-1.6 cm in diameter) were selected for the pathogenicity test in vivo. The surface-sterilized leaves were wounded with sterilized needles (1 mm in dia.) and inoculated with mycelial plugs and conidial suspensions, respectively. One part of the leaves were inoculated with mycelial plugs (5 mm in dia.) of isolate 3-3-4. The other part of the leaves were inoculated with 10 µL of conidial suspensions (1×106 spores mL-1). The inoculated plants were kept in a growth chamber at ca. 25 ± 2 °C and ca. 90% RH under a 12-h photoperiod. PDA discs without fungi and sterilized dH2O were used as controls, respectively. All experiments were repeated twice, and each treatment had six replicates at least. After 10 and 12 days post-inoculation, the necrotic lesions appeared on the leaves inoculated with the mycelial plugs and conidial suspensions of the isolate 3-3-4, respectively (Fig. S2D and E). However, no lesions were found on the plants inoculated with PDA discs and dH2O (Fig. S2D and E). Fungal isolates were re-isolated from the infected leaves and shared similar morphological characteristics of colonies and conidia with the original one. Thus, Koch's postulates were fulfilled. Neopestalotiopsis clavispora was determined as the pathogens of a variety of plant diseases such as leaf spot on Taxus chinensis, gray blight on Camellia sinensis, and root and crown rot on strawberry (Kirschbaum et al., 2018; Wang et al., 2019a, b). To our knowledge, this is the first report of leaf spot caused by N. clavispora on E. pungens worldwide. The discovery will be helpful for monitoring and control of this disease in the future.
RESUMO
Alternative splicing is associated with human disease. In acute promyelocytic leukaemia (APL) patients with the long (L)-type promyelocytic leukaemia-retinoic acid receptor α fusion gene (PML-RARA), three alternative splicing isoforms can be detected: E5(+)E6(+), E5(-)E6(+), and E5(-)E6(-). This study is the first to demonstrate that alternative splicing of L-type PML-RARA is associated with time to achieve complete remission (CR) in APL. Higher expression of the E5(-)E6(-) isoform, the short isoform, was related to longer time to achieve CR. Each isoform was constructed into recombinant lentiviral vector and transfected into U937 cells. Compared with the E5(-)E6(+) and E5(+)E6(+) groups, the U937 cells with E5(-)E6(-) showed lower sensitivity to all-trans-retinoic acid treatment.
Assuntos
Antineoplásicos/farmacologia , Resistencia a Medicamentos Antineoplásicos/genética , Leucemia Promielocítica Aguda/genética , Proteínas de Fusão Oncogênica/genética , Isoformas de RNA , Tretinoína/farmacologia , Processamento Alternativo , Antineoplásicos/uso terapêutico , Linhagem Celular Tumoral , Ordem dos Genes , Humanos , Espaço Intracelular/metabolismo , Leucemia Promielocítica Aguda/tratamento farmacológico , Leucemia Promielocítica Aguda/metabolismo , Proteínas de Fusão Oncogênica/metabolismo , Transporte Proteico , Tretinoína/uso terapêutico , Células U937RESUMO
OBJECTIVES: We aimed to describe the characteristics and to compare the sex differences in the clinical features and prognosis of Chinese elderly patients with senile degenerative valvular heart disease (VHD). DESIGN: This study was a nationwide, multicentre, prospective cohort study. SETTING: Participants were enrolled consecutively from 69 hospitals nationwide in China from September to December 2016. PARTICIPANTS: A total of 2728 patients aged ≥60 years old with an aetiological diagnosis of moderate to severe degenerative VHD as defined by echocardiography were recruited. MAIN OUTCOME MEASURES: The baseline data and 1-year follow-up data were collected, and disease distribution, clinical features, treatment and prognosis were compared between different sex groups. RESULTS: Aortic disease was more common in men, and mitral disease was more common in women. Male patients were more likely to have smoking, coronary heart disease, cardiomyopathy, chronic obstructive pulmonary disease and coronary artery bypass grafting histories, while female patients had more hypertension and atrial fibrillation. The average age and left ventricular ejection fraction were significantly lower in men than in women (p<0.001), while the intervention rate (p=0.026) and total hospitalisation cost (p=0.016) of male patients were higher than those of female patients. There were no significant differences in perioperative complications, in-hospital outcomes or short and intermediate prognoses between the two groups. CONCLUSIONS: Currently, the intervention rate of elderly patients with VHD is still not ideal, with dominant factor-patient rejection. Heart failure was the critical reason for rehospitalisation. There were some differences between men and women in the distribution, severity, clinical characteristics and interventions in senile degenerative valvular disease.
Assuntos
Doenças das Valvas Cardíacas , Caracteres Sexuais , Idoso , Feminino , Doenças das Valvas Cardíacas/diagnóstico por imagem , Doenças das Valvas Cardíacas/epidemiologia , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Volume Sistólico , Função Ventricular EsquerdaRESUMO
Background: The epidemiology and management of valvular heart disease (VHD) have changed with economic development and population aging in China in recent decades. Objectives: This study sought to understand the distribution, etiology, and presentation and assess the current practice and outcomes of older patients with VHD in China. Methods: The authors conducted the first nationwide survey of older patients with VHD between September and December 2016 from 69 hospitals in 28 provinces and municipalities throughout China. Hospitalized patients over 60 years of age with moderate-to-severe VHD, infective endocarditis, or previous valvular intervention were consecutively enrolled. Results: Of 8,929 patients (median age of 69 years, 47.5% female), 8227 (92.1%) had native VHD. Mitral regurgitation was the most prevalent single VHD (26.9% of native VHD), followed by tricuspid regurgitation (16.5%), aortic regurgitation (10.6%), aortic stenosis (5.1%), and mitral stenosis (3.1%). Degenerative (37.2%), functional (21.8%), and rheumatic (15.0%) etiologies were the 3 most common causes. Among symptomatic patients with severe VHD, 37.3% underwent valvular intervention. The intervention rates decreased significantly with age across all types of VHD (P trend < 0.01). Valvular surgery covered 93.7% of interventions. The overall 1-year survival rate was 74.4% (95% CI: 63.4%-85.4%). Conclusions: This study provides a unique national insight into the contemporary spectrum and management of older VHD patients in China. With the increase in the health care demand, more resources and efforts are required for early detection, effective intervention, and targeting innovation on advanced therapeutic techniques and devices to improve the outcomes.
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JAK2V617F mutation has been reported in 90% of patients with polycythemia vera (PV) and about 50% of patients with essential thromobocythemia (ET) and primary myelofibrosis (PMF). Recently, acquired mutations in the transmembrane-juxtamembrane region of MPL (MPLW515 mutations) have been reported in approximately 5% of JAK2V617F-negative PMF and about 1% of all cases of ET. MPL is the receptor for thrombopoietin that regulates the production of platelets by bone marrow. It is likely that some mutations more closely related to ET in MPL exon10 may have been missed by current assays. We inferred that there might be other mutations in MPL exon10 for MPN patients in addition to MPLW515 mutations. To investigate its mutation types and prevalence in Chinese patients with myeloproliferative neoplasms (MPN), we performed mutation detection on MPL exon10 in 103 JAK2V617F-negative MPN patients by single strand conformation polymorphism (SSCP) and allele-specific PCR (AS-PCR) combined with sequencing. As a result, one previously unrecognized MPL mutation (12-bp in-frame insertion) was identified in one patient with ET in addition to an MPLW515K mutation identified in one PMF patient. This confirms our hypothesis that BCR/ABL negative and JAK2V617F-negative MPN patients have other mutations besides W515 mutation in MPL exon10 and mutations other than single nucleotide exchange also exist. In addition, MPL mutation was associated with Chinese MPN patients.
Assuntos
Éxons/genética , Janus Quinase 2/genética , Mutação/genética , Transtornos Mieloproliferativos/genética , Receptores de Trombopoetina/genética , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Sequência de Bases , China , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Adulto JovemRESUMO
BACKGROUND: N-terminal pro-B-type natriuretic peptide (NT-proBNP) may reflect early prognosis in patients with valvular heart disease (VHD). OBJECTIVES: The aim of this study was to examine the association between NT-proBNP and mortality in elderly patients with VHD. METHODS: A total of 5,983 elderly patients (age ≥60 years) with moderate or severe VHD underwent echocardiography and NT-proBNP measurement. VHD examined included aortic stenosis, aortic regurgitation, mitral stenosis, mitral regurgitation, tricuspid regurgitation, and multivalvular heart disease. NT-proBNP ratio was defined as measured NT-proBNP relative to the maximal normal values specific to age and sex. Disease-specific thresholds were defined on the basis of penalized splines and maximally selected rank statistics. RESULTS: The cohort had a mean age of 71.1 ± 7.6 years. At 1-year follow-up, 561 deaths (9.4%) had occurred. In penalized splines, relative hazards showed a monotonic increase with greater NT-proBNP ratio for death with different VHDs (p < 0.001 for all) except mitral stenosis. Higher NT-proBNP ratio, categorized by disease-specific thresholds, was independently associated with mortality (overall adjusted hazard ratio: 1.99; 95% confidence interval: 1.76 to 2.24; p < 0.001). Different subtypes of VHD all incurred excess mortality with elevated NT-proBNP ratio, with the strongest association detected for aortic stenosis (adjusted hazard ratio: 10.5; 95% confidence interval: 3.9 to 28.27; p < 0.001). The addition of NT-proBNP ratio to the prediction algorithm including traditional risk factors improved outcome prediction (overall net reclassification index = 0.28; 95% CI: 0.24 to 0.34; p < 0.001; likelihood ratio test p < 0.001). Results remained consistent in patients under medical care, with normal left ventricular ejection fractions, and with primary VHD. CONCLUSIONS: NT-proBNP provides incremental prognostic information for mortality in various VHDs. It could aid in risk stratification as a pragmatic and versatile biomarker in elderly patients.
Assuntos
Fatores Etários , Doenças das Valvas Cardíacas , Peptídeo Natriurético Encefálico/sangue , Fragmentos de Peptídeos/sangue , Medição de Risco/métodos , Idoso , Biomarcadores/sangue , Ecocardiografia/métodos , Feminino , Doenças das Valvas Cardíacas/sangue , Doenças das Valvas Cardíacas/diagnóstico , Doenças das Valvas Cardíacas/mortalidade , Doenças das Valvas Cardíacas/fisiopatologia , Humanos , Masculino , Mortalidade , Valor Preditivo dos Testes , Prognóstico , Modelos de Riscos Proporcionais , Fatores de Risco , Fatores SexuaisRESUMO
OBJECTIVE: To summarize clinical characteristics and treatment strategy of Chinese elderly mitral regurgitation (MR) inpatients under the current guidelines, and to identify factors related to treatment options in them. METHODS: A single center retrospective study was conducted in which patients hospitalized in Fuwai hospital from May 1st of 2014 to April 30 of 2015 with moderate to severe MR assessed by transthoracic echocardiography were enrolled consecutively (n = 1741). Patients > 60 years old were grouped as elderly group (n = 680) and patients < 60 years were grouped as control group (n = 1061). The elderly group was categorized into two subgroups based on surgical status. RESULT: s The mean age of the elderly group was 66.98 ± 5.94 years. The most common reason of MR in elderly group was degenerative MR (41.18%). Atherosclerotic risk factors such as hypertension, diabetes or hyperlipidaemia were more commonly observed in elderly group than the control group (45.44% vs. 25.17%, P < 0.001; 19.56% vs. 8.48%, P < 0.001; 35.29% vs. 19.51%, P < 0.001). Elderly group had higher EuroscoreIIscore (5.54 ± 2.42 vs. 3.15 ± 1.66), greater left ventricular end diastolic diameter (LVEDD) (57.72 ± 12.3 vs. 57.33 ± 10.19 mm) and a lower surgery rate (54.71% vs. 63.91%); P < 0.05. Age, left ventricular ejection fraction (LVEF), regurgitation grade, EuroScore-II high risk stratification and having diabetes were identified as factors associated with therapy decisions in elderly MR patients. CONCLUSIONS: Valve surgery was denied in 45.29% of elderly MR inpatients. Older age, impaired LVEF, lower regurgitation grade, EuroScore-II high risk stratification, and having diabetes were factors most significantly associated with surgery denial among elderly Chinese inpatients with MR.
Assuntos
Variação Genética , Leucemia Promielocítica Aguda/genética , Proteínas Nucleares/genética , Proteínas de Fusão Oncogênica/genética , Receptores do Ácido Retinoico/genética , Fatores de Transcrição/genética , Proteínas Supressoras de Tumor/genética , Processamento Alternativo , Sequência de Bases , Células da Medula Óssea/metabolismo , China , DNA/química , DNA/metabolismo , Éxons , Humanos , Íntrons , Leucemia Promielocítica Aguda/metabolismo , Proteínas Nucleares/química , Proteínas de Fusão Oncogênica/química , Proteína da Leucemia Promielocítica , RNA Mensageiro/química , RNA Mensageiro/metabolismo , Receptores do Ácido Retinoico/química , Receptor alfa de Ácido Retinoico , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Deleção de Sequência , Fatores de Transcrição/química , Translocação Genética , Proteínas Supressoras de Tumor/químicaRESUMO
The AML1 gene is an essential transcription factor regulating the differentiation of hematopoietic stem cells into mature blood cells. Though at least 12 different alternatively spliced AML1 mRNAs are generated, three splice variants (AML1a, AML1b and AML1c) have been characterized. Here, using the reverse transcription-polymerase chain reaction with outward-facing primers, we identified a novel non-polyadenylated transcript from the AML1 gene, with exons 5 and 6 scrambled. The novel transcript resisted RNase R digestion, indicating it is a circular RNA structure that may originate from products of mRNA alternative splicing. The expression of the novel transcript in different cells or cell lines of human and a number of other species matched those of the canonical transcripts. The discovery provides additional evidence that circular RNA could stably exist in vivo in human, and may also help to understand the mechanism of the regulation of the AML1 gene transcription.