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1.
BMC Genomics ; 24(1): 182, 2023 Apr 05.
Artículo en Inglés | MEDLINE | ID: mdl-37020265

RESUMEN

Agaricus bisporus is the most widely cultivated edible mushroom in the world with a only around three hundred years known history of cultivation. Therefore, it represents an ideal organism not only to investigate the natural evolutionary history but also the understanding on the evolution going back to the early era of domestication. In this study, we generated the mitochondrial genome sequences of 352 A. bisporus strains and 9 strains from 4 closely related species around the world. The population mitogenomic study revealed all A. bisporus strains can be divided into seven clades, and all domesticated cultivars present only in two of those clades. The molecular dating analysis showed this species origin in Europe on 4.6 Ma and we proposed the main dispersal routes. The detailed mitogenome structure studies showed that the insertion of the plasmid-derived dpo gene caused a long fragment (MIR) inversion, and the distributions of the fragments of dpo gene were strictly in correspondence with these seven clades. Our studies also showed A. bisporus population contains 30 intron distribution patterns (IDPs), while all cultivars contain only two IDPs, which clearly exhibit intron loss compared to the others. Either the loss occurred before or after domestication, that could suggest that the change facilitates their adaptation to the cultivated environment.


Asunto(s)
Agaricus , Genoma Mitocondrial , Agaricus/genética , Europa (Continente)
2.
Clin Exp Pharmacol Physiol ; 50(12): 944-953, 2023 12.
Artículo en Inglés | MEDLINE | ID: mdl-37688444

RESUMEN

Myocardial fibrosis (MF) is involved in hypertension, myocardial infarction and heart failure. It has been reported that circular RNA (circRNA) is a key regulatory factor of MF progression. In this study, we revealed that circ_0002295 and CXCR2 were elevated, and miR-1287 was reduced in MF patients. Knockdown of circ_0002295 effectively suppressed the proliferation, migration and MF progression. Circ_0002295 was the molecular sponge of miR-12878, and miR-1287 inhibitor reversed the biological functions of circ_0002295 on the myocardial fibrosis. CXCR2 was a target gene of miR-1287, and CXCR2 silencing relieved the impacts of miR-1287 inhibitor on cardiac myofibroblasts. Circ_0002295 promoted MF progression by regulating the miR-1287/CXCR2 axis, providing a possible circRNA-targeted therapy for MF.


Asunto(s)
Insuficiencia Cardíaca , MicroARNs , Infarto del Miocardio , Humanos , Línea Celular Tumoral , Proliferación Celular/genética , Corazón , MicroARNs/genética , Receptores de Interleucina-8B/genética , ARN Circular/genética
3.
Plant Dis ; 2023 Nov 03.
Artículo en Inglés | MEDLINE | ID: mdl-37923979

RESUMEN

Ampelopsis grossedentata, commonly known as "Vine Tea" and well-recognized for its rich flavonoid content, is mainly distributed in the southern regions of the Yangtze River basin in China. These regions include Hunan, Hubei, Jiangxi, and Guizhou provinces. Vine Tea is mainly consumed as an herbal tea and has garnered attention for its reported health benefits, including antioxidant, anti-inflammatory, anti-tumor, anti-diabetic, and neuroprotective properties. It has been used to alleviate coughs and sore throats (Zhang et al., 2021; Wang et al., 2017; Gao et al., 2009). In the Zhangjiajie region of Hunan province alone, the Vine Tea planting area reached 7,670.5 hectares and produced commercial goods worth 1.417 billion RMB in 2022. In May 2021, leaf margins and veins fading to yellowing mottling, and crumpling of leaf blades in the shape of a boat symptoms were found in ~16% of Vine Tea plants in the Sanjiakuan Township, Yongding District, Zhangjiajie region (29°15'E, 110°30' N) (Figure 1a, b, c). (Figure 1a, b, c). Phytoplasma-like microbial cells (small oval shaped bacterial cells, around 1000 nm in size) were observed in sieve tube cells in the phloem of diseased leaves using transmission electron microscopy. No such cell was observed in the phloem of healthy leaves (Figure 2a, b). To investigate the potential association between phytoplasma and the observed symptoms of the diseased plants, total DNA was isolated from ten diseasedeaves and compared with ten healthy leaves from the same field using SteadyPure Plant Genomic DNA Extraction Kit. The isolated DNAs were analyzed first in a direct PCR using universal phytoplasma primer pair R16mF2/R16mR1 targeting the 16S rRNA gene (Gundersen and Lee 1996) and specific pair rpF1/rpR1 (Lee et al. 1998) targeting the DNA fragment encoding partial ribosomal proteins (rp) L22 (complete) and S3 and S19 (partial). The initial amplified products were used as templates and further amplified by nested PCR respectively with primer pair R16F2n/R16R2 for the 16S rRNA gene (Lee et al. 1998) and the rpF2/rpR2 primer pair for the rp gene (Martini et al. 2007). No amplification was obtained with DNA from healthy leaf samples using any of the four primer pairs. The amplified fragments from diseased leaves by nested PCR were cloned and sequenced (Qingke Biotech, China). The obtained sequences have been deposited in GenBank with accession numbers OR282806 for the 16S rRNA gene and GenBank OR353012 for the rp gene. BLASTn analysis revealed that the partial 16S rRNA gene sequence in our sample shared 99.4% nucleotide sequence identity with 'Candidatus Phytoplasma sp.' (MW364378) and 'Peony yellows phytoplasma' (KY814723) of the 16SrI group. Similarly, our rp gene sequence shared 99.6% nucleotide identity with the rpI group of phytoplasma such as the 'Balsamine virescence phytoplasma' (JN572890) and 'Paulownia witches'-broom phytoplasma' (HM146079). Phylogenetic analysis of the 16S rRNA and rp sequences using MEGA version 7.0 revealed that the phytoplasma strain associated with A. grossedentata yellow leaf syndrome in our study site belonged to the 16SrI (Candidatus Phytoplasma asteris) group of phytoplasma (Figure 3a, b). Using the interactive online phytoplasma classification tool iPhyClassifier (Zhao et al., 2009), virtual restriction fragment length polymorphism (RFLP) analysis of the 16S rRNA gene sequences showed our strain having a distinct RFLP map but was closest to that of the onion yellow phytoplasma 16SrI-B subgroup (GenBank accession number: AP006628), with a similarity coefficient of 0.94 (Figure 4a, b). To confirm phytoplasma transmission, healthy plants were inoculated with three scions of infected plants of A. grossedentata. After 16 days, the new leaves of the inoculated A. grossedentata showed yellow leaf symptoms (Figure 5a, b, c), akin to the symptoms originally observed in the field, and the outer contour of the leaf margin appeared chlorotic. After 26 days, primer pairs R16mF2/R16R1 and R16F2n/R16R2 were used for nested PCR detection of phytoplasma in symptomatic A. grossedentata leaves. Phytoplasma was detected in the first and second leaves of symptomatic branches and leaves while negative control showed no amplification. Sequencing of the amplified fragments showed 100% nucleotide identity to the strain from the grafting source. Our results indicated that the pathogen and the disease can be transmitted by tissue grafting, consistent with the biological characteristics of phytoplasma, and further confirmed that the phytoplasma was the pathogen of yellow leaf syndrome of A. grossedentata. Toour knowledge, this is the first report of phytoplasma of group 16SrI affecting A. grossedentata.

4.
BMC Cancer ; 22(1): 909, 2022 Aug 20.
Artículo en Inglés | MEDLINE | ID: mdl-35986311

RESUMEN

BACKGROUND: HuR/ELAVL1 (embryonic lethal abnormal vision 1) was a downstream target of miR-29b in some cancer cells. HuR protein exerts important prognostic effects of involving in the pathogenesis and development of acute myeloid leukemia (AML). This study aims to investigate the role of miR-29b-3p in biological behaviors of AML cells by targeting HuR and the involvement of the NF-κB and JAK/STAT signaling pathways. METHODS: The expressions of HuR and miR-29b-3p in AML cells were determined using RT-qPCR and Western blot, and the association between them was analyzed using the Spearman method. Next, the target relationship between HuR and miR-29b-3p was predicted by biological information databases and verified by the dual-luciferase reporter gene assay. MTS, methyl cellulose, flow cytometry and transwell assay were employed to detect the cell proliferation, clone formation, cell cycle and apoptosis, invasion and migration respectively, the effect of miR-29b-3p targeted HuR on the biological behaviors of AML cells was explored after over- /down-expression of miR-29b-3p and rescue experiment. Then, immunofluorescence assay and western blot were employed to detect location expression and phosphorylation levels of NF-κB and JAK/STAT signaling pathways related molecules respectively. RESULTS: HuR was negatively correlated with miR-29b-3p, and was the downstream target of miR-29b-3p in AML cells. When miR-29b-3p was overexpressed in AML cells, HuR was down-regulated, accompanied by cell viability decreased, cell cycle arrest, apoptosis increased, invasion and migration weakened. Moreover, the opposite result appeared after miR-29b-3p was down-regulated. The rescue experiment showed that miR-29b-3p inhibitor could reverse the biological effect of HuR down-regulation in AML cells. Molecular pathway results showed that miR-29b-3p could inhibit p65 expression in nucleus and phosphorylation levels of p65, IκBα, STAT1, STAT3 and STAT5. CONCLUSION: miR-29b-3p can inhibit malignant biological behaviors of AML cells via the inactivation of the NF-κB and JAK/STAT signaling pathways by targeting HuR. miR-29b-3p and its target HuR can be used as a new potential molecular for AML treatment.


Asunto(s)
Leucemia Mieloide Aguda , MicroARNs , Apoptosis/genética , Línea Celular Tumoral , Proliferación Celular/genética , Humanos , Leucemia Mieloide Aguda/patología , MicroARNs/genética , MicroARNs/metabolismo , FN-kappa B/genética , FN-kappa B/metabolismo , Transducción de Señal/genética
5.
BMC Microbiol ; 21(1): 75, 2021 03 06.
Artículo en Inglés | MEDLINE | ID: mdl-33676418

RESUMEN

OBJECTIVE: The aim of this study is to evaluate the efficacy of the strain Paenibacillus polymyxa HX-140, isolated from the rhizosphere soil of rape, to control Fusarium wilt of cucumber seedlings caused by Fusarium oxysporum f. sp. cucumerinum. RESULTS: Strain HX-140 was able to produce protease, cellulase, ß-1,3-glucanase and antifungal volatile organic compounds. An in vitro dual culture test showed that strain HX-140 exhibited broad spectrum antifungal activity against soil-borne plant pathogenic fungi. Strain HX-140 also reduced the infection of Fusarium wilt of cucumber seedlings by 55.6% in a greenhouse pot experiment. A field plot experiment confirmed the biocontrol effects and further revealed that antifungal activity was positively correlated with inoculum size by the root-irrigation method. Here, inoculums at 106 107 and 108 cfu/mL of HX-140 bacterial suspension reduced the incidence of Fusarium wilt of cucumber seedling by 19.5, 41.1, and 50.9%, respectively. CONCLUSIONS: Taken together, our results suggest that P. polymyxa HX-140 has significant potential in the control of Fusarium wilt and possibly other fungal diseases of cucumber.


Asunto(s)
Agentes de Control Biológico , Cucumis sativus/microbiología , Fusarium/fisiología , Interacciones Microbianas/fisiología , Paenibacillus polymyxa/fisiología , Enfermedades de las Plantas/prevención & control , Brassica napus/microbiología , Enfermedades de las Plantas/microbiología , Plantones/microbiología , Microbiología del Suelo
6.
Cardiology ; 146(6): 690-697, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34464951

RESUMEN

BACKGROUND: We recently developed the Coronary Artery Tree description and Lesion EvaluaTion (CatLet) angiographic scoring system. Our preliminary study demonstrated that the CatLet score better predicted clinical outcomes than the SYNTAX score. The current study aimed at assessing whether 3 clinical variables (CVs) - age, serum creatinine, and left ventricular ejection fraction (LVEF) - improved the performance of the CatLet score in outcome predictions in patients with acute myocardial infarction (AMI). METHODS: This study was a post hoc study of the CatLet score validation trial. Primary endpoint was major adverse cardiac or cerebrovascular events (MACCEs), and secondary endpoints were all-cause deaths and cardiac deaths. RESULTS: Over 1,185 person-years (median [interquartile range], 4.3 [3.8-4.9] years), there were 64 MACCEs (20.8%), 56 all-cause deaths (18.2%), and 47 cardiac deaths (15.2%). The addition of the 3 CVs to the stand-alone CatLet score significantly increased the Harrell's C-index by 0.0967 (p = 0.002) in MACCEs, by 0.1354 (p < 0.001) in all-cause deaths, and by 0.1187 (p = 0.001) in cardiac deaths. When compared with the stand-alone CatLet score, improved discrimination and better calibration led to a significantly refined risk stratification, particularly at the intermediate-risk category. CONCLUSIONS: CatLet score had a predicting value for clinical outcome in AMI patients. This predicting value can be improved through a combination with age, serum creatinine, and LVEF (http://www.chictr.org.cn; unique identifier: ChiCTR-POC-17013536).


Asunto(s)
Creatinina/sangre , Infarto del Miocardio , Factores de Edad , Estudios de Seguimiento , Humanos , Infarto del Miocardio/diagnóstico por imagen , Volumen Sistólico , Función Ventricular Izquierda
7.
J Clin Lab Anal ; 35(8): e23817, 2021 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-34114685

RESUMEN

BACKGROUND: Doxorubicin is a first-line chemotherapy agent on human myelogenous leukemia clinical treatment, but the development of chemoresistance has largely limited curative effect. In this study, we aimed to evaluate the biological function and molecular mechanisms of CrkL to Doxorubicin resistance. METHODS: Quantitative reverse transcription-PCR (qRT-PCR) assay was performed to examine the expression of CrkL in K562 and K562/ADR cells. The expression of CrkL was silenced through RNA interference technology. MTT assay and flow cytometry were performed to detect the proliferation inhibition and apoptosis rate after CrkL siRNA transfection. The protein expression changes of PI3K/AKT/MRP1 pathway induced by CrkL siRNA were observed by Western Blot assay. Xenograft tumor model was carried out to observe tumor growth in vivo. RESULTS: We observed that silencing of CrkL could effectively increase apoptosis rate induced by doxorubicin and dramatically reversed doxorubicin resistance in K562/ADR cells. Further studies revealed knockdown CrkL expression suppressed PI3K/Akt/MRP1 signaling, which indicated CrkL siRNA reversed doxorubicin effect through regulating PI3K/Akt/MRP1 pathway. In addition, overexpression of MRP1 could evidently reduce apoptosis rate and reversed the inhibitory effects of doxorubicin resistance caused by CrkL siRNA on K562/ADR cells. Finally, in vivo experiments revealed that CrkL silencing acted a tumor-suppressing role in myelogenous leukemia via regulating PI3K/Akt/MRP1 signaling. CONCLUSION: Together, we indicated that CrkL is up-regulated in myelogenous leukemia cells and silencing of CrkL could reverse Doxorubicin resistance effectively. These results show a potential novel strategy for intervention chemoresistance in myelogenous leukemia during chemotherapy.


Asunto(s)
Proteínas Adaptadoras Transductoras de Señales/genética , Doxorrubicina/farmacología , Resistencia a Antineoplásicos/genética , Animales , Antibióticos Antineoplásicos/farmacología , Apoptosis/efectos de los fármacos , Apoptosis/genética , Resistencia a Antineoplásicos/efectos de los fármacos , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Silenciador del Gen , Humanos , Células K562 , Ratones Desnudos , Proteínas Asociadas a Resistencia a Múltiples Medicamentos/genética , Proteínas Asociadas a Resistencia a Múltiples Medicamentos/metabolismo , Fosfatidilinositol 3-Quinasas/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Transducción de Señal/efectos de los fármacos , Transducción de Señal/genética , Ensayos Antitumor por Modelo de Xenoinjerto
8.
Plant Dis ; 2021 Feb 22.
Artículo en Inglés | MEDLINE | ID: mdl-33616430

RESUMEN

Lilium lancifolium Thunb., commonly known as Juandan lily and tiger lily, is widely cultivated in China for its edible bulbs and medicinal properties, with a commercial value worth of ~RMB 6 billion Yuan per year. Bulb rot is an increasingly common disease on L. lancifolium, significantly impacting both the quantity and quality of the main product, the scaled bulbs. Typically, the causal pathogens invade the plant through wounds in the root or the ends of the bulb, causing the roots and bulb to brown and rot, which can eventually lead to stem wilt and death of the whole plants. During pathogenesis, the infected bulbs typically turn from white to brown, with sunken lesions and later the scales flaking off from the base of the bulb (Figure 1A and 1B). Plants growing from infected bulbs are generally short, with discolored leaves, wilting, and death at an early stage. Bulb rot is commonly observed in fields with excess water and a history of continuous Juandan lily cultivation. For this study, wilted L. lancifolium plants with rotted bulbs were collected from Longshan in Hunan, Enshi in Hubei, Yixing in Jiangsu, and Lu'an in Anhui in 2018 and 2019. Infected bulbs were surface sterilized with 75% ethanol for 30 seconds, followed by disinfection with 2% sodium hypochlorite for 5 minutes, and then rinsing with sterile water three times. The surface-sterilized tissue was divided into small pieces of 0.5 × 0.5 cm in size, placed on potato dextrose agar (PDA) medium containing 50 mg/l streptomycin sulfate, and incubated at 25℃. Mycelia growing from diseased tissues were sub-cultured onto fresh PDA medium to obtain pure culture, which formed dense white hyphae after a few days (Figure 1C and 1D). Colonies on PDA produced abundant condia about 15 days after subculturing. Microconidia were abundant, solitary, thin walled, hyaline, ovoid, 0 to 1 septate, with an average size of 6.1 × 2.6 µm (n=50) (Figure 1E). Macroconidia had a curved apical cell and foot-like basal cell with 3 to 5 septa, with an average size of 35.4 × 4.3 µm (n=30) (Figure 1E). No chlamydospore was observed. These morphological characteristics of the causal pathogen were similar to those of Fusarium spp. (Leslie et al., 2006). To identify the Fusarium isolates to species level, DNA fragments of the internal transcribed spacer (ITS) regions of the ribosomal RNA gene cluster, translation elongation factor subunit 1-alpha (TEF1-α), and RNA polymerase II subunit 2 (RPB2) genes were amplified using primers ITS1/ITS4, EF1/EF2, and 7cF/11aR respectively and sequenced (Choi et al. 2018; Jiang et al. 2018; Choi et al. 2017). BLAST analyses showed that the ITS (GenBank Accession No. MT549849), TEF1-α (GenBank Accession No. MT553348), and RPB2 (Accession No. MW201686) sequences of our isolates shared the highest sequence identities (98-100%) with those of F. fujikuroi reference strains in GenBank. A phylogenetic tree showing the relationship between one of our strains, S106, and those of the closely related species within the F. fujikuroi species complex was constructed by the maximum likelihood method using MEGA X (Kumar et al. 2018) (Figure 2). Based on the morphological characteristics and DNA sequences, the strains were identified as F. fujikuroi sensu stricto. We used two methods, an ex vivo assay using Juandan lily bulb scales and an in vivo assay using potted Juandan lily plants, to confirm pathogenicity for one representative F. fujikuroi strain from each of the four geographic regions to fulfill Koch's postulates (Bian et al. 2016; Zeng et al. 2019). In the ex vivo assay, actively growing mycelia on PDA plates were cut into 5mm diameter fungal blocks as inocula. To prepare healthy Juandan lily bulb scales as test tissues, healthy fresh scales were first surface sterilized using 75% alcohol for 30 seconds, followed by treatment of 2% sodium hypochlorite for 5 minutes, and then rinsed with sterile water 3 times. The scales were punctured with sterilized dissecting needles, the 5mm mycelial blocks containing the PDA medium were then inoculated on the punctured wound of the scales. Sterile PDA culture medium without mycelia was inoculated on the punctured wound as a negative control. After inoculation, Juandan lily scales were placed in sterile culture dishes with two layers of sterilized filter paper and 5ml of sterile water in each dish. Six Juandan lily scales were placed in each dish, with different treatments placed in different dishes, and the dishes were placed in an incubator in the dark at 25℃. After 10 days of incubation, we found that the F. fujikuroi-inoculated Juandan lily bulb scales showed disease symptoms (brownish lesion) similar to those in the original field collected infected bulb samples (Figure 1F). However, such symptoms were not observed in the negative control group. The pathogenicity test was performed 3 times for each isolate, each with six repeats. In the in vivo pathogenicity test using potted lily plants, we prepared actively growing cultures of our F. fujikuroi strains by incubating them in a liquid medium, the potato dextrose broth, for 3 days in a shaker-incubator at 25℃ and 180rpm. The asexual spores conidia from the fungal cultures were harvested by filtration through eight layers of sterile cheese clothes and with spore concentrations adjusted to 1×107 conidia per ml. Healthy Juandan lily bulbs were selected and one bulb was planted in each pot containing sterilized soil. Each pot was inoculated with 1ml conidia suspension, at the base soil where the bulbs were planted. The pots were placed in a growth chamber at 25℃ with a 12 h light and 12 h dark cycle. Symptoms similar to those observed in diseased bulbs in the field were observed, with symptoms at 30 days after inoculations shown in Figure 3. Specifically, most of the roots, bulb plate and scale tissues of Juandan lily plants inoculated with F. fujikuroi conidia were rotten and turned black, with few new roots. In addition, the infected plants showed stunted growth (Figure 3). In contrast, the uninoculated plants grew normally, with dense new roots and healthy-looking bulbs, and no rot symptom (Figure 3). The fungi were re-isolated from the infected Juandan lily tissues from both pathogenicity assays, following the procedures described above for isolating and identifying the fungal cultures from infected field samples. These re-isolated fungi were shown to have colony morphology and DNA sequences at the three loci identical to those of our inoculated F. fujikuroi strains. Several Fusarium species have been reported as pathogens of lily plants in China, including F. oxysporum, F. solani and F. tricinctum (Li, et al., 1995; Li, et al., 2013). In addition, F. redolens has been reported previously in ornamental lily in Ukraine (Zerova, 1940). Indeed, Fusarium moniliforme, one of the disused synonyms of F. fujikuroi (Seifert et al. 2003), has been reported as a causal agent for diseases in lily. However, it's now known that the originally defined F. fujikuroi sensu lato is in fact a large species complex consisting of over 60 recognized species, including F. fujikuroi sensu stricto (Moussa et al. 2017; Choi et al. 2018). In addition, there are over 100 species in the genus Lilium as well as many other species with their common names including the word "lily" but are not in the Lilium genus. To our knowledge, this is the first confirmed report of bulb rot of Juandan lily L. lancifolium caused by F. fujikuroi sensu stricto in China. Our result should help with future monitoring and control of Juandan lily diseases.

9.
Int J Syst Evol Microbiol ; 70(2): 1178-1185, 2020 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-31860431

RESUMEN

During a survey of endophytic fungi in plant roots in secondary forests in Yunnan, China, a novel ascomyceteous taxon, Beltrania sinensis, was isolated from Quercus cocciferoides Hand.-Mazz. and Fraxinus malacophylla Hemsl. This novel species is characterized by having oval or obovoid conidiogenous cells with several apical, flat-tipped denticles, and biconic, aseptate, smooth, pale brown conidia with a hyaline to subhyaline equatorial transverse band and apical tubular appendage. Phylogenetic analysis of the combined sequences of the internal transcribed spacer and the LSU rRNA gene confirmed its novel species status within the genus Beltrania. Here, the novel species is described and illustrated, and a taxonomic key to species in the genus Beltrania is provided.


Asunto(s)
Filogenia , Raíces de Plantas/microbiología , Quercus/microbiología , Xylariales/clasificación , China , ADN de Hongos/genética , ADN Espaciador Ribosómico/genética , Técnicas de Tipificación Micológica , Análisis de Secuencia de ADN , Esporas Fúngicas , Xylariales/aislamiento & purificación
10.
Yi Chuan ; 42(6): 556-564, 2020 Jun 20.
Artículo en Zh | MEDLINE | ID: mdl-32694114

RESUMEN

The CRISPR/Cas system is the most popular genome editing technology in recent years and has been widely used in crop improvement. Compared with introducing the CRISPR/Cas system into plant cells with DNA constructs, introducing CRISPR/Cas ribonucleoprotein (RNP) to perform genome editing excels in rapid action, low off-target rates and is free of DNA insertions in editing plants. However, efficient delivery of CRISPR/Cas RNP into plant cells and achieving high editing frequency are still very challenging, which limits the extensive implementation of CRISPR/Cas RNP-mediated genome editing in plants. In this review, we summarize the progress of protein and RNP delivery methods in plant cells, and provide new perspectives of further development and future applications of the CRISPR/Cas RNP technology in plant genome editing.


Asunto(s)
Sistemas CRISPR-Cas , Repeticiones Palindrómicas Cortas Agrupadas y Regularmente Espaciadas , Edición Génica , Sistemas CRISPR-Cas/genética , Genoma de Planta/genética , Plantas , Ribonucleoproteínas/genética
11.
Yi Chuan ; 40(12): 1075-1091, 2018 Dec 20.
Artículo en Zh | MEDLINE | ID: mdl-30559097

RESUMEN

The life science has entered a new chapter with the revolutionary implementation of the CRISPR/Cas9 genome editing technology in various living organisms. With the unique flexibility, feasibility and extendibility, the CRISPR/Cas9 technology greatly accelerates genetic engineering research, as well as plant molecular breeding. However, it has become a challenge to screen for and identify genome-edited plants at early stages in a rapid and high-throughput fashion, due to the massive number of plants produced from transformation process. In this review, we summarize the molecular methods developed in recent years to identify genome-edited plants. We compare their advantages and disadvantages, and the scope of application. In addition, we provide insights of the development trend of detection methods for plant genome editing. This review will serve as a reference for future genome editing research in plants.


Asunto(s)
Sistemas CRISPR-Cas , Edición Génica , Ingeniería Genética , Plantas/genética , Genoma de Planta
12.
Acta Pharmacol Sin ; 38(10): 1394-1400, 2017 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-28552912

RESUMEN

Resveratrol has long been known as an antioxidant and a chemopreventive agent. Similar to resveratrol, pterostilbene (PT) is also a phenolic compound extracted from the Vitis species. However, there are few studies on the antitumor effect of PT. Thus, we investigated the effects of PT on the endometrial cancer (EC) cells in vitro and the related molecular mechanisms. Treatment of EC cell lines HTB-111 and Ishikawa with PT (25-100 µmol/L) dose-dependently suppressed the cell viability and induced apoptosis. Using miR microarrays, we examined the miR expression profile in Ishikawa cells with or without PT, and revealed that miR-663b was the most decreased in PT-treated Ishikawa cells. Furthermore, we predicted and verified that the pro-apoptosis factor BCL2L14 is the direct target of miR-663b. Over-expression of miR-663b and knock-down of BCL2L14 counteracted the suppressing effects of PT on HTB-111 and Ishikawa cells. In addition, we evaluated the miR-663b levels in EC tissues of 51 patients using an in situ hybridization technique. With the median of the score of miR-663b as a cut-off value, these EC patients were divided into two groups, and the patients with high miR-663b expression had significantly poor prognosis.


Asunto(s)
Antineoplásicos Fitogénicos/farmacología , Neoplasias Endometriales/tratamiento farmacológico , MicroARNs/genética , Estilbenos/farmacología , Antineoplásicos Fitogénicos/administración & dosificación , Antineoplásicos Fitogénicos/aislamiento & purificación , Apoptosis/efectos de los fármacos , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Regulación hacia Abajo/efectos de los fármacos , Neoplasias Endometriales/patología , Femenino , Técnicas de Silenciamiento del Gen , Humanos , Pronóstico , Proteínas Proto-Oncogénicas c-bcl-2/genética , Estilbenos/administración & dosificación , Estilbenos/aislamiento & purificación , Vitis/química
13.
Med Sci Monit ; 23: 3064-3070, 2017 Jun 23.
Artículo en Inglés | MEDLINE | ID: mdl-28642455

RESUMEN

BACKGROUND Mediastinal diseases are difficult to diagnose due to diverse origins and complex anatomical structure of the mediastinal tissues. The prospective study aimed to compare the diagnostic efficiency of video-assisted mediastinoscopy (VAM) and endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) for mediastinal lesions without pulmonary abnormalities. MATERIAL AND METHODS We divided 100 mediastinal lymphadenectasis patients without pulmonary abnormalities into a VAM group and an EBUS group. The pathological results of each group were regarded as the endpoints. SPSS19.0 statistical software was used. RESULTS The diagnostic accuracy, sensitivity, and specificity of VAM were 96%, 97.4%, and 100%, respectively; those of EBUS-TBNA diagnosis were 62%, 87.1%, and 100%, respectively. There was a statistically significant difference in the diagnostic sensitivity of benign mediastinal lesions between the 2 groups (P<0.01). Compared with the EBUS group (62%), the accuracy in the VAM group was significantly higher (96%) (P<0.01). CONCLUSIONS We found that the diagnostic accuracy of VAM for mediastinal lymphadenectasis without pulmonary abnormalities is superior to that of EBUS. Therefore, for patients with mediastinal lymphadenectasis or mediastinal mass and without pulmonary abnormalities, mediastinoscopy is recommended as the first choice.


Asunto(s)
Biopsia Guiada por Imagen , Pulmón/anomalías , Pulmón/diagnóstico por imagen , Metástasis Linfática/diagnóstico , Neoplasias del Mediastino/diagnóstico por imagen , Mediastinoscopía , Mediastino/patología , Ultrasonografía , Biopsia con Aguja Fina , Femenino , Humanos , Masculino , Persona de Mediana Edad
14.
Biomed Environ Sci ; 30(9): 667-670, 2017 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-29081341

RESUMEN

The study illustrate the inner correlation between global DNA methylation variation and different birth weights. Infant birth weight was used to identify cases and controls. Cord blood and placentas were collected. We performed DNA methylation profiling of bisulphite-converted DNA. We have identified many differentially methylated CpG sites in experimental groups; these sites involved in hundreds of signalings. Among these, more than ten pathways were referred to the glucose and lipid metabolism. Methylation changes in the insulin-signaling pathway (ISP), adipocytokine signaling pathway (ASP) and MAPK signaling pathway are involved in the fetal programming of diabetes..


Asunto(s)
Peso al Nacer , Metilación de ADN , Estudio de Asociación del Genoma Completo , Femenino , Regulación del Desarrollo de la Expresión Génica/fisiología , Humanos , Recién Nacido , Masculino , Tamaño de los Órganos , Placenta/anatomía & histología , Embarazo , Transducción de Señal
15.
J Clin Lab Anal ; 30(4): 351-5, 2016 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-26302683

RESUMEN

BACKGROUND: MicroRNA-27a (miR-27a) is supposed to be an oncogene in various types of cancers, and genetic variation of miR-27a might result in aberrant expression and abnormal second structure of mature-miR-27a, contributing to elevated genetic risk and poor prognosis for colorectal cancer (CRC). METHODS: In order to explore the possible association between rs895819 within miR-27a and CRC in Han Chinese population, we investigated the genotype distributions of rs895819 in 508 CRC cases and 562 healthy check-up controls using TaqMan genotype discrimination system, and analyzed the possible association between them. Odds ratio (OR) and 95% confidential interval (95% CI) were used to assess the strength between allele and genotype of the locus and risk of CRC. RESULTS: In our study, we found that genotype GG of rs895819 was significantly associated with an increased risk for CRC (17.1% vs. 11.6%, adjusted OR = 1.546, 95% CI = 1.070-2.236), and allele A carrier (AA/AG) was significantly associated with a decreased risk for CRC (82.9% vs. 89.4%, adjusted OR = 0.63, 95% CI = 0.446-0.893). In addition, a significant association was observed between genotype GG and larger tumor size (>5 cm; P < 0.001), and allele G was significantly associated with higher pathological stage (TNM-III) (P = 0.008). CONCLUSION: These results indicated that miR-27a might be involved in the development and progression of CRC, genotype GG within rs895819 might be a genetic susceptible factor for CRC. Further multicentral, large sample size, and well-designed epidemiological study as well as functional study are warrant to verify our findings.


Asunto(s)
Pueblo Asiatico/genética , Neoplasias Colorrectales/genética , Etnicidad/genética , Predisposición Genética a la Enfermedad , MicroARNs/genética , Polimorfismo de Nucleótido Simple/genética , Alelos , Estudios de Casos y Controles , Femenino , Humanos , Masculino , Persona de Mediana Edad
16.
Sichuan Da Xue Xue Bao Yi Xue Ban ; 47(1): 90-2, 2016 Jan.
Artículo en Zh | MEDLINE | ID: mdl-27062790

RESUMEN

OBJECTIVE: To investigate the effect of low-intensity anticoagulation therapy for Chinese population with heart valve replacement. METHODS: From January 2011 to March 2012, 3 000 patients with heart valve replacement in Fuwai Hospital and West China Hospital were followed-up for 1 year, the method and intensity of postoperative anticoagulation, as well as the complications were studied and analyzed. RESULTS: The rate of follow-up was 88.57%, and the cumulative follow-up was 1 726.1 patient-years (Pty). The mean oral warfarin dosage was (2.68 ± 6.45) mg/d, and the mean INR values of the patients treated in Fuwai Hospital and West China Hospital were 2.01 ± 1.10 and 186 ± 0.69 respectively (total 54 379 samples). The total rates of anticoagulation complication and mortality were 5.79% Pty and 0.12% Pty respectively, among which the morbidity and mortality of hemorrhage were 3.59% Pty and 0.12% Pty respectively, while the morbidity and mortality of thromboembolism were 2.03% Pty and 0.00% Pty respectively. There are no significant differences of actual anticoagulation intensity (P = 0.28)and the complication rates between the two hospitals . CONCLUSION: The optimal intensity scope (INR) of 1.5-2.5 (mean 1.8-2.0) is safe and efficient for Chinese patients with prosthetic heart valves, and no significant regional difference in the intensity of anticoagulation therapy required.


Asunto(s)
Anticoagulantes/uso terapéutico , Implantación de Prótesis de Válvulas Cardíacas , Pueblo Asiatico , China , Estudios de Seguimiento , Prótesis Valvulares Cardíacas , Hemorragia/mortalidad , Humanos , Periodo Posoperatorio , Tromboembolia/mortalidad , Warfarina/uso terapéutico
17.
BMC Med Genet ; 16: 92, 2015 Oct 08.
Artículo en Inglés | MEDLINE | ID: mdl-26449496

RESUMEN

BACKGROUND: Mitochondrial diabetes is a kind of rare diabetes caused by monogenic mutation in mitochondria. The study aimed to summarize the clinical phenotype profiles in mitochondrial diabetes with m.3243 A>G mitochondrial DNA mutation and to investigate the mechanism in this kind of diabetes by analyzing the relationship among clinical phenotypes and peripheral leukocyte DNA telomere length. METHODS: Fifteen patients with maternally inherited diabetes in five families were confirmed as carrying the m.3243 A>G mitochondrial DNA mutation. One hundred patients with type 2 diabetes and one hundred healthy control subjects were recruited to participate in the study. Sanger sequencing was used to detect the m.3243 A>G mitochondrial DNA mutation. The peak height G/A ratio in the sequence diagram was calculated. Real-time polymerase chain reaction (PCR) was used to measure telomere length. RESULTS: The patients with mitochondrial diabetes all had definite maternally inherited history, normal BMI (19.5 ± 2.36 kg/m(2)), early onset of diabetes (35.0 ± 14.6 years) and deafness. The peak height G/A ratio correlated significantly and negatively with the age at onset of diabetes (≦ 25 years, 61.6 ± 20.17%; 25-45 years, 16.59 ± 8.64%; >45 years, 6.37 ± 0.59%; p = 0.000). Telomere length was significantly shorter among patients with mitochondrial diabetes and type 2 diabetes than in the control group (1.28 ± 0.54 vs. 1.14 ± 0.43 vs. 1.63 ± 0.61; p = 0.000). However, there was no significant difference between patients with mitochondrial diabetes and those with type 2 diabetes. There was no correlation between telomere length and the peak height G/A ratio. CONCLUSION: Deafness with definite maternal inheritance and normal BMI, associated with elevated blood lactic acid and encephalomyopathy, for the most part, suggest the diagnosis of mitochondrial diabetes . The peak height G/A ratio could reflect the spectrum of age at onset of the disease. Telomere length was shorter in patients with mitochondrial diabetes and those with type 2 diabetes, which suggests that the shorter telomere length is likely involved in the pathogenesis of diabetes but is not specific for this kind of diabetes.


Asunto(s)
ADN Mitocondrial/genética , Sordera/genética , Diabetes Mellitus Tipo 2/genética , Estudios de Asociación Genética/métodos , Enfermedades Mitocondriales/genética , Telómero/metabolismo , Adenina/metabolismo , Adolescente , Adulto , Edad de Inicio , Anciano , Sordera/patología , Diabetes Mellitus Tipo 2/patología , Femenino , Guanina/metabolismo , Humanos , Masculino , Persona de Mediana Edad , Enfermedades Mitocondriales/patología , Linaje , Polimorfismo de Nucleótido Simple , Adulto Joven
18.
Yi Chuan ; 37(1): 70-76, 2015 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-25608816

RESUMEN

Peroxisome proliferator-activated receptors (PPARs) are a group of nuclear receptor proteins that regulate gene transcription. PPARs play essential roles in modulating cell differentiation, development, and metabolism (carbohydrate, lipid, protein). Here, we investigated whether PPARγ plays a role in linking maternal malnutrition and aberrant metabolism in the offspring of mice. After feeding dams with high fat (HF) and low protein (LP) diet during pregnancy and lactation, we examined the effects on the offspring at weaning (age of 3-week). The results showed that the LP offspring had lower body weight and length than the control. The HF offspring had heavier body weight and longer body length than LP. The blood glucose levels in HF group were significantly higher at 30 min and 60 min after intraperitoneal glucose administration and the area under curve was also significantly larger than the control. The blood glucose levels in HF group were significantly higher at 30 min than LP. HF group had elevated total cholesterol levels and LP group had decreased total cholesterol levels compared with the control. All results were statistically significant as examined by t-test. More importantly, PPARγ expression levels detected by qRT-PCR were significantly increased in HF and LP groups compared with the control. In conclusion, maternal HF and LP diet during pregnancy and lactation can induce impaired glucose and lipid metabolism in the early life of mouse offspring, where PPARγ may play an important role.


Asunto(s)
Glucosa/metabolismo , Metabolismo de los Lípidos , Desnutrición/metabolismo , Fenómenos Fisiologicos Nutricionales Maternos , PPAR gamma/metabolismo , Efectos Tardíos de la Exposición Prenatal/metabolismo , Alimentación Animal/análisis , Animales , Femenino , Humanos , Masculino , Desnutrición/genética , Ratones , Ratones Endogámicos C57BL , PPAR gamma/genética , Embarazo , Efectos Tardíos de la Exposición Prenatal/genética , Destete
19.
Extremophiles ; 18(2): 195-206, 2014 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-24374718

RESUMEN

A novel haloarchaeal plasmid, pHRDV1 (13,053 bp), was isolated from the haloarchaeal isolate Halorubrum sp. T3. Molecular and bioinformatics analyses showed that this element is a double-stranded circular DNA molecule containing two putative transcripts with opposite directions. The amino acid sequences of six of the nineteen predicted open reading frames were similar to those found in haloarchaeal pleomorphic viruses, such as Halorubrum pleomorphic virus 3 and Halogeometricum pleomorphic virus 1. There was also a strong conservation in gene order between the plasmid and these viruses. All three conserved viral proteins (VPs), which are characteristic of haloarchaeal pleomorphic viruses VP3, VP4 and VP8, were found in pHRDV1. Furthermore, a typical repressor-operator system similar to haloarchaeal myovirus φCh1, was found on the genome of pHRDV1. However, no viral particles were detected in the supernatants of Halorubrum sp. T3, either in the presence or absence of mitomycin C. These results imply that plasmid pHRDV1 is a distinctive virus-like mobile genetic element that harbors some unique properties that make it different from all of the known haloarchaeal plasmids or viruses.


Asunto(s)
Virus de Archaea/genética , Halorubrum/genética , Secuencias Repetitivas Esparcidas , Secuencia de Aminoácidos , Secuencia de Bases , ADN de Archaea/genética , Genes Arqueales , Datos de Secuencia Molecular , Sistemas de Lectura Abierta , Plásmidos
20.
Zhongguo Zhong Yao Za Zhi ; 39(3): 544-7, 2014 Feb.
Artículo en Zh | MEDLINE | ID: mdl-24946565

RESUMEN

To explore the effect of tripterygium glycosides on the level of peripheral blood cell factors of Graves ophthalmopathy (GO). In the study, 64 patients of GO in moderate-severe acute stage were selected, and randomly divided into the treatment group (32 cases) and the control group (32 cases). Both of the two groups were provided with basic treatment. The control group was added with prednisone(0. 75 mg kg-1 d-1 ), which gradually reduced (by 5-10 mg week-1 )to the minimum dose of 5 mg d-1. The treatment group was treated with 20 mg tripterygium glycosides, three times a day. One therapy course is three months. The levels of peripheral blood cells(TNF-alpha , IL-2, IL-10, IFN-gamma)of the two groups before and after the treatment and the clinical efficacy were observed. The study indicated that, before the treatment, TNF-alpha, IL-2, IFN-gamma in both groups were significantly higher than that in the health group, but with IL-10 notably lower than the healthy group. After the treatment, TNF-a, IL-2, IFN-gamma in the treatment group significantly decreased, but with IL-10 significantly increasing (P <0. 01). After the treatment, the two groups showed significant difference (P <0. 01). The total clinical efficacy in the treatment group was 88. 10% , and that in the control group was 57. 14% (P <0. 01). After the treatment, the two groups showed significant changes in the exophthalmos degree (P < 0. 01). The results showed that the level of peripheral blood cells (TNF-alpha,IL-2, IL-10, IFN-gamma)of GO patients was positively correlated with the severity of ocular disease. The combined therapy of tripterygium glycosides and methimazole show such advantages as low side effect and high clin-


Asunto(s)
Citocinas/sangre , Glicósidos/farmacología , Oftalmopatía de Graves/sangre , Oftalmopatía de Graves/tratamiento farmacológico , Tripterygium/química , Adulto , Femenino , Glicósidos/uso terapéutico , Humanos , Masculino
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