RESUMEN
Rapid pathogen detection is critical for prompt treatment, interrupting transmission routes, and decreasing morbidity and mortality. The V-type CRISPR system had been used for rapid pathogen detection. However, whether single-stranded DNA in CRISPR system can cause false positives remains undetermined. Herein, we show that high molar concentration of Cas12a effector tolerated more mismatches on ssDNA and activated its trans-cleavage activity at six base matches. Reducing Cas12a and crRNA molar concentration increased the minimal base-match number required for Cas12a ssDNA activation to 11, which reducing nonspecific activation. We then established a Cas12a-based M tuberculosis detection system with a primer having an 8 bp overlap with crRNA. This system did not exhibit primer-induced false positives, and minimum detection copy reached 1 copy/uL (inputting 1-µL sample) in standard strains. The Cas12a-based M tuberculosis detection system showed 80.0% sensitivity and 100.0% specificity in verification using clinical specimens, compared with Xpert MTB/RIF, which showed 72.0% sensitivity and 90.9% specificity. All these results prove that appropriate concentration of cas12a effector can effectively perform nucleic acid detection.
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Proteínas Bacterianas/química , Proteínas Asociadas a CRISPR/química , Sistemas CRISPR-Cas , ADN de Cadena Simple/química , Endodesoxirribonucleasas/química , Mycobacterium tuberculosis/química , HumanosRESUMEN
AIM: Gestational diabetes mellitus (GDM) is the most common complication in pregnancy. This study aimed to investigate the potential mechanism and effects of long-noncoding RNA maternally expressed 8 (lncRNA-MEG8) in GDM. METHODS: Targeted interactions involving lncRNA-MEG8 and miR-296-3p were initially predicted using starBase software and then confirmed using dual-luciferase reporter gene analysis. The expression levels of lncRNA-MEG8 and miR-296-3p in peripheral blood samples from patients with GDM were measured using reverse transcription-quantitative polymerase chain reaction. Enzyme-linked immunosorbent assay was used to evaluate the overall levels of insulin and insulin secretion. Additionally, MTT and flow cytometric methods were used to detect cell viability and apoptosis. Cell apoptosis-associated proteins were determined by western blotting. RESULTS: Our results indicated that lncRNA-MEG8 is a potential target of miR-296-3p. lncRNA-MEG8 level was higher, whereas that of miR-296-3p was lower in patients with GDM than in healthy individuals. LncRNA-MEG8-siRNA promoted insulin content and secretion. Furthermore, MEG8-siRNA increased cell viability and decreased apoptosis. However, these changes were reversed by an miR-296-3p inhibitor. Moreover, a miR-296-3p mimic had the same effect on INS-1 cells as MEG8-siRNA, as evidenced by enhanced insulin secretion, cell viability, and reduced apoptosis. CONCLUSION: LncRNA-MEG8-siRNA promotes pancreatic ß-cell function by upregulating miR-296-3p.
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Diabetes Gestacional , Insulinas , MicroARNs , ARN Largo no Codificante , Femenino , Embarazo , Humanos , ARN Largo no Codificante/genética , ARN Largo no Codificante/metabolismo , Diabetes Gestacional/genética , MicroARNs/genética , MicroARNs/metabolismo , ARN Interferente Pequeño/farmacología , Proliferación Celular , Apoptosis , Insulinas/farmacologíaRESUMEN
BACKGROUND: The preferred type of postremission therapy (PRT) for intermediate-risk acute myeloid leukemia (AML) in first complete remission (CR1) is a subject of continued debate. Although allogeneic stem cell transplantation (alloSCT) is regarded as a curative strategy for AML, the efficacy of autologous stem cell transplantation (autoSCT) for patients without a matched sibling donor (MSD) has remained controversial. METHODS: To compare survival outcomes after alloSCT versus autoSCT for patients with intermediate-risk AML in CR1, we performed a meta-analysis of 11 clinical studies. The outcomes included relapse-free survival (RFS), overall survival (OS), relapse rate (RR), and treatment-related mortality (TRM). RESULTS: Compared with autoSCT, alloSCT showed better RFS, OS, and RR benefits, but higher TRM. Subgroup analysis based on donor category (MSD and matched unrelated donor [MUD]) of alloSCT showed alloSCT from MSD rather than from MUD had better OS benefits compared to autoSCT. For fms-like tyrosine kinase 3 internal tandem duplications (FLT3-ITD) wild-type patients, alloSCT and autoSCT had comparable RFS and OS outcomes. CONCLUSION: Our results suggest that, in the absence of an available MSD, autoSCT remains a viable PRT alternative for intermediate-risk AML in CR1, especially for FLT3-ITD wild-type patients.
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Leucemia Mieloide Aguda/mortalidad , Leucemia Mieloide Aguda/terapia , Hermanos , Trasplante de Células Madre , Donante no Emparentado , Aloinjertos , Autoinjertos , Supervivencia sin Enfermedad , Femenino , Humanos , Leucemia Mieloide Aguda/genética , Masculino , Tasa de Supervivencia , Tirosina Quinasa 3 Similar a fms/genéticaRESUMEN
Beauveria bassiana and Metarhizium anisopliae are two of the most important and widely used entomopathogenic fungi (EPFs) to control insect pests. Recent studies have revealed their function in promoting plant growth after artificial inoculation. To better assess fungal colonization and growth-promoting effects of B. bassiana and M. anisopliae on crops, maize Zea mays seedlings were treated separately with 13 B. bassiana and 73 M. anisopliae as rhizosphere fungi in a hydroponic cultural system. Plant growth indexes, including plant height, root length, fresh weight, etc., were traced recorded for 35 days to prove the growth promoting efficiency of the EPFs inoculation. Fungal recovery rate (FRR) verified that both B. bassiana and M. anisopliae could endophytically colonize in maize tissues. The recovery rates of B. bassiana in stems and leaves were 100% on the 7th day, but dropped to 11.1% in the stems and 22.2% in the leaves on the 28th day. Meanwhile, B. bassiana was not detected in the roots until the 28th day, reaching a recovery rate of 33.3%. M. anisopliae strains were isolated from the plant roots, stems and leaves throughout the tracing period with high recovery rates. The systematical colonization of B. bassiana and M. anisopliae in different tissues were further corroborated by PCR amplification of fungus-specified DNA band, which showed a higher detection sensitivity of 100% positive reaction. Fungal density comparing to the initial value in the hydroponic solution, dropped to be well below 1% on the 21st day. Thus, the two selected entomopathogenic fungal strains successfully established endophytic colonization rather than rhizospheric colonization in maize, and significantly promoted its growth in a hydroponic cultural system. Entomopathogenic fungi have great application potential in eco-agricultural fields including biopesticides and biofertilizers.
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Beauveria , Metarhizium , Agentes de Control Biológico , ADN , Control Biológico de Vectores , Zea maysRESUMEN
OBJECTIVE: This study was to investigate the association of Bisphenol A and unexplained recurrent spontaneous abortion. METHODS: A hospital-based 1:1 matched case-control study was conducted. Sixty patients with unexplained recurrent abortion were included. Each case was matched with one normal control by age (± 2 years), living district and the same gestational age. The levels of Bisphenol A in urine for 60 cases and 60 controls were detected using high performance liquid chromatography after fluorescent derivatization. The levels of urinary Bisphenol A in case was compared with that in control in education levels, occupation, smoking history. Data was analyzed by means of Wilcoxon-test, Student-Newman-Keuls after rank transform, univariate and multivariate conditional Logistic regression analysis. The software used was SAS 9.1.3. RESULTS: The values of urinary Bisphenol A in cases and controls were (0.10 ± 0.21) µg/ml, (0.03 ± 0.08) µg/ml, respectively. The level of urinary Bisphenol A in cases was significantly higher than that in controls (Z = 3.988, P < 0.0001). The urinary Bisphenol A levels in cases were significant higher than those in controls from senior middle school (the levels were 0.10, 0.06 µg/ml respectively, Z = 1.996, P = 0.0459), college (the levels were 0.14, 0.03 µg/ml respectively, Z = 2.586, P = 0.0097), workers or farmers (the levels were 0.08, 0.03 µg/ml respectively, Z = 2.265, P = 0.0235), businessmen (the levels were 0.10, 0.03 µg/ml respectively, Z = 2.544, P = 0.0109), and no passive smokers (the levels were 0.09, 0.03 µg/ml respectively, Z = 3.767, P = 0.0002). After adjustment by age, body mass index, marital status during pregnancy and other factors, compared to Bisphenol A below 0.06 µg/ml, the adjusted OR was 4.03 (1.67 - 9.74) for Bisphenol A levels between 0.06 µg/ml and 0.20 µg/ml, and was 5.46 (1.95 - 15.27) for Bisphenol A over 0.20 µg/ml. The risk of unexplained recurrent spontaneous abortion increased progressively with the growth of urinary Bisphenol A levels (χ(2) = 13.042, trend test P = 0.0003). There were significant differences on Bisphenol A among controls, two abortions, and three or more abortions (the levels were 0.03 µg/ml, 0.09 µg/ml, 0.21 µg/ml respectively, F = 9.04, P = 0.0002). CONCLUSION: Exposure to Bisphenol A may be associated with unexplained recurrent spontaneous abortion.
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Aborto Habitual/etiología , Aborto Espontáneo/etiología , Exposición Materna/efectos adversos , Fenoles/orina , Adulto , Compuestos de Bencidrilo , Estudios de Casos y Controles , Causalidad , Femenino , Humanos , Embarazo , Adulto JovenRESUMEN
BACKGROUND: Evidence about the association between Bisphenol A (BPA) and the risk of recurrent miscarriage (RM) in human being is still limited. OBJECTIVE: We evaluated the association of urinary BPA concentrations with RM in human being. METHODS: A hospital-based 1:2 matched case-control study on RM was carried out in Suzhou and Kunshan in Jiangsu Province in China between August 2008 and November 2011. Total urinary BPA concentrations in 264 eligible urine samples (102 RM patients and 162 controls) were measured using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The Wilcoxon test and conditional logistic regression were used to estimate the differences between the groups and odds ratios (OR) with 95% confidence intervals (CI), respectively. RESULTS: The median ± IQR (interquartile range) (P75-P25) values of non-creatinine-adjusted total urinary BPA levels in the RM patients and the controls were 1.66 ± 3.69 ng/ml and 0.58 ± 1.07 ng/ml, respectively (0.98 ± 2.67 µg/g Cr (creatinine) and 0.40 ± 0.77 µg/g Cr. The adjusted BPA level was significantly higher in the RM patients than in the controls (Wilcoxon test, Z = 4.476, P < 0.001). Higher level of urinary BPA was significantly associated with an increased risk of RM (P-trend < 0.001). Compared to the groups with urinary BPA levels less than 0.16 µg/g Cr, the women with levels of 0.40-0.93 µg/g Cr and 0.93 µg/g Cr or above had a significantly higher risk of RM (OR = 3.91, 95%CI: 1.23-12.45 and OR = 9.34, 95%CI: 3.06-28.44) that persisted after adjusting for confounding factors. The time from recently RM date to recruitment does not significantly influence the urinary BPA level (P = 0.090). CONCLUSION: Exposure to BPA may be associated with RM risk.