Real-time PCR is the most utilized nucleic acid testing tool in clinical settings. However, the number of targets detectable per reaction are restricted by current modes. Here, we describe a single-step, multiplex approach capable of detecting dozens of targets per reaction in a real-time PCR thermal cycler. The approach, termed MeltArray, utilizes the 5′-flap endonuclease activity of Taq DNA polymerase to cleave a mediator probe into a mediator primer that can bind to a molecular beacon reporter, which allows for the extension of multiple mediator primers to produce a series of fluorescent hybrids of different melting temperatures unique to each target. Using multiple molecular beacon reporters labeled with different fluorophores, the overall number of targets is equal to the number of the reporters multiplied by that of mediator primers per reporter. The use of MeltArray was explored in various scenarios, including in a 20-plex assay that detects human Y chromosome microdeletions, a 62-plex assay that determines Escherichia coli serovars, a 24-plex assay that simultaneously identifies and quantitates respiratory pathogens, and a minisequencing assay that identifies KRAS mutations, and all of these different assays were validated with clinical samples. MeltArray approach should find widespread use in clinical settings owing to its combined merits of multiplicity, versatility, simplicity, and accessibility.
Glioblastoma multiforme (GBM) is the most common, malignant and high-grade brain tumour. 1,2 The WHO classification system divides glioma into 4 subtypes and GBM as grade 4 glial tumour has the worst prognosis. 3 The intra-and intertumoral genetic and epigenetic heterogeneity observed in GBM highlights the complexity of cancer. 4 The median survival of GBM patients is around 12-15 months, even with
Background: Recurrent implantation failure (RIF) is a critical problem for assisted reproduction technology. High-quality embryos and the synchronization endometrium both have great significance. How to get the optimal endometrial receptivity is a challenge for implantation and pregnancy of infertile patients with RIF. The objective of this study is to investigate personalized protocol of frozen-thawed embryo transfer (FET) cycles, and its effect on clinical outcomes in patients with RIF.Methods: We chose 91 RIF patients from January 2017 to June 2019 in the Reproductive Medicine Center of the First Hospital of Lanzhou University. A total of 100 FET cycles were undertaken with a gonadotropinreleasing hormone agonist (GnRH-agonist) protocol combined with hormone replacement therapy (HRT) for endometrial preparation. The patients were divided into two groups: the routine group (cleavage embryo transferred at day 3 after luteal support) included 48 cycles; the personalized group included 52 cycles with delayed endometrial trigger and luteal support (the time of embryo transfer depended on the level of serum hormone and endometrial thickness).
Results:The data showed the personalized group had longer time for endometrial preparation. On the day of embryo transfer, serum progesterone (P) and the E 2 /P ratio was significantly different compared with the routine group (P<0.05). The clinical pregnancy rate in the routine group was 35.42% (17/48) and 59.62% (31/52) in the personalized group (P<0.05). The abortion rate was not significantly different.Conclusions: For women with RIF, personalized timing for transfer of FET resulted in a higher clinical pregnancy rate compared with routine protocol.
ObjectiveTo evaluate the utility of apparent diffusion coefficient (ADC) values for differentiating breast tumors.MethodsThe medical records of 17 patients with phyllodes tumor [PT; circular regions of interest (ROI-cs) n = 171], 74 patients with fibroadenomas (FAs; ROI-cs, n = 94), and 57 patients with breast cancers (BCs; ROI-cs, n = 104) confirmed by surgical pathology were retrospectively reviewed.ResultsThere were significant differences between PTs, FAs, and BCs in ADCmean, ADCmax, and ADCmin values. The cutoff ADCmean for differentiating PTs from FAs was 1.435 × 10−3 mm2/s, PTs from BCs was 1.100 × 10−3 mm2/s, and FAs from BCs was 0.925 × 10−3 mm2/s. There were significant differences between benign PTs, borderline PTs, and malignant PTs in ADCmean, ADCmax, and ADCmin values. The cutoff ADCmean for differentiating benign PTs from borderline PTs was 1.215 × 10−3 mm2/s, and borderline PTs from malignant PTs was 1.665 × 10−3 mm2/s.ConclusionDWI provides quantitative information that can help distinguish breast tumors.
Acute myeloid leukemia (AML) is afflicted by a high‐mortality rate and few treatment options. The lack of specific surface antigens severely hampers the development of targeted therapeutics and cell therapy. Here, it is shown that exogenous all‐trans retinoic acid (ATRA) mediates selective and transient CD38 upregulation on leukemia cells by up to 20‐fold, which enables high‐efficiency targeted nanochemotherapy of leukemia with daratumumab antibody‐directed polymersomal vincristine sulfate (DPV). Strikingly, treatment of two CD38‐low expressing AML orthotopic models with ATRA and DPV portfolio strategies effectively eliminates circulating leukemia cells and leukemia invasion into bone marrow and organs, leading to exceptional survival benefits with 20–40% of mice becoming leukemia‐free. The combination of exogenous CD38 upregulation and antibody‐directed nanotherapeutics provides a unique and powerful targeted therapy for leukemia.
This research aimed to prepare superabsorbent resin from agro-mining waste, which is important for sustainable development and practical application in agriculture and forestry. Corn stover (CS), humic acid (HA), acrylic acid (AA), and acrylamide (AM) were used as raw materials to produce CS-HA-based superabsorbent resin (p-CS-HA-PAA) by plasma modification, and the optimum modification conditions were obtained. The superabsorbent resin was characterized by Fourier transform infrared spectroscopy, scanning electron microscopy, and contact angle analysis. The adsorption characteristics of the superabsorbent resin were analyzed by kinetic model fitting, and the performances were tested. The results showed that the water absorption capacity could reach 1106.9 g/g under the following optimal process conditions: a discharge time of 180 s, a discharge power of 40 W, and a discharge pressure of 20 Pa. The characterization confirmed that plasma modification could make the surface of the superabsorbent resin rougher, produce more and larger pores, and introduce more hydrophilic groups. The p-CS-HA-PAA conformed to the pseudo-second-order kinetic equation and non-Fickian intradiffusion process in aqueous solution. The chemical resistance, repeated liquid absorption, and water retention properties of the modified resin were better than those of the unmodified resin, which provided a viable way for industrial and agricultural applications.
A new method for the direct determination of acrylamide residues in poly-acrylamide (PAM) by ultra-performance liquid chromatography (UPLC)-quadrupole electrostatic field orbitrap high-resolution mass spectrometry with the Hypercarb column was developed. With being diluted by water, the PAM sample was dissolved well by high-speed stirring. After filtering through a 0.22 μm aqueous phase microporous membrane, the sample was analyzed by high-resolution mass spectrometry directly. Chromatographic analysis was carried out using a Hypercarb column, distilled water and methanol applied as the mobile phases. In addition, the column temperature was strictly set at 20℃. The sample was determined in the mode of parallel reaction monitoring (PRM). Accurate mass-to-charge ratio extraction of the target compound ion and its fragment ion were utilized with elemental composition analysis and isotope distribution of the fragment ion to further improve the accuracy of qualitative and quantitative analysis. Meanwhile, the fragmentation pattern of acrylamide was acquired. The calibration curve showed a good linearity in the range of 2-50 μg/L with the correlation coefficient of 0.9998. The limit of detection (LOD) was 1.5 μg/kg, satisfying the detection demands. The recoveries of the acrylamide were in the range of 101.3%-107.1%. The relative standard deviations (RSD) were 3.1%-4.1%. The applications indicated that the mass percentages of AM in PAM were in the range of 0-0.43%. The proposed method is easy, fast, sensitive and suitable for the determination and confirmation of acrylamide residues in PAM.
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