MiRNA-let-7b smanjuje proliferacijsku aktivnost i razvoj folikularnih stanica putem ciljnog gena MAP3K1

To date, it has not yet been determined if the apoptosis of follicular granulosa cells (FGCs) is mediated by miR- let-7b via MAP3K1. In the present study, FGCs were transfected with a miR-let-7b mimic at different doses (0, 40, 60, 80, 100 and 120 μM), and were allocated into the control group (CG), and MIM-1, MIM-2, MIM-3, MIM-4 and MIM- 5, respectively. Expression levels of miR-let-7b and mitogen-activated protein kinase kinase kinase 1 (MAP3K1) mRNAs and proteins were determined using RT-PCR and Western blots. Luciferase report assay was applied to verify the targeting relationship between miR-let-7b and MAP3K1. The results revealed that the proliferation activity of FGCs in the MIM-4 group was significantly lower than that of the CG and MIM-1 groups (P<0.05). The MiR-let-7b mimic obviously reduced expression levels of miR-let-7b of the FGCs. The largest reduction was found in MIM-4. Levels of MAP3K1 mRNAs and proteins in the MIM-3 and MIM-4 groups were lower than that of the CG (P<0.05 or P<0.01). Co-transfection of let-7b mimic significantly inhibited luciferase activity (P<0.05) as compared with the CG. In conclusion, miR-let-7b may obviously depress the cell viability and accelerate apoptosis of ovine FGCs. Higher doses of miR-let-7b mimic (80 μM and 100 μM) could significantly depress expressions of miR-let-7b mRNAs, MAP3K1 mRNAs and protein in ovine FGCs. MiR-let-7b promoted FGCs apoptosis by inhibiting the MAP3K1 gene.Do danas nije definirano je li apoptoza folikularnih granuloza-stanica (FGCs) posredovana prekursorom miR-let- 7b putem gena MAP3K1. U ovom je istraživanju FGC transfeciran miR-let-7b imitatorom u različitim dozama (0, 40, 60, 80, 100 i 120 μM) i potom razvrstan u kontrolnu skupinu (CG), te u skupine označene kao MIM-1, MIM-2, MIM-3, MIM-4 i MIM-5. Razine ekspresije mRNA i proteina miR-let-7b i mitogenom-aktivirane proteinske kinase kinase kinase1 (MAP3K1) određene su uporabom RT-PCR-a i Western blot-a. Test luciferaze je primijenjen kako bi se potvrdio ciljni odnos između miR-let-7b i MAP3K1. Rezultati su pokazali da je proliferacijska aktivnost FGC-a u skupini MIM-4 bila znakovito manja nego u kontrolnoj skupini i MIM-1 skupini (P<0,05) što ide u prilog pretpostavci da miR-let-7b imitator može u FGCs smanjiti razinu ekpresije miR-let-7b. Najveće smanjenje utvrđeno je u skupini MIM-4. Razine MAP3K1 mRNAs i proteina u skupinama MIM-3 i MIM-4 bile su niže nego u kontrolnoj skupini (P<0,05; P<0,01). Transfekcija let-7b imitatorom znakovito je inhibirala aktivnost luciferaze (P<0,05) u usporedbi s kontrolnom skupinom. Zaključno, miR-let-7b može smanjiti opstojnost stanice i ubrzati apoptozu ovčjih FGC-a. Veće doze miR-let-7b mimic (80 μM i 100 μM) mogu znakovito smanjiti ekspresiju miR-let-7b mRNAs, MAP3K1 mRNAs i proteina u ovčjih FGC-a. MiR-let-7b putem inhibirajućeg gena MAP3K1 doprinosi apoptozi folikularnih granuloza-stanica (FGCs)

Application of plasma metagenomic next-generation sequencing improves prognosis in hematology patients with neutropenia or hematopoietic stem cell transplantation for infection

IntroductionMetagenomic next-generation sequencing (mNGS) is a novel technique for detecting pathogens. This retrospective study evaluated the diagnostic value of mNGS using plasma for infections in hematology patients and its impact on clinical treatment and prognosis in different subgroups of hematology patients.MethodsA total of 153 hematology patients with suspected infection who underwent mNGS using plasma were enrolled in the study. Their clinical histories, conventional microbiological test (CMT) results, mNGS results, treatment and prognosis were retrospectively analyzed.ResultsIn 153 plasma samples, mNGS yielded a higher positivity rate than CMT (total: 88.24% vs. 40.52%, P&lt;0.001; bacteria: 35.95% vs. 21.57%, P &lt; 0.01; virus: 69.93% vs. 21.57%, P&lt;0.001; fungi: 20.26% vs. 7.84%, P&lt;0.01). mNGS had a higher positivity rate for bacteria and fungi in the neutropenia group than in the non-neutropenia group (bacteria: 48.61% vs. 24.69%, P&lt;0.01; fungi: 27.78% vs. 13.58%, P&lt;0.05). mNGS demonstrated a greater advantage in the group of patients with hematopoietic stem cell transplantation (HSCT). Both the 3-day and 7-day efficacy rates in the HSCT group were higher than those in the non-HSCT group (3-day: 82.22% vs. 58.65%, P &lt; 0.01; 7-day: 88.89% vs. 67.31%, P &lt; 0.01), and the 28-day mortality rate was lower in the HSCT group than in the non-HSCT group (6.67% vs. 38.89%, P &lt; 0.000). The neutropenia group achieved similar efficacy and mortality rates to the non-neutropenia group (7-day efficiency rate: 76.39% vs. 71.43%, P &gt; 0.05; mortality rate: 29.17% vs. 29.63%, P &gt; 0.05) with more aggressive antibiotic adjustments (45.83% vs. 22.22%, P &lt; 0.01).ConclusionmNGS can detect more microorganisms with higher positive rates, especially in patients with neutropenia. mNGS had better clinical value in patients with hematopoietic stem cell transplantation (HSCT) or neutropenia, which had a positive effect on treatment and prognosis

Scalable deterministic integration of two quantum dots into an on-chip quantum circuit

Integrated quantum photonic circuits (IQPCs) with deterministically integrated quantum emitters are critical elements for scalable quantum information applications and have attracted significant attention in recent years. However, scaling up them towards fully functional photonic circuits with multiple deterministically integrated quantum emitters to generate photonic input states remains a great challenge. In this work, we report on a monolithic prototype IQPC consisting of two pre-selected quantum dots deterministically integrated into nanobeam cavities at the input ports of a 2x2 multimode interference beam-splitter. The on-chip beam splitter exhibits a splitting ratio of nearly 50/50 and the integrated quantum emitters have high single-photon purity, enabling on-chip HBT experiments, depicting deterministic scalability. Overall, this marks a cornerstone toward scalable and fully-functional IQPCs

Comprehensive Analysis of the Relationship Between RAS and RAF Mutations and MSI Status of Colorectal Cancer in Northeastern China

Background/Aims: Colorectal cancer (CRC) is mainly caused by chromosomal instability (CIN) and microsatellite instability (MSI). The RAS and RAF genes are essential components of the CIN pathway, and several studies have found that RAS and RAF mutations are associated with MSI status in CRC. Here, we examined these three factors in CRC in Northeast China and aimed to reveal new details of the relationship between these mutations and MSI status. Methods: This study involved 290 patients with CRC who had RAS or RAF gene mutation detected using fluorescence-based allele-specific polymerase chain reaction or Sanger sequencing. The majority of the identified patients were found to harbor MSI (MSI status). Accurate molecular detection was carried out using formalin-fixed paraffin-embedded tissue or blood samples. Results: The rates of RAS and RAF mutations were 58.5% and 4.1%, respectively. The prevalence of RAS mutation in CRC was clearly higher and that of RAF mutation was lower in Northeast China compared with previously reported cohorts in other locations. High MSI level (MSI-H status) was more complex, at around 10%. This was consistent with previous data from China. However, compared with data reported from other continents, MSI-H was higher than that of Japan or South Korea in Asia, and lower than that of Europe or the United States. Conclusion: RAS/RAF mutations and MSI status in CRC are closely associated with tumor location and ethnicity. Further studies investigating the relationship between these three factors can help in the development of treatment strategies for patients with CRC

Qishen Yiqi dripping pills for chronic ischaemic heart failure:results of the CACT-IHF randomized clinical trial

10.1002/ehf2.12980ESC Heart Failure763881-389

Robust estimation of bacterial cell count from optical density

Optical density (OD) is widely used to estimate the density of cells in liquid culture, but cannot be compared between instruments without a standardized calibration protocol and is challenging to relate to actual cell count. We address this with an interlaboratory study comparing three simple, low-cost, and highly accessible OD calibration protocols across 244 laboratories, applied to eight strains of constitutive GFP-expressing E. coli. Based on our results, we recommend calibrating OD to estimated cell count using serial dilution of silica microspheres, which produces highly precise calibration (95.5% of residuals &lt;1.2-fold), is easily assessed for quality control, also assesses instrument effective linear range, and can be combined with fluorescence calibration to obtain units of Molecules of Equivalent Fluorescein (MEFL) per cell, allowing direct comparison and data fusion with flow cytometry measurements: in our study, fluorescence per cell measurements showed only a 1.07-fold mean difference between plate reader and flow cytometry data

Omecamtiv mecarbil in chronic heart failure with reduced ejection fraction, GALACTIC‐HF: baseline characteristics and comparison with contemporary clinical trials

Aims: The safety and efficacy of the novel selective cardiac myosin activator, omecamtiv mecarbil, in patients with heart failure with reduced ejection fraction (HFrEF) is tested in the Global Approach to Lowering Adverse Cardiac outcomes Through Improving Contractility in Heart Failure (GALACTIC‐HF) trial. Here we describe the baseline characteristics of participants in GALACTIC‐HF and how these compare with other contemporary trials. Methods and Results: Adults with established HFrEF, New York Heart Association functional class (NYHA) ≥ II, EF ≤35%, elevated natriuretic peptides and either current hospitalization for HF or history of hospitalization/ emergency department visit for HF within a year were randomized to either placebo or omecamtiv mecarbil (pharmacokinetic‐guided dosing: 25, 37.5 or 50 mg bid). 8256 patients [male (79%), non‐white (22%), mean age 65 years] were enrolled with a mean EF 27%, ischemic etiology in 54%, NYHA II 53% and III/IV 47%, and median NT‐proBNP 1971 pg/mL. HF therapies at baseline were among the most effectively employed in contemporary HF trials. GALACTIC‐HF randomized patients representative of recent HF registries and trials with substantial numbers of patients also having characteristics understudied in previous trials including more from North America (n = 1386), enrolled as inpatients (n = 2084), systolic blood pressure &lt; 100 mmHg (n = 1127), estimated glomerular filtration rate &lt; 30 mL/min/1.73 m2 (n = 528), and treated with sacubitril‐valsartan at baseline (n = 1594). Conclusions: GALACTIC‐HF enrolled a well‐treated, high‐risk population from both inpatient and outpatient settings, which will provide a definitive evaluation of the efficacy and safety of this novel therapy, as well as informing its potential future implementation