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The BRCA2 tumor suppressor protects genome integrity by promoting homologous recombination-based repair of DNA breaks, stability of stalled DNA replication forks and DNA damage-induced cell cycle checkpoints. BRCA2 deficient cells display the radio-resistant DNA synthesis (RDS) phenotype, however the mechanism has remained elusive. Here we show that cells without BRCA2 are unable to sufficiently restrain DNA replication fork progression after DNA damage, and the underrestrained fork progression is due primarily to Primase-Polymerase (PRIMPOL)-mediated repriming of DNA synthesis downstream of lesions, leaving behind single-stranded DNA gaps. Moreover, we find that BRCA2 associates with the essential DNA replication factor MCM10 and this association suppresses PRIMPOL-mediated repriming and ssDNA gap formation, while having no impact on the stability of stalled replication forks. Our findings establish an important function for BRCA2, provide insights into replication fork control during the DNA damage response, and may have implications in tumor suppression and therapy response.

The time evolution of the polarization of a rubidium atom spin ensemble driven by a resonant radio-frequency (RF) magnetic field is analyzed based on the rate equation. A simple optical pumping experimental system is constructed and the time response of the rubidium atomic ensemble is demonstrated by recording the transmitted intensity of pumping light. In the steady-state response, the polarization difference between the optical pumping steady state and the magnetic resonance steady state depends on the optical pumping power and RF magnetic intensity. We can obtain the optimal power value corresponding to the maximum polarization difference. In terms of transient response, where the intensity of RF magnetic field is too weak to observe Rabi oscillations, two decay processes between magnetic resonance and optical pumping steady states are monitored. The decay time from magnetic resonance steady state to optical pumping steady state depends on the optical pumping rate and the spin relaxation rate. The decay time from optical pumping steady state to magnetic resonance steady state depends on the optical pumping rate, the RF driving rate, and the spin relaxation rate. The scale factor of pumping rate to pumping power is obtained, in addition to that of RF driving rate to the RF magnetic field. It can provide an intuitive understanding of the spin dynamic evolution of the polarized atomic ensemble. [ABSTRACT FROM AUTHOR]

A research has been conducted to develop an 8MW electron cyclotron resonance heating and current drive (ECRH/ECCD) system on HL-2M tokamak. The ECRH system compromise eight 1MW gyrotrons, eight evacuated transmission lines and three launchers. The main purpose of the ECRH system was to suppress the neo-classical tearing modes and control the plasma profile. This paper presents an overview of the design and studies performed in this framework. Some primary test results of the critical components have been released in this paper, e.g. polarizers, power monitor and fast steering launchers.

Background:Calreticulin (CALR) exon 9 frameshift mutations have recently been identified in 30–40% of patients with essential thrombocythemia (ET) and primary myelofibrosis (PMF) without JAK2 or MPL mutations. We aimed to develop a qPCR assay to screen type I and II mutations of CALR. Methods: Three different fluorescent-labeled hydrolysis probes and one pair of primers in a closed-tube system were developed to detect CALR type I and II mutations and distinguish them from wild-type. The sensitivity and specificity were validated using TA-cloning plasmids containing CALR wild-type and type I and II mutants, respectively. Fifty-nine ET and PMF specimens were screened by TaqMan qPCR and sequenced by Sanger sequencing. For intra-assay validation, 20 replicates of the assay were performed with each sample. For inter-assay validation, four replications of each sample were carried out and repeated continuously for 5 days. Results: We found that triplex probe-based TaqMan qPCR was reliable in detecting CALR type I and II mutants within DNA that was diluted to 1% of total DNA with the wild-type DNA as background. In 59 patient specimens, six of the observed mutations of CALR were type I and five were type II. Genotyping results obtained from TaqMan qPCR were 100% concordant with Sanger sequencing. The intra- and inter-assay CVs of TaqMan qPCR were less than 3%, respectively. Conclusions: Triplex probe-based TaqMan qPCR is an accurate and sensitive method for screening ET or PMF patients with type I and II mutations in CALR. [ABSTRACT FROM AUTHOR]

Ephrin-A2, a member of the Eph/ephrin family, is associated with tumorigenesis and tumor progression. This study aimed to assess the diagnostic and prognostic value of both serum and tissue levels of Ephrin-A2 in prostate cancer (PCa) management. One hundred and forty-five frozen prostate tissues, 55 paraffin-embedded prostate tissues, 88 serum samples, and seven prostate cell lines (RWPE-1, LNCaP, LNCaP-LN3, PC-3, PC-3M, PC-3M-LN4, and DU145) were examined via quantitative reverse transcription-PCR (qRT-PCR), immunohistochemistry, enzyme-linked immunosorbent assay, and western blotting. Induced Ephrin-A2 messenger RNA (mRNA) or protein expression was detected in 8.6 % (5/58) benign prostatic hyperplasia (BPH), 59.8 % (52/87) PCa, and five prostate cancer cell lines. Ephrin-A2 immunostaining was present in 6.7 % (1/15) patients with BPHs and 62.5 % (25/40) clinically localized PCa. Accordingly, serum Ephrin-A2 was significantly higher in PCa patients compared to those in the BPH patients and controls ( P < 0.001). The expression of Ephrin-A2 was higher in tumor patients with an elevated Gleason score or T3-T4 staging. Ephrin-A2 expression was correlated with Ki-67 expression in PCa patients, both at the gene scale and protein level. Our data indicate that Ephrin-A2 is a potential diagnostic and prognostic biomarker and a promising molecular therapeutic target to attenuate prostate cancer progression. [ABSTRACT FROM AUTHOR]

Background: Extramammary Paget's disease (EMPD) is a rare skin tumor. Hypermethylation in the MSH 2 promoter resulting in the downregulation of its protein expression shows a high detection rate in EMPD tumor tissue, which indicates that the methylation of MSH 2 may play an important role in the pathogenesis of EMPD., Objective: This study aims to establish a rapid analysis strategy based on the methylation-sensitive high-resolution melting curve (MS-HRM) to detect the methylation level of the MSH 2 promoter., Methods: With the use of universal methylated human DNA products, we established the MS-HRM standard curve to quantitatively detect the methylation level of the MSH 2 promoter. Then, all 57 EMPD tumor DNA samples were analyzed. Pyrosequencing assay was also carried out to test the accuracy and efficacy of MS-HRM. Besides, a total of 54 human normal and other cancerous tissues were included in this study to test the reliability and versatility of the MS-HRM standard curve., Results: In this study, by using the established MS-HRM, we found that 96.5% (55/57) EMPD tumor samples had varying methylation levels in the MSH 2 promoter ranging from 0% to 30%. Then, the methylation data were compared to the results obtained from pyrosequencing, which showed a high correlation between these two techniques by Pearson's correlation ( r  = 0.9425) and Bland-Altman plots (mean difference = -0.1069) indicating that the methylation levels analyzed by MS-HRM were consistent with DNA pyrosequencing. Furthermore, in 23 normal and 31 other cancerous tissue samples, there were two colorectal cancer (CRC) tissues that tested MSH 2 methylation positive (1% and 5%) which confirmed that our established MS-HRM can be widely applied to various types of samples., Conclusion: MS-HRM standard curve can be used for the detection of the methylation level of MSH 2 in EMPD tumor samples and other cancerous tissues potentially, which presents a promising candidate as a quantitative assay to analyze MSH 2 promoter methylation in routine pathological procedure., Competing Interests: The authors declare no conflicts of interest., (Copyright © 2021 Liu Dong et al.)

Skin, as the major organ of a human body, is constantly exposed to PM 2.5 stimulation, which may exert specific toxic influences on the physiology of skin. This study aims to investigate the effect of PM 2.5 on the formation of inflammasomes in skin cells and to explore the potential mechanism linking PM 2.5 and skin inflammation. Changes in mRNA and protein levels of inflammasome-related genes were detected by real-time PCR and western blot in human immortalized epidermal cells (HaCaT) treated with PM 2.5 at multiple concentrations for 24 hours. The expression of NLRP1 was increased significantly both in mRNA and protein levels after PM 2.5 exposure while the elevated secretory protein level of IL-1β in cell culture was detected by ELISA, which is one of the main downstream factors of NLRP1. In addition, the upregulation of NLRP1 and IL-1β could be reversed by NF-κB inhibitor indicating that PM 2.5 may promote NLRP1 expression through activating NF-κB pathway. Furthermore, high ROS level was also found in cells treated with PM 2.5 and inhibition of ROS could also reverse NK-κB production stimulated by PM 2.5 that means ROS is involved in this skin inflammation process., Competing Interests: Competing Interests: The authors have declared that no competing interest exists., (© The author(s).)