Supplementary Data from Targeting Squalene Epoxidase Interrupts Homologous Recombination via the ER Stress Response and Promotes Radiotherapy Efficacy

Summary: Increased PD-L1 expression in cancer cells is known to enhance immunosuppression, but the mechanism underlying PD-L1 upregulation is incompletely characterized. We show that PD-L1 expression is upregulated through internal ribosomal entry site (IRES)-mediated translation upon mTORC1 inhibition. We identify an IRES element in the PD-L1 5′-UTR that permits cap-independent translation and promotes continuous production of PD-L1 protein despite effective inhibition of mTORC1. eIF4A is found to be a key PD-L1 IRES-binding protein that enhances PD-L1 IRES activity and protein production in tumor cells treated with mTOR kinase inhibitors (mTORkis). Notably, treatment with mTORkis in vivo elevates PD-L1 levels and reduces the number of tumor-infiltrating lymphocytes in immunogenic tumors, but anti-PD-L1 immunotherapy restores antitumor immunity and enhances the therapeutic efficacy of mTORkis. These findings report a molecular mechanism for regulating PD-L1 expression through bypassing mTORC1-mediated cap-dependent translation and provide a rationale for targeting PD-L1 immune checkpoint to improve mTOR-targeted therapy.

为合理评估水汽资源,利用大气红外探测仪(AIRS)和先进微波探测器(AMSU)联合反演的2006―2015年AIRS L2标准反演产品AIRX2RET V006,研究了安徽淮北地区水汽分布和变化特征,重点分析了该地区整层大气可降水量和若干层累积水汽月均值、季均值的年际变化,以及水汽和温度的关系.研究发现:从2006年到2015年这10年间,淮北地区月平均整层大气可降水量呈现逐年减小的趋势.1000～850、850～500和500～100 hPa层累积水汽和整层大气可降水量四季分布一致,呈现夏季>秋季>春季>冬季.四季整层大气可降水量与三层累积水汽年际变化也呈现出较高的一致性,表现为夏季逐年变化相对较大,呈线性减小趋势;秋季次之,除500～100 hPa层外,在2006至2011年期间呈现逐年线性增大趋势;春、冬两季年际变化相对较小,且无明显的线性关系.

The routing protocol for low-power and lossy networks (RPL), standardized by Internet Engineering Task Force (IETF), is mainly designed to use for Low-power and Lossy Networks (LLNs). To solve the problems of several important routing metrics are not evaluated, the optimal path may contain long single hop links, lack of scientific multi-routing metrics evaluation method and mechanism to balance the parent child number (especially the parent with one hop away from root), this paper proposes an improved RPL algorithm for LLN (I-RPL). First of all, we propose the evaluated routing metrics: child number of parent, candidate parent number, hop count, ETX and energy consumption index. Meanwhile, we improve the path ETX calculation method to avoid selecting optimal path containing long single hop links. Then we design a novel lexical method to synthetically evaluate candidate parents. Meanwhile, based on the evaluation results of candidate parents, we design a novel objective function and a new calculation node rank method which can also be used for selecting the optimal path. Finally, evaluation results show that I-RPL outperforms ETXOF and several other improvements in terms of packet delivery ratio, latency, etc.

Diabetic nephropathy (DN) is a prevalent chronic microvascular complication associated with diabetes mellitus and represents a major cause of chronic kidney disease and renal failure. Current treatment strategies for DN primarily focus on symptom alleviation, lacking effective approaches to halt or reverse DN progression. Circular RNA (circRNA), characterized by a closed-loop structure, has emerged as a novel non-coding RNA regulator of gene expression, attributed to its conservation, stability, specificity, and multifunctionality. Dysregulation of circRNA expression is closely associated with DN progression, whereby circRNA impacts kidney cell injury by modulating cell cycle, differentiation, cell death, as well as influencing the release of inflammatory factors and stromal fibronectin expression. Consequently, circRNA is considered a predictive biomarker and a potential therapeutic target for DN. This review provides an overview of the latest research progress in the classification, functions, monitoring methods, and databases related to circRNA. The paper focuses on elucidating the impact and underlying mechanisms of circRNA on kidney cells under diabetic conditions, aiming to offer novel insights into the prevention, diagnosis, and treatment of DN.

The spreading of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) across the world has impacted people's health and lives worldwide in recent years. Rapid and accurate diagnosis is crucial for curbing the pandemic of coronavirus disease 2019 (COVID-19). Reverse transcription loop-mediated isothermal amplification (RT-LAMP) has great potential for SARS-CoV-2 detection but fails to completely replace conventional PCR due to the high false-positive rate (FPR). We proposed a triple-target RT-LAMP method for dual-signal, sensitive, and simultaneous detection of conserved genes of SARS-CoV-2. Multiple LAMP primer sets were designed for N, E, and M genes and their amplification efficacy were screened. Then, using artificial plasmids and RNA, the optimal primer set for each gene was examined on specificity, sensitivity, and detection range. The RT-LAMP initiated by these primer sets exhibited better specificity and sensitivity than that of RT-qPCR, and the tripletarget RT-LAMP could determine different variants of SARS-CoV-2. By testing 78 artificial RNA samples, the total FPR of triple-target RT-LAMP was eliminated compared with that of mono-target RT-LAMP. The tripletarget RT-LAMP method precisely identified throat swab specimens through colorimetry and fluorescent signals within 60 min, and the limit of detection (LOD) was as low as 187 copies/reaction. In the future, the tripletarget RT-LAMP can be applied to in-field and on-site diagnosis of symptomatic and asymptomatic virus carriers.

For the use in low-power and lossy networks (LLNs) under complex and harsh communication conditions, the routing protocol for LLNs (RPL) standardized by the Internet Engineering Task Force is specially designed. To improve the performance of LLNs, we propose a novel context-aware RPL algorithm based on a triangle module operator (CAR-TMO). A novel composite context-aware routing metric (CA-RM) is designed, which synchronously evaluates the residual energy index, buffer occupancy ratio of a node, expected transmission count (ETX), delay, and hop count from a candidate parent to the root. CA-RM considers the residual energy index and buffer occupancy ratio of the candidate parent and its preferred parent in a recursive manner to reduce the effect of upstream parents, since farther paths are considered. CA-RM comprehensively uses the sum, mean, and standard deviation values of ETX and delay of links in a path to ensure a better performance. Moreover, in CAR-TMO, the membership function of each routing metric is designed. Then, a comprehensive membership function is constructed based on a triangle module operator, the membership function of each routing metric, and a comprehensive context-aware objective function. A novel mechanism for calculating the node rank and the mechanisms for preferred parent selection are proposed. Finally, theoretical analysis and simulation results show that CAR-TMO outperforms several state-of-the-art RPL algorithms in terms of the packet delivery ratio and energy efficiency.

Glandular trichomes of tobacco (Nicotiana tabacum) produce blends of acylsucroses that contribute to defence against pathogens and herbivorous insects, but the mechanism of assembly of these acylsugars has not yet been determined. In this study, we isolated and characterized two trichome-specific acylsugar acyltransferases that are localized in the endoplasmic reticulum, NtASAT1 and NtASAT2. They sequentially catalyse two additive steps of acyl donors to sucrose to produce di-acylsucrose. Knocking out of NtASAT1 or NtASAT2 resulted in deficiency of acylsucrose; however, there was no effect on acylsugar accumulation in plants overexpressing NtASAT1 or NtASAT2. Genomic analysis and profiling revealed that NtASATs originated from the T subgenome, which is derived from the acylsugar-producing diploid ancestor N. tomentosiformis. Our identification of NtASAT1 and NtASAT2 as enzymes involved in acylsugar assembly in tobacco potentially provides a new approach and target genes for improving crop resistance against pathogens and insects.

Background: The regression of positive nodes in breast cancer after neoadjuvant chemotherapy (NAC) remains unknown. This study aimed to investigate this regression by injecting and tracing carbon nanoparticles (CNs) into the fusion node prior to NAC in patients with breast cancer. Methods: Guided by ultrasound, 0.3 mL of CNs suspension was injected in the fusion node prior to NAC in 110 patients with local advanced breast cancer. Then the patients underwent breast surgery and total axillary lymph node dissection following 2-6 cycles of NAC. The distribution by intercostobrachial nerves (ICBN) of positive nodes and black-stained nodes was researched, and the relationship between the distribution and lymphovascular invasion were investigated by response to NAC. Results: When patients were ranked by response to NAC (from sensitive to resistance), the number of positive nodes increased, as did the proportion of lymphovascular invasion, the number of black-stained nodes decreased. A significantly negative relationship was found between the number of positive nodes and the number of black-stained nodes (p < 0.001). The positive nodes in patients with sensitive consequence followed the rule from under the ICBN to above the ICBN. However, there was counterexample (skip metastasis) in the patients with resistance result. Conclusion: The regression of positive nodes follows the rule from upper to under, inner to outer in the patients with sensitive consequence to NAC. Long-term staining and tracing by CNs might provide an acceptable and feasible technique to investigate the regression of positive nodes, and would be a potential method for NAC-treated patients by using of ICBN. Trial registration: NCT 03355261. Retrospectively registered on November 28, 2017. (c) 2021 Asian Surgical Association and Taiwan Robotic Surgery Association. Publishing services by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/ licenses/by-nc-nd/4.0/).

Abstract Over 50% of all patients with cancer are treated with radiotherapy. However, radiotherapy is often insufficient as a monotherapy and requires a nontoxic radiosensitizer. Squalene epoxidase (SQLE) controls cholesterol biosynthesis by converting squalene to 2,3-oxidosqualene. Given that SQLE is frequently overexpressed in human cancer, this study investigated the importance of SQLE in breast cancer and non–small cell lung cancer (NSCLC), two cancers often treated with radiotherapy. SQLE-positive IHC staining was observed in 68% of breast cancer and 56% of NSCLC specimens versus 15% and 25% in normal breast and lung tissue, respectively. Importantly, SQLE expression was an independent predictor of poor prognosis, and pharmacologic inhibition of SQLE enhanced breast and lung cancer cell radiosensitivity. In addition, SQLE inhibition enhanced sensitivity to PARP inhibition. Inhibition of SQLE interrupted homologous recombination by suppressing ataxia-telangiectasia mutated (ATM) activity via the translational upregulation of wild-type p53-induced phosphatase (WIP1), regardless of the p53 status. SQLE inhibition and subsequent squalene accumulation promoted this upregulation by triggering the endoplasmic reticulum (ER) stress response. Collectively, these results identify a novel tumor-specific radiosensitizer by revealing unrecognized cross-talk between squalene metabolites, ER stress, and the DNA damage response. Although SQLE inhibitors have been used as antifungal agents in the clinic, they have not yet been used as antitumor agents. Repurposing existing SQLE-inhibiting drugs may provide new cancer treatments. Significance: Squalene epoxidase inhibitors are novel tumor-specific radiosensitizers that promote ER stress and suppress homologous recombination, providing a new potential therapeutic approach to enhance radiotherapy efficacy.

The objective of the study was to explore the role of SDC1 in breast cancer cells. Our study also investigated the regulatory relationship between SDC1 and the microRNA (miRNA) miR-335-5p as well as the impact of these two genes on the progression of breast cancer. Bioinformatic approaches were employed to analyze the differentially expressed messenger RNAs (mRNAs) and miRNAs (DE-mRNAs and DE-miRNAs) in breast cancer tissue. Then mRNA SC1 was obtained. Differentially downregulated mRNAs were intersected with target miRNAs predicted by databases, and miR-335-5p was determined as the study object. Quantitative reverse transcription polymerase chain reaction was applied to assess the expressions of SDC1 and miR-335-5p in each cell line. Next, Western blot assay was conducted to detect the protein level of SDC1 and dual-luciferase assay was performed to verify the binding relationship between miR-335-5p and SDC1. Finally, we conducted methyl thiazolyl tetrazolium (MTT), colony formation, and Transwell assays and flow cytometry to further investigate the impacts of SDC1 and miR-335-5p on the progression of breast cancer. SDC1 was significantly highly expressed while miR-335-5p was remarkably lowly expressed in human breast cancer. Silencing SDC1 in breast cancer blocked the proliferation, migration and invasion of the cells. In breast cancer, SDC1 was a target gene of miR-335-5p and silencing miR-335-5p notably increased SDC1 expression. Compared with the silence of miR-335-5p, simultaneous silences of miR-335-5p and SDC1 significantly reduced the proliferative, migratory and invasive abilities of breast cancer cells. The result revealed the interaction between miR-335-5p and SDC1 in the progression of breast cancer, which may contribute to the treatments for this cancer.

Purpose:It is well-known that the pathological complete response (pCR) rate in patients with luminal A cancer (LAC) is lower than those of other subtypes of breast cancer. The phenotype of cancer often alters after neoadjuvant chemotherapy (NAC) which may be related to hypoxia, and the latter might induce the drift of the estrogen receptor (ER). The phenotype drift in local advanced LAC after NAC might influence the long-term prognosis.Methods:The oxygen concentration of cancer tissues during NAC was recorded and analyzed (n = 43). The expression of ER and claudin-6 was detected in pre- and post-NAC specimens.Results:NAC might induce the cycling intracanceral hypoxia, and the pattern was related to NAC response. The median follow-up time was 61 months. Most of the patients (67%) with stable or increased ER and claudin-6 expression exhibited perfect prognosis (DFS = 100%, 61 months). About 20% of patients with decreased claudin-6 would undergo the poor prognosis (DFS = 22.2%, 61 months). The contrasting prognosis (100% vs. 22.2%) had nothing to do with the response of NAC in the above patients. Only 13% patients had stable claudin-6 and decreased ER, whose prognosis might relate to the response of NAC.Conclusion:NAC might induce cycling intracanceral hypoxia to promote the phenotype drift in local advanced LAC, and the changes in ER and claudin-6 after NAC would determine the long-term prognosis.

The integration of loop-mediated isothermal amplification (LAMP) onto a microfluidic chip adds more simplicity and portability for food safety control. However, this system for multiple-target detection was less used on foodborne bacteria. Here, we utilized a ten-well microfluidic chip with pre-loaded LAMP primer sets for the quantitative and qualitative detection of Salmonella, Staphylococcus aureus, Escherichia coli O157:H7, and Shigella. The specificity and sensitivity of designed primers targeting sirA, spA, fliC, and ipaH genes were validated. Using regular tubes and the ten-well microfluidic chips, these four pathogens could be simultaneously detected within 45 min by LAMP with fluorescent and colorimetric readouts, respectively. Finally, the results demonstrated that the limit of detection reached 8 x 10(3) CFU mL(-1) genomic DNA with a recovery rate of 86.1%similar to 110%. In summary, our microfluidic chip platform displayed obvious advantages in time-saving, cost-effectiveness, and sensitivity, which is applicable for in-field monitoring of food pathogens.

An intense terahertz laser field is shown to actively manipulate the electronic states, as well as the linear and nonlinear optical absorption coefficients, of the laterally-coupled quantum well wires (LCQWWs). The laser-dressed quantum states of the LCQWWs are achieved using the non-perturbative Floquet method and the two-dimensional diagonalization technique under the effective mass approximation. We have demonstrated that the intense terahertz laser field induces a strong deformation of the confinement potential configuration of the LCQWWs, thus pronouncedly dressing the energy levels and wave functions. An unambiguous picture is depicted for the evolution of the laser-dressed quantum states with the increase of the laser-dressed parameter characterizing the strength of the laser-dressed effect. On this basis, the resonant peak positions of the linear and nonlinear optical absorption coefficients feature a blue shift followed by a red shift with an increase of the laser-dressed parameter. Furthermore, the evolution of the peak values for the linear and third-order nonlinear optical absorption coefficients as a function of the laser-dressed parameter is comprehensively discussed. Moreover, in contrast to the case without intense terahertz laser field, the peak values of the linear, third-order nonlinear, and total optical absorption coefficients can be obviously enhanced at the same frequency position by manipulating the appropriate laser-dressed parameter. A similar feature can be found in the linear, third-order nonlinear, and total refractive index changes. Our findings are conducive to the implementation of the expected quantum states and nonlinear optical effects in the LCQWWs, paving the way for new designs in tunable optical switches, infrared photo-detectors and infrared modulators.

The inhibition of hydrogen peroxide (H2O2) on the cysteine-mediated aggregation of gold nanoparticles (AuNPs) has great potential to rapidly visualize H2O2 and disease-associated biomarkers. However, associated applications have been rigid due to the low cysteine-oxidation efficiency or the insufficient interference-resistance of previous catalysts. Here, we proposed a novel AuNPs colorimetric method termed Fe2+-catalyzed H2O2-preventing aggregation of AuNPs by oxidizing Cysteine (FeHOAuC) for the visible and rapid detection of H2O2 with high specificity. Formed Fe2+-cysteine composites in the beginning, Fe2+ accelerates the oxidation of cysteine by H2O2 with an intramolecular electron transferring model, and the generated cysteine oxides (including cysteic acid or cystine, dependent on the ratio of cysteine to H2O2 molecules) lose the pro-aggregation effect on AuNPs. Notably, FeHOAuC can quantify 2-30 mM of H2O2 within 5 min, and the monitoring process works well with tolerance to the impact of dissolved oxygen. The FeHOAuC paired with lactate oxidase was successfully used for measuring lactic acids in real sweat samples with a practicable detection window (2-60 mM) and a low variable coefficient (< 10%). In summary, FeHOAuC is a feasible platform for rapid and convenient detection of H2O2 and oxidase-specific substrates. (c) 2022 Elsevier B.V. All rights reserved.

With the national industrial upgrading and technological innovation in recent years, the construction industry is leading in the direction of informatization, industrialization, intelligence and international integration, which puts forward new requirements for the current traditional mode of computer talent training. The innovation of talent training mode, the improvement of education and teaching, the improvement of education resources and so on have become the urgent problems of new engineering computer professional talent training Absolutely. This paper analyzes the current situation of "new engineering" talent demand and training, points out the shortcomings of the current computer talent training in the teaching concept, teaching mode, teachers and so on, and explores the new engineering computer talent training mode. And take the practice of Henan University School of civil engineering and architecture in the new engineering personnel training as an example, hope to have a certain reference significance for the new engineering computer professional personnel training. Keywords: new engineering; computer; interdisciplinary training; subject integration DOI: 10.7176/JEP/12-8-03 Publication date: March 31 st 2021

针对缺乏科学合理的低功耗有损网络路由协议多路由度量评估方法,无法选择合适的下一跳,影响网络性能等问题,本文提出一种基于组合赋权法和逼近理想解排序法的多路由度量评估算法.该算法通过构建邻居节点各路由度量的初始判断矩阵,设计基于线性加权的复合目标函数,设计兼顾主客观因素的组合赋权算法确定复合目标函数中各路由度量的权重,并采用逼近理想解排序法确定下一跳节点等机制,有效地解决了上述问题.理论分析证明了该多路由度量评估算法的有效性和可靠性,仿真实验结果显示该算法在网络寿命,时延等方面均优于低功耗有损网络路由算法及其相关改进算法.

转化生长因子β (TGF-β)是促肝纤维的细胞因子,TGF-β潜态相关肽(LAP)和TGF-β的活化有关.本研究提取人血液中总RNA,逆转录转为cDNA,以此为模板PCR目的 片段,构建原核表达载体pET28a-LAP-B、pET28a-LAP-F,并在不同诱导温度、IPTG浓度、诱导时机下进行诱导表达,筛选最佳诱导条件,通过镍琼脂糖亲和层析纯化重组蛋白,获得的蛋白通过SDS-PAGE及Western blot鉴定,并检测重组蛋白的浓度.结果 表明,LAP全长及截短型原核表达载体构建成功,筛选最佳诱导条件为37℃培养4h,OD600值为0.8时降温至16℃,加入0.3 mmol/L (LAP-B)/0.45 mmol/L (LAP-F) IPTG诱导16h,大量培养后通过镍柱纯化和透析得到重组蛋白,经SDS-PAGE及Westem blot鉴定为LAP全长及截短型重组蛋白,并测得LAP-B、LAP-F重组蛋白浓度分别为637.6 μg/mL、126.6 μg/mL.本实验成功得到LAP全长及截短型重组蛋白,为研究LAP对TGF-β促纤维化作用的影响提供了实验基础.

BACKGROUND:Patients with early-stage luminal A breast cancer (LABC) have better prognoses. However, follow-up examinations are frequent and remain complex. The present study examined whether circulating tumor cell (CTC) detection could be used as an earlier and more reproducible indicator of disease status among patients with early-stage LABC, and given China's healthcare resource challenges, whether it could periodically replace follow-up routine imaging.METHODS:A total of 135 postoperative Chinese patients with early-stage LABC were randomly assigned to a CTC group (68 patients underwent alternating assessments using CTC detection and routine re-examination) or control group (67 patients underwent only routine re-examination). The prognosis and patient-covered costs of the various assessments were calculated for the 2 groups.RESULTS:No patients had normal CTCs and simultaneous abnormal imaging findings. There were no differences in overall survival, disease-free survival and total patient-covered cost of follow-up between the 2 groups (all P > 0.05). However, there was a significant difference in the average patient-covered cost (P < 0.001). Furthermore, significant intergroup differences were observed in the total and average hospitalization times (P < 0.05).CONCLUSIONS:Among Chinese patients with low-risk LABC, CTC detection was highly reliable and relatively low cost. Therefore, CTC detection may be used to reduce the number of routine imaging follow-ups.

After the development of deep learning object tracking methods in recent years, the fully convolutional siamese network object tracking algorithm SiamFC has become a more classic deep learning object tracking algorithm. In view of the problem that the accuracy of the tracking results of SiamFC will be reduced in the case of complex backgrounds, this paper introduces the attention mechanism based on the SiamFC, which performs channel and spatial weighting on the feature maps obtained by convolution of the input image. At the same time, the backbone network model of CNN in the algorithm is adjusted, then the siamese network combined with attention mechanism for object tracking is proposed. It can strengthen the effectiveness of the results of feature extraction and enhance the ability of the network model to discriminate targets. In this paper, the algorithm is tested on the OTB2015, VOT2016 and VOT2017 datasets, and compared with multiple object tracking algorithms. Experimental results show that the algorithm in this paper can better solve the complex background problem in object tracking, and has certain advantages compared with other algorithms.