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Long noncoding RNAs (lncRNAs) and promoter- or enhancer-associated unstable transcripts locate preferentially to chromatin, where some regulate chromatin structure, transcription and RNA processing1-13. Although several RNA sequences responsible for nuclear localization have been identified-such as repeats in the lncRNA Xist and Alu-like elements in long RNAs14-16-how lncRNAs as a class are enriched at chromatin remains unknown. Here we describe a random, mutagenesis-coupled, high-throughput method that we name 'RNA elements for subcellular localization by sequencing' (mutREL-seq). Using this method, we discovered an RNA motif that recognizes the U1 small nuclear ribonucleoprotein (snRNP) and is essential for the localization of reporter RNAs to chromatin. Across the genome, chromatin-bound lncRNAs are enriched with 5' splice sites and depleted of 3' splice sites, and exhibit high levels of U1 snRNA binding compared with cytoplasm-localized messenger RNAs. Acute depletion of U1 snRNA or of the U1 snRNP protein component SNRNP70 markedly reduces the chromatin association of hundreds of lncRNAs and unstable transcripts, without altering the overall transcription rate in cells. In addition, rapid degradation of SNRNP70 reduces the localization of both nascent and polyadenylated lncRNA transcripts to chromatin, and disrupts the nuclear and genome-wide localization of the lncRNA Malat1. Moreover, U1 snRNP interacts with transcriptionally engaged RNA polymerase II. These results show that U1 snRNP acts widely to tether and mobilize lncRNAs to chromatin in a transcription-dependent manner. Our findings have uncovered a previously unknown role of U1 snRNP beyond the processing of precursor mRNA, and provide molecular insight into how lncRNAs are recruited to regulatory sites to carry out chromatin-associated functions.

OBJECTIVES: To compare the dynamic visual acuity (DVA) following implantation of trifocal with monofocal intraocular lenses (IOL) and using a novel test system. METHODS: The present research was a retrospective, multicenter clinical study. Two hundred and ten eyes of 149 patients that underwent cataract phacoemulsification and IOL implantation were enrolled. One hundred and ten eyes of patients received trifocal (AT LISA tri839MP, Carl Zeiss Meditec, Germany) and 100 eyes received monofocal (Tecnis ZCB00, Abbott, United States) lenses and were evaluated 3 months after implantation. Outcome measures included monocular uncorrected distance (UDVA), intermediate (UIVA) and near (UNVA) visual acuity and best corrected distance visual acuity (BCDVA; logMAR units); contrast sensitivity under photopic, mesopic, with glare conditions; and dynamic visual acuity using a self-developed system. RESULTS: There was no statistically significant difference in baseline characteristics between groups. Monocular UDVA, UIVA, and UNVA were significantly better (all p

A novel metal preloading route is proposed to prepare Pt/SAPO-11 bifunctional catalysts. The addition of platinum brings about an increase in catalyst product yield with a growth rate of 87% attributed to chemical adsorption between Pt and the lone pair of electrons in the nitrogen atom of the structure-directing agent. More interestingly, it can be concluded from the HAADF-STEM, SE-STEM images and DRIFTS data on CO adsorption, there are small Pt nanoparticles confined inside the SAPO-11 molecular sieve, while the big Pt nanoparticles are dispersed on the surface. The synergistic effect between the metal sites and acid sites can be improved by the enhancement of metal–acid balance, by adjusting the nPt/nA ratio. A better metal–acid balance property is achieved and the carbenium ion can quickly go through the hydrogenating reaction on Pt nanoparticles, hence shortening the time it is trapped in the acid sites upon n-dodecane hydroisomerization. Consequently, the cracking reaction can be effectively avoided and liquid yield as high as 99% and isomer yield up to 84% is produced, much superior than the Pt/SAPO-11 catalyst synthesized via the traditional impregnation method of Pt with the maximum isomer yield of 52%. Moreover, more multi-branched isomers are produced and can further help improve the low-temperature rheology when used as lubricant base oil.

RNA polymerase II (Pol II)-mediated transcription in eukaryotic cells starts with assembly of preinitiation complex (PIC) on core promoter, a DNA sequence of ∼100 base pairs. The transcription PIC consists of Pol II and general transcription factors TFIID, TFIIA, TFIIB, TFIIF, TFIIE, and TFIIH. Previous structural studies focused on PIC assembled on TATA box promoters with TFIID replaced by its subunit, TATA box-binding protein (TBP). However, the megadalton TFIID complex is essential for promoter recognition, TBP loading onto promoter, and PIC assembly for almost all Pol II-mediated transcription, especially on the TATA-less promoters, which account for ∼85% of core promoters of human coding genes. The functions of TFIID could not be replaced by TBP. The recent breakthrough in structure determination of TFIID-based PIC complexes in different assembly stages revealed mechanistic insights into PIC assembly on TATA box and TATA-less promotes and provided a framework for further investigation of transcription initiation.

DNA methyltransferases (DNMTs) catalyze DNA methylation, and their functions in mammalian embryonic development and diseases including cancer have been extensively studied. However, regulation of DNMTs remains under study. Here, we show that CCAAT/enhancer binding protein α (CEBPA) interacts with the long splice isoform DNMT3A, but not the short isoform DNMT3A2. CEBPA, by interacting with DNMT3A N-terminus, blocks DNMT3A from accessing DNA substrate and thereby inhibits its activity. Recurrent tumor-associated CEBPA mutations, such as preleukemic CEBPA N321D mutation, which is particularly potent in causing AML with high mortality, disrupt DNMT3A association and cause aberrant DNA methylation, notably hypermethylation of PRC2 target genes. Consequently, leukemia cells with the CEBPA N321D mutation are hypersensitive to hypomethylation agents. Our results provide insights into the functional difference between DNMT3A isoforms and the regulation of de novo DNA methylation at specific loci in the genome. Our study also suggests a therapeutic strategy for the treatment of CEBPA -mutated leukemia with DNA-hypomethylating agents.

The study aimed to investigate the role of androgen receptor (AR)/cell cycle-related kinase (CCRK) signalling pathway in chronic hepatitis B virus (HBV) infection and gender differences, and the contribution of AR regulatory factor signal transducer and activator of transcription 3 (STAT3) in it. AR, CCRK, and phosphorylated STAT3 expressions in liver tissues of chronic HBV-infected patients and non-HBV controls were determined by western blot and compared between genders. The relationships of expression levels with serum HBV DNA levels, liver inflammation activity, and fibrosis score were analysed in chronic HBV-infected patients. The relationships between expression levels of three proteins were also analysed. HBV-infected patients had significantly higher expression levels of AR, CCRK, and p-STAT3

The CRISPR/Cas9 system is a revolutionary genome editing technique and has been widely used in numerous plants. For plants (e.g. citrus) with very low transformation efficiency, how to optimize gene editing efficiency and induce large-fragment deletion has been the focus of research. Here, we report that CRISPR/Cas9 induces efficient deletion of 16-673 bp fragments in the genome of

Purpose: Camrelizumab, an mAb against programmed cell death protein 1 (PD-1), plus nab-paclitaxel exhibited promising antitumor activity in refractory metastatic immunomodulatory triple-negative breast cancer (TNBC). Famitinib is a tyrosine kinase inhibitor targeting VEGFR2, PDGFR, and c-kit. We aimed to assess the efficacy and safety of a novel combination of famitinib, camrelizumab, and nab-paclitaxel in advanced immunomodulatory TNBC. Patients and Methods: This open-label, single-arm, phase II study enrolled patients with previously untreated, advanced, immunomodulatory TNBC (CD8 IHC staining ≥10%). Eligible patients received 20 mg of oral famitinib on days 1 to 28, 200 mg of i.v. camrelizumab on days 1 and 15, and i.v. nab-paclitaxel 100 mg/m2 on days 1, 8, and 15 in 4-week cycles. The primary endpoint was objective response rate (ORR), as assessed by investigators per RECIST v1.1. Key secondary endpoints were progression-free survival (PFS), overall survival (OS), duration of response (DOR), safety, and exploratory biomarkers. Results: Forty-eight patients were enrolled and treated. Median follow-up was 17.0 months (range, 8.7–24.3). Confirmed ORR was 81.3% [95% confidence interval (CI), 70.2–92.3], with five complete and 34 partial responses. Median PFS was 13.6 months (95% CI, 8.4–18.8), and median DOR was 14.9 months [95% CI, not estimable (NE)–NE]. Median OS was not reached. No treatment-related deaths were reported. Among 30 patients with IHC, 13 (43.3%) were programmed death-ligand 1 (PD-L1)–negative, and PD-L1 was associated with favorable response. PKD1 and KAT6A somatic mutations were associated with therapy response. Conclusions: The triplet regimen was efficacious and well tolerated in previously untreated, advanced, immunomodulatory TNBC. The randomized controlled FUTURE-SUPER trial is under way to validate our findings. See related commentary by Salgado and Loi, p. 2728

The higher extraction efficiency of analytes is crucial for developing immunoassays with high accuracy. Here, we evaluated the extraction efficiency of neonicotinoids in tea samples in terms of grinding degrees, extraction solvents types and contents. Fragments for fresh tea leaves (1 g, 5-10 mm

Xiaodan Jiang,1,2,* Yu Wan,1,2,* Hao Yuan,1,2,* Liming Zhao,3,* Min Sun,4,* Yanhui Xu,5,* Xiangyang Xin,6,* Jing Dong,7,* Die Hu,8,* Dongmei Chen,9,* Xuemin Li1,2 1Department of Ophthalmology, Peking University Third Hospital, Beijing, People’s Republic of China; 2Beijing Key Laboratory of Restoration of Damaged Ocular Nerve, Peking University Third Hospital, Beijing, People’s Republic of China; 3Department of Ophthalmology, Beijing Fengtai Hospital, Beijing, People’s Republic of China; 4Department of Ophthalmology, Huabei Petroleum General Hospital, Renqiu, Hebei, People’s Republic of China; 5Department of Ophthalmology; Hebei Provincial Eye Hospital, Xingtai, Hebei, People’s Republic of China; 6Department of Ophthalmology, Inner Mongolia Baogang Hospital, Baotou, Inner Mongolia, People’s Republic of China; 7Department of Ophthalmology, the First Affiliated Hospital of Baotou Medical College, Baotou, Inner Mongolia, People’s Republic of China; 8Department of Ophthalmology, Baoding Zhuozhou GEM Flower Hospital, Zhuozhou, Hebei, People’s Republic of China; 9Department of Ophthalmology, Baoding GEM Flower Eastern Hospital, Zhuozhou, Hebei, People’s Republic of China*These authors contributed equally to this workCorrespondence: Xuemin Li, Department of Ophthalmology, Beijing Key Laboratory of Restoration of Damaged Ocular Nerve, Peking University Third Hospital, No. 49 Huayuan North Road, Haidian District, Beijing, 100191, People’s Republic of China, Tel +86 10 8226 6312, Fax +86 10 8208 9951, Email lxmlxm66@sina.comObjective: To investigate the incidence of acute postoperative endophthalmitis (POE) after cataract surgery in Northern China from 2013 to 2019, evaluate the efficacy of prophylaxis and analyze the predictors of visual prognosis among POE patients.Methods: The study was conducted as a retrospective multi-center research, with seven hospitals in Northern China enrolled. The diagnosis of acute-onset POE was made on the basis of clinical manifestations within six weeks after initial surgery. By reviewing electronic medical system, the number of cataract surgeries and acute POE cases were recorded to estimate the overall incidence and incidences by different years and hospitals. Perioperative measures for preventing infection in different hospitals were collected. The correlations between unfavorable final vision and potential factors including basic information and clinical characteristics were examined to determine the predictive factors for final visual prognosis.Results: Of 72,255 cataract surgeries performed during seven years in the seven hospitals, 19 cases developed acute POE, yielding an overall incidence of 0.026%. The average incidence of acute POE among seven hospitals significantly declined annually during the past 7 years (p = 0.021). In Hospital-D, the incidence of acute POE significantly decreased after the application of 0.5% povidone-iodine (PVP-I) for conjunctival washing (p = 0.003). Two hospitals adopting tobramycin in the irrigation solution achieved a significant lower incidence of POE than the other hospitals (p = 0.044). The positive rate of pathogen culture was just 17.6% (3/19). Patients with presenting BCVA of CF or better were more likely to present with unfavorable final vision than those with worse presenting BCVA (p = 0.003).Conclusion: The overall incidence of acute POE after cataract surgery from 2013 to 2019 in Northern China was 0.026%, and the incidence declined annually over the period. Presenting BCVA could be a significant prognosis factor for predicting the final visual outcomes of acute POE patients.Keywords: endophthalmitis, cataract surgery, infection prophylaxis, prognosis predictors

In this paper, a secure and low energy dynamic slicing algorithm, namely, Improved D-SMART (IM-D-SMART) based on the Data Aggregation Protocol on Slice Mix Aggregate (D-SMART) is proposed to improve the security and confidentiality of wireless sensor networks and reduce the energy consumption of nodes in data collection and transmission in wireless sensor networks. According to the importance of data, the residual energy of nodes and the relative density of nodes, the data are dynamically partitioned to improve the D-SMART algorithm. Simultaneously, sending negative number splicing is used to compensate for the loss caused by the collision of data transmission between nodes. The simulation results show that IM-D-SMART outperforms D-SMART in terms of computation, privacy and communication cost.

Background Biliary atresia (BA) is a severe cholangiopathy possibly resulting from virus-induced and immune-mediated injury of the biliary system. IFN-γ, secreted from CD4+ Th1 cells and CD8+ cytotoxic T cells, is a major mediator of liver pathology. Programmed death protein-1 (PD-1) signaling suppresses T cell function. However, how PD-1 modify T cell function in BA remains incompletely understood. Methods Frequencies of PD-1 expressing CD4+ and CD8+ T cells were analyzed in the liver and blood from BA and control subjects. Associations of PD-1+CD4+/CD8+T cell abundances with liver function indices were measured. Function of PD-1 was measured by administration of an anti-PD-1 antibody in a Rhesus Rotavirus (RRV)-induced BA model. Survival, histology, direct bilirubin, liver immune cell subsets and cytokine production were analyzed. Results PD-1 was significantly upregulated in CD4+ and CD8+ T cells in patients with BA compared with control subjects. PD-1 expression in T cells was negatively associated with IFN-γ concentration in liver (PD-1+CD4+T cells in liver vs. IFN-γ concentration, r = − 0.25, p = 0.05; PD-1+CD8+T cells in liver vs. IFN-γ concentration, r = − 0.39, p = 0.004). Blockade of PD-1 increased IFN-γ expression in CD4+ T and CD8+ T cells (RRV vs. anti-PD-1 treated RRV mice: 11.59 ± 3.43% vs. 21.26 ± 5.32% IFN-γ+ in hepatic CD4+T cells, p = 0.0003; 9.33 ± 4.03% vs. 22.55 ± 7.47% IFN-γ+ in hepatic CD8+T cells, p = 0.0001), suppressed bilirubin production (RRV vs. anti-PD-1 treated RRV mice: 285.4 ± 47.93 vs. 229.8 ± 45.86 μmol/L total bilirubin, p = 0.01) and exacerbated liver immunopathology. Conclusions PD-1 plays a protective role in infants with BA by suppressing IFN-γ production in T cells. Increasing PD-1 signaling may serve as a therapeutic strategy for BA.

5-Methylcytosine is found in both DNA and RNA; although its functions in DNA are well established, the exact role of 5-methylcytidine (m(5)C) in RNA remains poorly defined. Here we identified, by employing a quantitative proteomics method, multiple candidate recognition proteins of m(5)C in RNA, including several YTH domain-containing family (YTHDF) proteins. We showed that YTHDF2 could bind directly to m(5)C in RNA, albeit at a lower affinity than that toward N(6)-methyladenosine (m(6)A) in RNA, and this binding involves Trp(432), a conserved residue located in the hydrophobic pocket of YTHDF2 that is also required for m(6)A recognition. RNA bisulfite sequencing results revealed that, after CRISPR-Cas9-mediated knockout of the YTHDF2 gene, the majority of m(5)C sites in rRNA (rRNA) exhibited substantially augmented levels of methylation. Moreover, we found that YTHDF2 is involved in pre-rRNA processing in cells. Together, our data expanded the functions of the YTHDF2 protein in post-transcriptional regulations of RNA and provided novel insights into the functions of m(5)C in RNA biology.

Glycolytic enzyme phosphoglycerate mutase 1 (PGAM1) plays a critical role in cancer metabolism by coordinating glycolysis and biosynthesis. A well-validated PGAM1 inhibitor, however, has not been reported for treating pancreatic ductal adenocarcinoma (PDAC), which is one of the deadliest malignancies worldwide. By uncovering the elevated PGAM1 expressions were statistically related to worse prognosis of PDAC in a cohort of 50 patients, we developed a series of allosteric PGAM1 inhibitors by structure-guided optimization. The compound KH3 significantly suppressed proliferation of various PDAC cells by down-regulating the levels of glycolysis and mitochondrial respiration in correlation with PGAM1 expression. Similar to PGAM1 depletion, KH3 dramatically hampered the canonic pathways highly involved in cancer metabolism and development. Additionally, we observed the shared expression profiles of several signature pathways at 12 h after treatment in multiple PDAC primary cells of which the matched patient-derived xenograft (PDX) models responded similarly to KH3 in the 2 wk treatment. The better responses to KH3 in PDXs were associated with higher expression of PGAM1 and longer/stronger suppressions of cancer metabolic pathways. Taken together, our findings demonstrate a strategy of targeting cancer metabolism by PGAM1 inhibition in PDAC. Also, this work provided “proof of concept” for the potential application of metabolic treatment in clinical practice.

In this communication, a differentially fed wideband slot antenna on a single layer with endfire radiation and filtering response under multi-resonant modes is proposed. By using the $\lambda $ /4 slot radiator with its opened terminal at one side, the endfire-radiated fields of the slot antenna is initially formed with compact dimension and endfire radiation. After that, the $\lambda $ /4 slot radiator with two distinctive portions of different slot widths is presented and investigated. The results demonstrate that the resonant frequencies of the first two odd-order modes could be progressively reallocated in proximity to each other as the length and width ratios of these two portions are properly selected. Thus, the bandwidth in input impedance of the entire differentially fed slot antenna could be significantly improved under operation of dual-resonant modes. Next, an extra microstrip-line resonator is introduced in the feeding part so as to produce a third resonant mode and improve its frequency selectivity in the wide operating band. To validate these predicted performances, a filtering slot antenna operating from 3.33 to 4.50 GHz is in final fabricated and tested. The measured results exhibit good impedance matching property, flat radiation gain of about 5 dBi, and improved endfire radiation characteristic with the front-to-back ratio (FBR) better than 9.0 dB in the operating band. Both the simulated and measured results are provided with good agreement.

Doxorubicin (Dox) is a widely used fluorescent chemotherapy drug. Its primary delivery systems, based on physical adsorption to silica nanoparticles, can lead to low drug loading. Direct loading of Dox via covalent bonds during the formation of silica nanoparticles has never been reported. In this work, we designed and synthesized a silane-borate precursor, which contains not only an alkoxysilane moiety to form organosilica nanoparticles but also a phenylboronic acid moiety to react with diol-containing compounds. Using this compound, the covalent loading of Dox during the preparation of organosilica nanoparticles was effectively realized with a high drug loading content up to 22.4 %. Further modification by hyaluronic acid (HA) bestowed the Si-Dox@HA nanoparticles with the ability to target CD44-overexpressing cancer cells. The Si-Dox@HA nanoparticles exhibited H2 O2 -responsive release of about 80 % Dox and displayed seven-fold selectivity for killing cancer cells over normal cells, relative to Dox and Si-Dox nanoparticles. Moreover, these Si-Dox@HA nanoparticles are also suitable for targeted fluorescence imaging of CD44-overexpressing cancer cells.

Alkaline stress has serious-negative effects on citrus production. Ziyang xiangcheng (Citrus junos Sieb. ex Tanaka) (Cj) is a rootstock that is tolerant to alkaline stress and iron deficiency. Trifoliate orange (Poncirus trifoliata (L.) Raf.) (Pt), the most widely used rootstock in China, is sensitive to alkaline stress. To investigate the molecular mechanism underlying the tolerance of Cj to alkaline stress, next-generation sequencing was employed to profile the root transcriptomes and small RNAs of Cj and Pt seedlings that were cultured in nutrient solutions along a three pH gradient. This two-level regulation data set provides a system-level view of molecular events with a precise resolution. The data suggest that the auxin pathway may play a central role in the inhibitory effect of alkaline stress on root growth and that the regulation of auxin homeostasis under alkaline stress is important for the adaptation of citrus to alkaline stress. Moreover, the jasmonate (JA) pathway exhibits the opposite response to alkaline stress in Cj and Pt and may contribute to the differences in the alkaline stress tolerance and iron acquisition between Cj and Pt. The dataset provides a wealth of genomic resources and new clues to further study the mechanisms underlying alkaline stress resistance in Cj., Molecular biology: Rooting out alkaline resistance mechanisms in citrus trees The ability of citrus trees to tolerate alkaline soils may hinge upon plant hormones called auxins which regulate root growth. The discovery could aid the development of more resilient rootstocks. Alkaline stress, caused by e.g. industrial run-off, is a growing problem worldwide because it reduces the growth and survival of crops – including the most widely used citrus rootstock in China, Poncirus trifoliata. To better understand the mechanisms underpinning tolerance to soil alkalinity, Hualin Yi at Huazhong Agricultural University in Wuhan, China, and colleagues used next-generation sequencing to profile the transcription products of P. trifolata seedlings and those of an alkaline-tolerant rootstock, Ziyang xiangcheng, when they were grown in three different nutrient solutions. Auxin homeostasis appears to be a key element of citrus adaption to alkaline stress, they found - probably by encouraging lateral root branching.

A facile strategy was proposed to improve electrochemical performances of LiEuTiO4 by surface modification via pyrolysis of urea at a rather low temperature of 400 °C in N2 atmosphere. The modified LiEuTiO4 exhibited excellent electrochemical performances, including high capacity (166.3 mA h g−1 at 100 mA g−1 at 1st cycle), high rate capability (129.4 mA h g−1 at 1600 mA g−1), and long cyclic stability (149.8 mA h g−1 after 650 cycles at 500 mA g−1). The effects of surface modification on structure and electrochemical performances were extensively studied. SEM and TEM results indicated the pyrolysis of urea surface modification which successfully lead to the formation of a nitrogen–carbon co-existed coating layer. XPS analysis confirmed the presence of N-doped carbon and TiN, which were attributed to remarkable reduction of the polarization and enhancement of the conductivity of LiEuTiO4.

This study investigated the effects of ecological-level marketization, individual-level occupational status, and their interaction, on depression in residents in urban China. Population-based data ( N = 13,004) from the 2016 China Family Panel Survey were used. A multilevel mixed-effects generalized linear model explored whether and to what extent market transition measured by the marketization index (MI), occupational status measured by international socio-economic index (ISEI), and their interaction, affected people’s depression. Results showed that higher MI ( b = –.157, p < .001) and ISEI scores ( b = –.124, p < .001) were associated with lower levels of depression. However, residents with high occupational status might suffer a uniquely elevated level of depression when living in highly marketized cities ( b = .139, p < .05). Raising the public mental health awareness of residents with low occupational status from low marketized areas and that of residents with high occupational status from high marketized areas is warranted in societies undergoing rapid marketization, such as China.

Emerging building analytics rely on data-driven machine learning algorithms. However, writing these analytics is still challenging---developers not only need to know what data are required by the analytics but also how to reach the data in each individual building, despite the existing solutions to standardizing data and resource management in buildings. To bridge the gap between analytics development and the specific details of reaching the actual data in each building, we present Energon, an open-source system that enables portable building analytics. The core of Energon is a new data organization of building data, as well as the tools that can effectively manage building data and support building analytics development. More specifically, we propose a new "logic partition" of data resources in buildings, and this abstraction universally applies to all buildings. We develop a declarative query language to find data resources in this new logic views with high-level queries, thus substantially reducing development efforts. We also develop a query engine with automatic data extraction by traversing building ontology that widely exists in buildings. In this way, Energon enables analytics requirements to be mapped to building resources in a building-agnostic manner. Using four types of real-world building analytics, we demonstrate the use of Energon as well as its effectiveness in reducing development efforts.

RNA polymerase II (Pol II)-mediated transcription in metazoan requires precise regulation. RNA polymerase II-associated protein 2 (RPAP2) was previously identified to transport Pol II from cytoplasm to nucleus and dephosphorylates Pol II C-terminal domain (CTD). We found that RPAP2 binds hypo/hyper-phosphorylated Pol II with undetectable phosphatase activity. Structure of RPAP2-Pol II shows mutually exclusive assembly of RPAP2-Pol II and pre-initiation complex (PIC) due to three steric clashes. RPAP2 prevents/disrupts Pol II-TFIIF interaction and impairs in vitro transcription initiation, suggesting a function in prohibiting PIC assembly. Loss of RPAP2 in cells leads to global accumulation of TFIIF and Pol II at promoters, indicating critical role of RPAP2 in inhibiting PIC assembly independent of its putative phosphatase activity. Our study indicates that RPAP2 functions as a gatekeeper to prohibit PIC assembly and transcription initiation and suggests a novel transcription checkpoint.

YAP and TAZ (YAP/TAZ), two major effectors of the Hippo signaling pathway, are frequently activated in human cancers. The activity of YAP/TAZ is strictly repressed upon phosphorylation by LATS1/2 tumor suppressors. However, it is unclear how LATS1/2 are precisely regulated by upstream factors such as Hippo kinases MST1/2. Here, we show that WWC proteins (WWC1/2/3) directly interact with LATS1/2 and SAV1, and SAV1, in turn, brings in MST1/2 to phosphorylate and activate LATS1/2. Hence, WWC1/2/3 play an organizer role in a signaling module that mediates LATS1/2 activation by MST1/2. Moreover, we have defined a minimum protein interaction interface on WWC1/2/3 that is sufficient to activate LATS1/2 in a robust and specific manner. The corresponding minigene, dubbed as SuperHippo, can effectively suppress tumorigenesis in multiple tumor models. Our study has uncovered a molecular mechanism underlying LATS1/2 regulation and provides a strategy for treating diverse malignancies related to Hippo pathway dysregulation.

This letter presents a novel multilayer dual-band balanced bandpass filter (BPF) design by using two perturbed circular patch resonators. The TM11 mode and TM21 mode of the resonator with odd-symmetric field distributions are explored to realize the desired differential-mode (DM) transmission and common-mode (CM) suppression. Two circular patches are properly coupled in the back-to-back form to realize a dual-passband balanced response by virtue of coupling apertures etched on the ground. In addition to the internal coupling, the above apertures are also further utilized for the undesired degenerate mode harmonic suppression. Besides, slot perturbations on the patch are introduced to perturb the TM21 resonant mode to independently adjust the center frequency of the higher passband, while the lower passband remains almost unchanged. Thus, two passbands can be flexibly controlled by simultaneously tuning the above slots and size of the patch. For validation, a dual-band balanced BPF prototype is implemented. The results indicate 18 and 26% wide fractional bandwidths centered at 5.5 and 7.5 GHz with return loss higher than 20 dB under DM operation and CM suppression higher than 40 dB over an ultra-wide frequency band.

Background: Biliary atresia (BA) is a severe cholangiopathy resulting from virus-induced and immune-mediated injury of the biliary system. IFN-g, secreted from CD4+ Th1 cells and CD8+ cytotoxic T cells, is a major mediator of liver pathology. Programmed death 1 (PD-1) signaling suppresses T cell function. However, how PD-1 modify T cell function in BA remains incompletely understood. Methods: Frequencies of PD-1 expressing CD4+ and CD8+ T cells were analyzed in the liver and blood from BA and control subjects. Associations of PD-1+CD4+/CD8+T cell abundances with liver function indices were measured. Function of PD-1 was measured by administration of an anti-PD-1 antibody in a Rhesus Rotavirus (RRV)-induced BA model. Survival, histology, direct bilirubin, liver immune cell subsets and cytokine production were analyzed. Results: PD-1 was significantly upregulated in CD4+ and CD8+ T cells in patients with BA compared with control subjects. PD-1 expression in T cells was negatively associated with IFN-γ concentration in liver. Blockade of PD-1 increased IFN-g expression in CD4+ T and CD8+ T cells, suppressed bilirubin production and exacerbated liver immunopathology.Conclusions: PD-1 plays a protective role in infants with BA by suppressing IFN-g production in T cells. Increasing PD-1 signaling may serve as a therapeutic strategy for BA.

Non-small cell lung cancer (NSCLC) has high morbidity and mortality rates worldwide, and tumor metastasis is generally associated with poor prognosis. Chemotherapy resistance aggravates the challenges associated with treating NSCLC. Therefore, identifying effective targets and developing therapies based on these findings could bring novel perspectives for patients with metastatic NSCLC. The expression levels of receptor tyrosine-protein kinase erbB-2 (ERBB2) are associated with NSCLC progression. Differential microRNA (miR) expression profiles have been identified in tumors and can be used to identify multiple malignant phenotypes. miR-133a-3p expression is dysregulated in a variety of tumors. However, to the best of our knowledge, the association between miR-133a-3p and the NSCLC pathogenesis process has not been demonstrated yet. The present study revealed a decrease in miR-133a-3p expression in both tissues and cell lines, which was detected using reverse transcription-quantitative (RT-q)PCR, and western blotting and RT-qPCR demonstrated ERBB2 levels were increased at both protein and mRNA levels. Bioinformatics analysis and dual-luciferase reporter assays demonstrated that ERBB2 was a direct target of miR-133a-3p. Furthermore, MTT, wound healing and Transwell assays revealed that overexpression of miR-133a-3p suppressed proliferation, invasion and migration of NSCLC cells, respectively, effects that were inhibited following ERBB2 overexpression. In addition, immunofluorescence assays demonstrated that overexpression of ERBB2 upregulated N-cadherin expression, while E-cadherin expression was downregulated. In conclusion, the present data demonstrated that miR-133a-3p acted as a tumor suppressor by negatively regulating ERBB2 expression. The miR-133a-3p/ERBB2 axis may be a potential target for the diagnosis and treatment of NSCLC in the future.

An RNA-involved phase-separation model has been proposed for transcription control. However, the molecular links that connect RNA to the transcription machinery remain missing. Here we find that RNA-binding proteins (RBPs) constitute half of the chromatin proteome in embryonic stem cells (ESCs), some being colocalized with RNA polymerase (Pol) II at promoters and enhancers. Biochemical analyses of representative RBPs show that the paraspeckle protein PSPC1 inhibits the RNA-induced premature release of Pol II, and makes use of RNA as multivalent molecules to enhance the formation of transcription condensates and subsequent phosphorylation and release of Pol II. This synergistic interplay enhances polymerase engagement and activity via the RNA-binding and phase-separation activities of PSPC1. In ESCs, auxin-induced acute degradation of PSPC1 leads to genome-wide defects in Pol II binding and nascent transcription. We propose that promoter-associated RNAs and their binding proteins synergize the phase separation of polymerase condensates to promote active transcription.

RNA polymerase II (Pol II)-mediated transcription in metazoans requires precise regulation. RNA Pol II-associated protein 2 (RPAP2) was previously identified to transport Pol II from cytoplasm to nucleus and dephosphorylates Pol II C-terminal domain (CTD). Here, we show that RPAP2 binds hypo-/hyper-phosphorylated Pol II with undetectable phosphatase activity. The structure of RPAP2-Pol II shows mutually exclusive assembly of RPAP2-Pol II and pre-initiation complex (PIC) due to three steric clashes. RPAP2 prevents and disrupts Pol II-TFIIF interaction and impairs in vitro transcription initiation, suggesting a function in inhibiting PIC assembly. Loss of RPAP2 in cells leads to global accumulation of TFIIF and Pol II at promoters, indicating a critical role of RPAP2 in inhibiting PIC assembly independent of its putative phosphatase activity. Our study indicates that RPAP2 functions as a gatekeeper to inhibit PIC assembly and transcription initiation and suggests a transcription checkpoint.

Purpose. To evaluate the short-term safety and efficacy of a novel approach of utilizing the 9-0 looped polypropylene suture with double knots buried into the scleral groove and the scleral tunnel to minimize the risk of the suture erosion and suture knot exposure. Design. Clinical-based retrospective study. Methods. Records of consecutive patients who had anterior vitrectomy and scleral-fixated posterior chamber intraocular lens (IOL) implantation between July 2018 and April 2020 with a minimum follow-up of 3 months were reviewed. Results. This study enrolled a total of 21 eyes from 20 patients (15 male). These patients had a mean age of 58.52 ± 8.55 years and were followed for an average of 1.08 ± 0.58 years postoperatively. Best-corrected visual acuity (BCVA) improved from a preoperative mean of 0.43 ± 0.41 logMAR to a significantly higher mean 3-month postoperative value of 0.09 ± 0.21 logMAR (Z = -3.35, p < 0.01 ). There were no statistical differences between the preoperative and postoperative corneal endothelial cell density ( p = 0.71 ). The postoperative complications included transient increased intraocular pressure in 5 eyes (24%). No other complications were detected during the follow-up. Conclusions. The modified technique proposed is a safe, effective, and reliable approach resulting in good visual outcomes. Our procedure might have the potential benefit to avoid suture-related complications in scleral-fixated IOL implantation. Trial registration. Retrospective case series study, not applicable.

This paper has proposed an end-fire slot antenna with back-lobe level suppression and front-to-back ratio (FBR) improvement. At first, the radiation characteristic of the conventional λ/4 slot antenna are meticulously investigated to initially demonstrate that its FBR can be maintained as high as about 6 dB. Then, the antenna under three different boundary conditions is deeply studied. The results reveal that the currents along the two sides of the metal ground are parallel to the slot radiator, resulting to affect the back-lobe radiation level to a great degree. As such, a pair of rectangular slots is properly etched on the two sides of the metal ground for further reduction of back-lobe radiation level and FBR improvement. Finally, a prototype antenna operating at 4.7 GHz is designed and fabricated. The measured results demonstrate that the back-lobe is indeed greatly decreased as predicted and the measured FBR of the proposed slot antenna exceeds 25 dB, where an increment of about 19 dB than the conventional λ/4 slot counterpart is successfully obtained, thereby evidently exhibiting better end-fire radiation performance.

Emerging building analytics heavily rely on data-driven machine learning algorithms. However, writing these analytics is still challenging: developers not only need to know what data is required but also where this data is in each individual building when writing applications. To bridge this gap between analytics and the actual resources in buildings, we present EnergonQL, a building independent acquisitional data query language that extracts data for building analytics with a declarative query processor. EnergonQL provides logic views of building resources that universally apply to all buildings, thus allowing portable building analytics across buildings. We evaluate EnergonQL with four different building analytics and show that with EnergonQL the line-of-code and development efforts can be effectively reduced.

In this paper, a differential-fed dual-band slot antenna with end-fire radiation is proposed. The dual-band characteristic is realized with help of three resonant modes of the λ/4 slot antenna, namely, the first odd mode in the lower band and the second and third odd-mode for the higher band. By symmetrically introducing two slot stubs around the nodal lines of electric-field distributions of the third odd mode, its resonant frequency gradually moves close to that of the second odd-mode. Thus, the bandwidth of the upper operating band could be significantly improved under operation of the second and third odd modes. To validate these predicted performances, a prototype antenna is in final fabricated and tested. The operating bandwidth of the antenna is about 4% in the lower band and 24% in the upper band. Both the simulated and measured results are provided with good agreement.

Purpose The present study aimed to investigate and compare the psychometric properties of the National Eye Institute Visual Functioning Questionnaire‐25 (NEI VFQ‐25) and the Visual Function Index‐14 (VF‐14) in a large sample of patients with cataracts. Methods A total of 1052 patients with bilateral age‐related cataracts were recruited in the study. Patients with other comorbidities that severely impacted vision were excluded. Participants completed the two questionnaires in random order. Classical test theory and Rasch analyses were used to assess the psychometric properties of the questionnaires. Results Complete data were obtained from 899 patients. The mean overall index score on the NEI VFQ‐25 was 76.1 ± 19.0, while that on the VF‐14 was 46.5 ± 15.0. Cronbach's α‐values for the NEI VFQ‐25 and VF‐14 were 0.89 and 0.95, respectively. Ceiling effects were observed on nine of the 12 subscales in the NEI VFQ‐25. The correlation between total scores on the NEI VFQ‐25 and VF‐14 was moderate (r = 0.600; p

RACK7 recognizes H3.3G34R to suppress expression of MHC II complex components and their delivery pathway in pediatric GBM., Histone H3 point mutations have been identified in incurable pediatric brain cancers, but the mechanisms through which these mutations drive tumorigenesis are incompletely understood. Here, we provide evidence that RACK7 (ZMYND8) recognizes the histone H3.3 patient mutation (H3.3G34R) in vitro and in vivo. We show that RACK7 binding to H3.3G34R suppresses transcription of CIITA, which is the master regulator of MHC (major histocompatibility complex) class II molecules and genes involved in vesicular transport of MHC class II molecules to the cell surface, resulting in suppression of MHC class II molecule expression and transport. CRISPR-based knock-in correction of the H3.3G34R mutation in human pediatric glioblastoma (pGBM) cells significantly reduces overall RACK7 chromatin binding and derepresses the same set of genes as does knocking out RACK7 in the H3.3G34R pGBM cells. By demonstrating that H3.3G34R and RACK7 work together, our findings suggest a potential molecular mechanism by which H3.3G34R promotes cancer.

Assembling for transcription initiation Eukaryotic transcription initiation by RNA polymerase II (Pol II) requires the assembly of a preinitiation complex (PIC) on core promoters. The binding of TATA box–binding protein (TBP) to the TATA box promoter has been thought to be a general rule in PIC assembly and transcription initiation. However, most coding genes lack a TATA box, and nearly all Pol II–mediated gene transcription requires the TBP-containing multisubunit complex transcription factor IID (TFIID). Chen et al. determined the structures of human TFIID-based PIC in sequential assembly states and revealed that TFIID supports distinct PIC assembly on TATA-containing and TATA-lacking promoters. The finding resolves the long-standing mystery of how one set of general transcription machinery initiates transcription on diverse promoters. Science , this issue p. eaba8490

Summary: Evolutionarily conserved SCAN (named after SRE-ZBP, CTfin51, AW-1, and Number 18 cDNA)-domain-containing zinc finger transcription factors (ZSCAN) have been found in both mouse and human genomes. Zscan4 is transiently expressed during zygotic genome activation (ZGA) in preimplantation embryos and induced pluripotent stem cell (iPSC) reprogramming. However, little is known about the mechanism of Zscan4 underlying these processes of cell fate control. Here, we show that Zscan4f, a representative of ZSCAN proteins, is able to recruit Tet2 through its SCAN domain. The Zscan4f-Tet2 interaction promotes DNA demethylation and regulates the expression of target genes, particularly those encoding glycolytic enzymes and proteasome subunits. Zscan4f regulates metabolic rewiring, enhances proteasome function, and ultimately promotes iPSC generation. These results identify Zscan4f as an important partner of Tet2 in regulating target genes and promoting iPSC generation and suggest a possible and common mechanism shared by SCAN family transcription factors to recruit ten-eleven translocation (TET) DNA dioxygenases to regulate diverse cellular processes, including reprogramming.

Tuberous sclerosis complex (TSC) integrates upstream stimuli and regulates cell growth by controlling the activity of mTORC1. TSC complex functions as a GTPase-activating protein (GAP) towards small GTPase Rheb and inhibits Rheb-mediated activation of mTORC1. Mutations in TSC genes cause tuberous sclerosis. In this study, the near-atomic resolution structure of human TSC complex reveals an arch-shaped architecture, with a 2:2:1 stoichiometry of TSC1, TSC2, and TBC1D7. This asymmetric complex consists of two interweaved TSC1 coiled-coil and one TBC1D7 that spans over the tail-to-tail TSC2 dimer. The two TSC2 GAP domains are symmetrically cradled within the core module formed by TSC2 dimerization domain and central coiled-coil of TSC1. Structural and biochemical analyses reveal TSC2 GAP-Rheb complimentary interactions and suggest a catalytic mechanism, by which an asparagine thumb (N1643) stabilizes γ-phosphate of GTP and accelerate GTP hydrolysis of Rheb. Our study reveals mechanisms of TSC complex assembly and GAP activity., Tuberous sclerosis complex (TSC) regulates cell growth by controlling the activity of mTORC1. The structure of human TSC complex reveals an arch-shaped, asymmetric architecture and a 2:2:1 stoichiometry of TSC1, TSC2, and TBC1D7 subunits and suggests a mechanism by which TSC2 accelerates GTP hydrolysis against a small GTPase Rheb.

Biliary atresia (BA) is a severe cholangiopathy that leads to liver failure in infants, but its pathogenesis remains to be fully characterized. By single-cell RNA profiling, we observed macrophage hypo-inflammation, Kupffer cell scavenger function defects, cytotoxic T cell expansion, and deficiency of CX3CR1+effector T and natural killer (NK) cells in infants with BA. More importantly, we discovered that hepatic B cell lymphopoiesis did not cease after birth and that tolerance defects contributed to immunoglobulin G (IgG)-autoantibody accumulation in BA. In a rhesus-rotavirus induced BA model, depleting B cells or blocking antigen presentation ameliorated liver damage. In a pilot clinical study, we demonstrated that rituximab was effective in depleting hepatic B cells and restoring the functions of macrophages, Kupffer cells, and T cells to levels comparable to those of control subjects. In summary, our comprehensive immune profiling in infants with BA had educed that B-cell-modifying therapies may alleviate liver pathology.

Dephosphorylating RNA polymerase II Transcription in metazoans requires coordination of multiple factors to control the progression of polymerases and the integrity of their RNA products. Zheng et al. identified a new dual-enzyme complex called INTAC, which is composed of protein phosphatase 2A (PP2A) core enzyme and the multisubunit RNA endonuclease Integrator. Structural and functional studies show that INTAC functions as a noncanonical PP2A holoenzyme that dephosphorylates the C-terminal domain of RNA polymerase II to attenuate transcription. This study provides a direct connection between PP2A-mediated dephosphorylation and transcriptional regulation, two fundamental cellular processes. Science , this issue p. eabb5872

The human coronavirus HCoV-19 infection can cause acute respiratory distress syndrome (ARDS), hypercoagulability, hypertension, extrapulmonary multiorgan dysfunction. Effective antiviral and anti-coagulation agents with safe clinical profiles are urgently needed to improve the overall prognosis. We screened an FDA approved drug library and found that an anticoagulant agent dipyridamole (DIP) suppressed HCoV-19 replication at an EC50 of 100 nMin vitro. It also elicited potent type I interferon responses and ameliorated lung pathology in a viral pneumonia model. In analysis of twelve HCoV-19 infected patients with prophylactic anti-coagulation therapy, we found that DIP supplementation was associated with significantly increased platelet and lymphocyte counts and decreased D-dimer levels in comparison to control patients. Two weeks after initiation of DIP treatment, 3 of the 6 severe cases (60%) and all 4 of the mild cases (100%) were discharged from the hospital. One critically ill patient with extremely high levels of D-dimer and lymphopenia at the time of receiving DIP passed away. All other patients were in clinical remission. In summary, HCoV-19 infected patients could potentially benefit from DIP adjunctive therapy by reducing viral replication, suppressing hypercoagulability and enhancing immune recovery. Larger scale clinical trials of DIP are needed to validate these therapeutic effects.

Post-translational modifications play vital roles in fine-tuning a myriad of physiological processes, and one of the most important modifications is acetylation. Here, we report a ligand-directed site-selective acetylation using KHAc, a derivative of a phosphoglycerate mutase 1 (PGAM1) inhibitor. KHAc binds to PGAM1 and transfers its acetyl group to the e-NH2 of Lys100 to inactivate the enzyme. The acetyl transfer process was visualized by time-resolved crystallography, demonstrating that the transfer is driven by proximity effects. KHAc was capable of selectively and effectively acetylating Lys100 of PGAM1 in cultured human cells, accompanied by inhibited F-actin formation. Similar strategies could be used for exogenous control of other lysine post-translational modifications.

Respiratory disease caused by the 2019 novel coronavirus (2019-nCoV) pneumonia first emerged in Wuhan, Hubei Province, China, in December 2019 and spread rapidly to other provinces and other countries. Angiotensin-converting enzyme 2 (ACE2) is the receptor for SARS-CoV and has been suggested to be also the receptor for 2019-nCoV. Paradoxically, ACE2 expression in the lung protects mice from SARS-CoV spike protein induced lung injury by attenuating the renin-angiotensin system. In the intestine, ACE2 also suppresses intestinal inflammation by maintaining amino acid homeostasis, antimicrobial peptide expression and ecology of the gut microbiome. Upon analysis of single cell-RNA sequencing data from control subjects and those with colitis or inflammatory bowel disease (IBD), we found thatACE2expression in the colonocytes was positively associated with genes regulating viral infection, innate and cellular immunity, but was negatively associated with viral transcription, protein translation, humoral immunity, phagocytosis and complement activation. In summary, we suggest that ACE2 may play dual roles in mediating the susceptibility and immunity of 2019-nCoV infection.