Cervical alignment was compromised after laminoplasty in patients with CSM, and the degree of LCL was associated with preoperative T1 slope, C2-7 SVA, and CVLL.

Dear Editor, Since the outbreak of a novel coronavirus disease (COVID-19) in late 2019, it has spread rapidly and developed into a global pandemic. As of August 12, 2020, more than 215 countries and territories around the world have reported more than 20.5 million confirmed COVID-19 cases with over 745,693 deaths (https://www. worldometers.info/coronavirus/#countries). Such harsh conditions urged scientists across the world to gear up to develop vaccines and antiviral drugs against COVID-19, which also lead to massive requirement for experimental animals. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative pathogen of COVID-19. It has been demonstrated that SARS-CoV-2 uses angiotensin converting enzyme 2 (ACE2) as cellular receptor for entry into target cells. Mouse model is the most commonly used animal model for studying human diseases. However, SARS-CoV-2 fails to invade and replicate in this traditional animal model due to the structural differences in mouse ACE2 (mACE2) compared with human ACE2 (hACE2), 1 which has become the major hurdle for COVID-19 study. Currently, several strategies have been developed to overcome this receptor incompatibility by: (i) generating transgenic mice bearing hACE2 receptor, 2-4 (ii) establishing adenovirus hACE2 mouse model with recombinant adenovirus expressing hACE2, 5 and (iii) adapting the SARS-CoV-2 by serial passages in the respiratory tract of mice. 6-8 In this study, we used an alternative strategy to generate a SARS-CoV-2-sensitive mouse model by exogenous delivery of hACE2 with Venezuelan equine encephalitis replicon particles (VEEV-VRP-hACE2) (Supplementary information, Fig. S1a). VEEV is a positive sense, single-stranded RNA virus which belongs to the genus Alphavirus, family Togaviridae. Alphavirus replicon particles (VRPs), including VEEV-VRPs, represent efficient vectors for gene delivery and have been applied to studies of vaccine development, gene therapy and cell transduction. They contain self-replicating RNAencoding viral replicase proteins (nsP1-nsP4) and express the gene of interest in place of viral structural protein genes. 9 By providing viral structural proteins in trans, the replicon RNA is packaged into VEEV-VRPs for in vitro and in vivo gene delivery. 10 Due to their intrinsic biological properties, VEEV-VRPs offer several advantages with a broad range of susceptible host cells, high expression level of cytoplasmic proteins and easy manipulation of recombinant RNA molecules using cDNA clones. 10,11 Here, Venezuelan equine encephalitis virus (VEEV) replicon expressing hACE2 with a C-terminal Stag was packaged into VRPs using the helper RNAs encoding VEEV capsid and envelope proteins to produce VEEV-VRP-hACE2 (Supplementary information, Fig. S1). MLE-12 cells (mouse lung type II epithelial cell line) were used to evaluate the availability of VEEV-VRP-hACE2 for SARS-CoV-2-sensitive cells establishment. After confirming hACE2 expression in MLE-12 cells transduced with VEEV-VRP-hACE2 (VRP-hACE2) through indirect immunofluoresc...

Because Iron (Fe) is an essential element, Fe storage in plant seeds is necessary for seedling establishment following germination. However, the mechanisms controlling seed Fe storage during seed development remain largely unknown. Here we reveal that an ERF95 transcription factor regulatesArabidopsisseed Fe accumulation. We show that expression ofERF95increases during seed maturation, and that lack of ERF95 reduces seed Fe accumulation, consequently increasing sensitivity to Fe deficiency during seedling establishment. Conversely, overexpression ofERF95has the opposite effects. We show that lack of ERF95 decreases abundance ofFER1messenger RNA in developing seed, which encodes Fe‐sequestering ferritin. Accordingly, afer1‐1loss‐of‐function mutation confers reduced seed Fe accumulation, and suppresses ERF95‐promoted seed Fe accumulation. In addition, ERF95 binds to specificFER1promoter GCC‐boxes and transactivatesFER1expression. We show thatERF95expression in maturing seed is dependent on EIN3, the master transcriptional regulator of ethylene signaling. While lack of EIN3 reduces seed Fe content, overexpression ofERF95rescues Fe accumulation in the seed ofein3loss‐of‐function mutant. Finally, we show that ethylene production increases during seed maturation. We conclude that ethylene promotes seed Fe accumulation during seed maturation via an EIN3‐ERF95‐FER1‐dependent signaling pathway.

The severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) virus, which is highly pathogenic and classified as a biosafety level 3 (BSL-3) agent, has greatly threatened global health and efficacious antivirals are urgently needed. The high requirement of facilities to manipulate the live virus has limited the development of antiviral study. Here, we constructed a reporter replicon of SARS-CoV-2, which can be handled in a BSL-2 laboratory. The Renilla luciferase activity effectively reflected the transcription and replication levels of the replicon genome. We identified the suitability of the replicon in antiviral screening using the known inhibitors, and thus established the replicon-based high-throughput screening (HTS) assay for SARS-CoV-2. The application of the HTS assay was further validated using a few hit natural compounds, which were screened out in a SARS-CoV-2 induced cytopathic-effect-based HTS assay in our previous study. This replicon-based HTS assay will be a safe platform for SARS-CoV-2 antiviral screening in a BSL-2 laboratory without the live virus.