Time-Series Clustering of lncRNA-mRNA Expression during the Adipogenic Transdifferentiation of Porcine Skeletal Muscle Satellite Cells

Skeletal muscle satellite cells (SMSCs), which are multifunctional muscle-derived stem cells, can differentiate into adipocytes. Long-chain non-coding RNA (lncRNA) has diverse biological functions, including the regulation of gene expression, chromosome silencing, and nuclear transport. However, the regulatory roles and mechanism of lncRNA during adipogenic transdifferentiation in muscle cells have not been thoroughly investigated. Here, porcine SMSCs were isolated, cultured, and induced for adipogenic differentiation. The expressions of lncRNA and mRNA at different time points during transdifferentiation were analysed using RNA-seq analysis. In total, 1005 lncRNAs and 7671 mRNAs showed significant changes in expression at differential differentiation stages. Time-series expression analysis showed that the differentially expressed (DE) lncRNAs and mRNAs were clustered into 5 and 11 different profiles with different changes, respectively. GO, KEGG, and REACTOME enrichment analyses revealed that DE mRNAs with increased expressions during the trans-differentiation were mainly enriched in the pathways for lipid metabolism and fat cell differentiation. The genes with decreased expressions were mainly enriched in the regulation of cell cycle and genetic information processing. In addition, 1883 DE mRNAs were regulated by 193 DE lncRNAs, and these genes were related to the controlling in cell cycle mainly. Notably, three genes in the fatty acid binding protein (FABP) family significantly and continuously increased during trans-differentiation, and 15, 13, and 11 lncRNAs may target FABP3, FABP4, and FABP5 genes by cis- or trans-regulation, respectively. In conclusion, these studies identify a set of new potential regulator for adipogenesis and cell fate and help us in better understanding the molecular mechanisms of trans-differentiation.     

Validation Study to Determine the Accuracy of Widespread Promoterless EGFP Reporter at Assessing CRISPR/Cas9-Mediated Homology Directed Repair

An accurate visual reporter system to assess homology-directed repair (HDR) is a key prerequisite for evaluating the efficiency of Cas9-mediated precise gene editing. Herein, we tested the utility of the widespread promoterless EGFP reporter to assess the efficiency of CRISPR/Cas9-mediated homologous recombination by fluorescence expression. We firstly established a promoterless EGFP reporter donor targeting the porcine GAPDH locus to study CRISPR/Cas9-mediated homologous recombination in porcine cells. Curiously, EGFP was expressed at unexpectedly high levels from the promoterless donor in porcine cells, with or without Cas9/sgRNA. Even higher EGFP expression was detected in human cells and those of other species when the porcine donor was transfected alone. Therefore, EGFP could be expressed at certain level in various cells transfected with the promoterless EGFP reporter alone, making it a low-resolution reporter for measuring Cas9-mediated HDR events. In summary, the widespread promoterless EGFP reporter could not be an ideal measurement for HDR screening and there is an urgent need to develop a more reliable, high-resolution HDR screening system to better explore strategies of increasing the efficiency of Cas9-mediated HDR in mammalian cells.     

TEB/POLQ plays dual roles in protecting Arabidopsis from NO-induced DNA damage

Nitric oxide (NO) is a key player in numerous physiological processes. Excessive NO induces DNA damage, but how plants respond to this damage remains unclear. We screened and identified an Arabidopsis NO hypersensitive mutant and found it to be allelic to TEBICHI/POLQ, encoding DNA polymerase θ. The teb mutant plants were preferentially sensitive to NO- and its derivative peroxynitrite-induced DNA damage and subsequent double-strand breaks (DSBs). Inactivation of TEB caused the accumulation of spontaneous DSBs largely attributed to endogenous NO and was synergistic to DSB repair pathway mutations with respect to growth. These effects were manifested in the presence of NO-inducing agents and relieved by NO scavengers. NO induced G2/M cell cycle arrest in the teb mutant, indicative of stalled replication forks. Genetic analyses indicate that Polθ is required for translesion DNA synthesis across NO-induced lesions, but not oxidation-induced lesions. Whole-genome sequencing revealed that Polθ bypasses NO-induced base adducts in an error-free manner and generates mutations characteristic of Polθ-mediated end joining. Our experimental data collectively suggests that Polθ plays dual roles in protecting plants from NO-induced DNA damage. Since Polθ is conserved in higher eukaryotes, mammalian Polθ may also be required for balancing NO physiological signaling and genotoxicity.     

VEGF/VEGFR-Targeted Therapy and Immunotherapy in Non-small Cell Lung Cancer: Targeting the Tumor Microenvironment

Non-small cell lung cancer (NSCLC) is the leading cause of death by cancer worldwide. Despite developments in therapeutic approaches for the past few decades, the 5-year survival rate of patients with NSCLC remains low. NSCLC tumor is a complex, heterogeneous microenvironment, comprising blood vessels, cancer cells, immune cells, and stroma cells. Vascular endothelial growth factors (VEGFs) are a major mediator to induce tumor microvasculature and are associated with the progression, recurrence, and metastasis of NSCLC. Current treatment medicines targeting VEGF/VEGF receptor (VEGFR) pathway, including neutralizing antibodies to VEGF or VEGFR and receptor tyrosine kinase inhibitors, have shown good treatment efficacy in patients with NSCLC. VEGF is not only an important angiogenic factor but also an immunomodulator of tumor microenvironment (TME). VEGFs can suppress antigen presentation, stimulate activity of regulatory T (Treg) cells, and tumor-associated macrophages, which in turn promote an immune suppressive microenvironment in NSCLC. The present review focuses on the angiogenic and non-angiogenic functions of VEGF in NSCLC, especially the interaction between VEGF and the cellular components of the TME. Additionally, we discuss recent preclinical and clinical studies to explore VEGF/VEGFR-targeted compounds and immunotherapy as novel approaches targeting the TME for the treatment of NSCLC.     

鄱阳湖沙地蔓荆灌丛沙堆形态特征及空间分布格局

目前灌丛沙堆研究主要集中在干旱半干旱区的沙质草原和沙漠边缘,对亚热带湿润区灌丛沙堆的形成、演变过程并不清楚。以鄱阳湖沙地为研究区,通过样方调查和地统计学的方法,研究不同沙化程度下蔓荆灌丛沙堆的形态特征及分布格局。结果表明:鄱阳湖沙地蔓荆沙堆的形态以盾形为主,其冠幅变化幅度为1.2—18.2 m~2,固定和半固定沙地显著高于流动沙地;对灌丛沙堆的形态参数来说,其长轴与短轴在固定和半固定沙地上呈极显著的线性相关关系,流动沙地上呈二次函数关系;半固定和流动沙地上沙堆底面积与沙堆高度呈二次函数关系(r0.6);3种类型沙地上灌丛底面积与沙堆体积之间极显著线性相关,其中半固定沙地线性函数的斜率最大;除固定沙地的沙堆高度和半固定沙地的灌丛高度外,3种沙地上蔓荆灌丛与沙堆的其他形态参数间均极显著相关,说明随着沙地的固定,蔓荆灌丛有利于沙堆水平尺度的增长;3种沙地上蔓荆沙堆均呈随机分布。     

Assessing the impact of ambient temperature on the risk of hand,foot, and mouth disease in Guangdong,China: New insight from the disease severity and burden

BackgroundThe association between the incidence of hand, foot, and mouth disease (HFMD) and ambient temperature has been well documented. Although the severity of symptoms is an important indicator of disease burden and varies significantly across cases, it usually was ignored in previous studies, potentially leading to biased estimates of the health impact of temperature.MethodsWe estimated the disability-adjusted life year (DALY) by considering the severity of symptoms for each HFMD case reported during 2010–2012 in Guangdong and used distributed lag-nonlinear models to estimate the association between the daily average temperature and daily DALY of HFMD cases at the city-level. We investigated the potential effect modifiers on the pathway between temperature and DALY and pooled city-specific estimates to a provincial association using a meta-regression. The overall impact of temperature was further evaluated by estimates of DALYs that could be attributed to HFMD.ResultsThe overall cumulative effect of daily mean temperature on the DALY of HFMD showed an inverse-U shape, with the maximum effect estimated to be β = 0.0331 (95%CI: 0.0199–0.0463) DALY at 23.8°C. Overall, a total of 6.432 (95%CI: 3.942–8.885) DALYs (attributable fraction = 2.721%, 95%CI: 1.660–3.759%) could be attributed to temperature exposure. All the demographic subgroups had a similar trend as the main analysis, while the magnitude of the peak of the temperature impact tended to be higher among the males, those aged ≥3yrs or from the Pear-River Delta region. Additionally, the impact of temperature on DALY elevated significantly with the increasing population density, per capita GDP, and per capita green space in parks.ConclusionsTemperature exposure was associated with increased burden of HFMD nonlinearly, with certain groups such as boys and those from areas with greater population density being more vulnerable.