三角帆蚌Hc-GSK3β基因鉴定及内壳色性状相关SNP筛选

为了探究三角帆蚌(Hyriopsis cumingii)糖原合成激酶-3β(GSK3β)基因对壳色的影响,研究采用RACE技术获得Hc-GSK3β基因cDNA全长1867 bp,其中包含1261 bp的ORF区编码420个氨基酸, ORF中含有一个S＿TKc结构域,该结构域序列高度保守。组织差异表达分析发现Hc-GSK3β基因在紫色蚌鳃、斧足、内脏团和边缘膜组织中表达量高于白色蚌的表达量(P<0.05),且在斧足和边缘膜表达差异水平达到极显著(P<0.01),而在紫色蚌闭壳肌组织中表达量显著低于白色蚌(P<0.05)。原位杂交(ISH)实验结果显示在三角帆蚌外套膜的外褶、中褶、內褶、背膜区和腹膜区均有阳性信号产生,且在外褶的信号表达较强烈。该基因经重测序比较,共鉴定出6个SNP位点,其中在C+185A位点的CA基因型在紫色蚌的分布频率显著高于白色三角帆蚌(P<0.05);在紫色蚌中, T+341G位点TT基因型三角帆蚌内壳颜色参数b值显著低于TG基因型(P<0.05)。研究表明, Hc-GSK3β基因参与了三角帆蚌壳色形成,筛选的SNP标记可用于三角帆蚌壳...     

Quantitative Detection of 15 Serum Bile Acid Metabolic Products by LC/MS/MS in the Diagnosis of Primary Biliary Cholangitis

To determine 15 bile acid metabolic products in human serum by liquid chromatography-tandem mass spectrometry (LC/MS/MS) and value their diagnostic outcome in primary biliary cholangitis (PBC). Serum from 20 healthy controls and 26 patients with PBC were collected and went LC/MS/MS analysis of 15 bile acid metabolic products. The test results were analyzed by bile acid metabolomics, and the potential biomarkers were screened and their diagnostic performance was judged by statistical methods such as principal component and partial least squares discriminant analysis and area under curve (AUC). 8 differential metabolites can be screened out: Deoxycholic acid (DCA), Glycine deoxycholic acid (GDCA), Lithocholic acid (LCA), Glycine ursodeoxycholic acid (GUDCA), Taurolithocholic acid (TLCA), Tauroursodeoxycholic acid (TUDCA), Taurodeoxycholic acid (TDCA), Glycine chenodeoxycholic acid (GCDCA). The performance of the biomarkers was evaluated by the AUC, specificity and sensitivity. In conclusion, DCA, GDCA, LCA, GUDCA, TLCA, TUDCA, TDCA and GCDCA were identified as eight potential biomarkers to distinguish between healthy people and PBC patients by multivariate statistical analysis, which provided reliable experimental basis for clinical practice.     

Positional cloning and characterization reveal the role of a miRNA precursor gene ZmLRT in the regulation of lateral root number and drought tolerance in maize

Lateral roots play essential roles in drought tolerance in maize(Zea mays L.). However, the genetic basis for the variation in the number of lateral roots in maize remains elusive. Here, we identified a major quantitative trait locus(QTL),q LRT5-1, controlling lateral root number using a recombinant inbred population from a cross between the maize lines Zong3(with many lateral roots) and 87-1(with few lateral roots).Fine-mapping and functional analysis determined that the candidate gene for qLRT...     

A distinctive latitudinal trend of nitrogen isotope signature across urban forests in eastern China

Rapid urbanization has greatly altered nitrogen (N) cycling from regional to global scales. Compared to natural forests, urban forests receive much more external N inputs with distinctive abundances of stable N isotope (δ¹⁵N). However, the large-scale pattern of soil δ¹⁵N and its imprint on plant δ¹⁵N remain less well understood in urban forests. By collecting topsoil (0–20 cm) and leaf samples from urban forest patches in nine large cities across a north–south transect in eastern China, we analyzed the latitudinal trends of topsoil C:N ratio and δ¹⁵N as well as the correlations between tree leaf δ¹⁵N and topsoil δ¹⁵N. We further explored the spatial variation of topsoil δ¹⁵N explained by corresponding climatic, edaphic, vegetation-associated, and anthropogenic drivers. Our results showed a significant increase of topsoil C:N ratio towards higher latitudes, suggesting lower N availability at higher latitudes. Topsoil δ¹⁵N also increased significantly at higher latitudes, being opposite to the latitudinal trend of soil N availability. The latitudinal trend of topsoil δ¹⁵N was mainly explained by mean annual temperature, mean annual precipitation, and atmospheric deposition of both ammonium and nitrate. Consequently, tree leaf δ¹⁵N showed significant positive correlations with topsoil δ¹⁵N across all sampled plant species and functional types. Our findings reveal a distinctive latitudinal trend of δ¹⁵N in urban forests and highlight an important role of anthropogenic N sources in shaping the large-scale pattern of urban forest ¹⁵N signature.     

猪链球菌2型Ⅳ型分泌系统组分VirD4与毒力相关性分析

摘要:【目的】构建猪链球菌2型强毒株05ZYH33毒力岛89 K上的Ⅳ型分泌系统组分VirD4敲除突变株,初步分析其活性和毒力,为进一步研究猪链球菌2 型在逃避宿主天然免疫杀伤中的作用提供基础。【方法】以05ZYH33基因组为模板,PCR扩增VirD4基因上下游同源臂,以穿梭质粒pSET1为模板,PCR扩增氯霉素抗性基因Cm,通过重叠PCR技术搭建上述3个片段并连接至温敏载体pSET4s,构建基因敲除载体pSET4s∷VirD4;通过同源重组构建基因敲除突变株ΔVirD4;通过体外全血杀伤实验、CD1小鼠竞争感染及攻毒实验 对突变株和野生株的毒力进行比较分析。【结果】获得了基因敲除突变株ΔVirD4,通过对比发现其毒力与野生株相比有所降低。【结论】猪链球菌2型Ⅳ型分泌系统组分VirD 与其毒力相关,并在早期抵抗天然免疫细胞杀伤中发挥一定作用。     

Ebastine exerts antitumor activity and induces autophagy by activating AMPK/ULK1 signaling in an IPMK-dependent manner in osteosarcoma

Numerous studies have confirmed that in addition to interfering with the tumor inflammatory environment, anti-inflammatory agents can directly increase apoptosis and sensitivity to conventional therapies and decrease invasion and metastasis, making them useful candidates for cancer therapy. Here, we first used high-throughput screening and had screened one compound candidate, ebastine (a H1-histamine receptor antagonist), for osteosarcoma therapy. Cell viability assays, colony formation assays, wound healing assays, and Transwell assays demonstrated that ebastine elicited antitumor effects in osteosarcoma cells. In addition, ebastine treatment exerted obvious effects on cell cycle arrest, metastasis inhibition, apoptosis and autophagy induction both in vitro and in vivo. Mechanistically, we observed that ebastine treatment triggered proapoptotic autophagy by activating AMPK/ULK1 signaling in osteosarcoma cells. Treatment with the AMPK inhibitor dorsomorphin reversed ebastine-induced apoptosis and autophagy. More importantly, we found that IPMK interacted with AMPK and functioned as a positive regulator of AMPK protein in osteosarcoma cells. A rescue study showed that the induction of autophagy and activation of the AMPK/ULK1 signaling pathway by ebastine treatment were reversed by IPMK knockdown, indicating that the activity of ebastine was IPMK dependent. We provide experimental evidence demonstrating that ebastine has antitumor activity in osteosarcoma and promotes autophagy by activating the AMPK/ULK1 signaling pathway, which is IPMK dependent. Our results provide insight into the clinical application potential of ebastine, which may represent a new potential therapeutic candidate for the treatment of osteosarcoma.     

Competition and abiotic stress affect the size of mangroves near their geographic range limit

竞争和非生物胁迫影响处于地理分布边界的红树植物的个体大小 关于红树植物竞争的研究大多局限于幼苗和人工林。我们首次对天然红树林中成年红树的种内竞争进行了控制实验研究,旨在检验竞争和非生物因子在决定红树植物个体大小中的相对重要性。研究样 地位于靠近红树林地理分布边界的美国德克萨斯州阿兰萨斯港(Port Aransas)附近区域。该区域的红树林由“灌丛”状的黑红树(萌芽白骨壤,Avicennia germinans)单一物种组成。我们对10个样方中原生红树 林进行疏伐,形成系列红树林覆盖度梯度,在2013–2019年期间观测各样方中红树植物的生长指标,量化分析红树林覆盖度对红树植物生长的影响;并于2019年调查了红树林的冠层高度。研究结果表明,在该研究期间,红树植物的相对生长速率随着红树林覆盖度的增加而降低,100%红树林覆盖度样方中的红 树植物大小几乎没有增长,说明它们已经达到了该红树林密度条件下的最大尺寸。在红树林覆盖度降低 的样方中,株高明显增加,在红树林覆盖度为11%的样方中,红树植物株高增加了约52%。对比临水岸 边和林内两种生境中的样方,处于临水岸边生境的红树林冠层高度比处于林内生境的高约30%,且这两 种生境的红树林冠层高度均随红树林覆盖度的增加而降低。叶片叶绿素含量和冠层光截留量的测定数据 显示,该区域红树植物的生长也受到氮限制的影响。由此表明,处于地理分布边界的“灌丛”状红树林一 方面受到营养的限制,另一方面红树植物种内个体间仍存在较为强烈的竞争,且种内竞争对红树植物生长的影响较该红树林内非生物生境因子更为重要。     

Accumulation of Hydrocarbons by Maize (Zea mays L.) in Remediation of Soils Contaminated with Crude Oil

This study has investigated the use of screened maize for remediation of soil contaminated with crude oil. Pots experiment was carried out for 60 days by transplanting maize seedlings into spiked soils. The results showed that certain amount of crude oil in soil (≤2 147 mg·kg^?1) could enhance the production of shoot biomass of maize. Higher concentration (6 373 mg·kg^?1) did not significantly inhibit the growth of plant maize (including shoot and root). Analysis of plant shoot by GC-MS showed that low molecular weight polycyclic aromatic hydrocarbons (PAHs) were detected in maize tissues, but PAHs concentration in the plant did not increase with higher concentration of crude oil in soil. The reduction of total petroleum hydrocarbon in planted soil was up to 52.21–72.84%, while that of the corresponding controls was only 25.85–34.22% in two months. In addition, data from physiological and biochemical indexes demonstrated a favorable adaptability of maize to crude oil pollution stress. This study suggested that the use of maize (Zea mays L.) was a good choice for remediation of soil contaminated with petroleum within a certain range of concentrations.     

Highly sensitive and selective uric acid biosensor based on a three-dimensional graphene foam/indium tin oxide glass electrode

A three-dimensional (3D) continuous and interconnected network graphene foam (GF) was synthesized by chemical vapor deposition using nickel foam as a template. The morphologies of the GF were observed by scanning electron microscopy. X-ray diffraction and Raman spectroscopy were used to investigate the structure of GF. The graphene with few layers and defect free was closely coated on the backbone of the 3D nickel foam. After etching nickel, the GF was transferred onto indium tin oxide (ITO) glass, which acted as an electrode to detect uric acid using cyclic voltammetry (CV) and differential pulse voltammetry (DPV). The GF/ITO electrode showed a high sensitivity for the detection of uric acid: approximately 9.44 mA mM⁻¹ in the range of 25 nM–0.1 μM and 1.85 mA mM⁻¹ in the range of 0.1–60 μM. The limit of detection of GF/ITO electrode for uric acid is 3 nM. The GF/ITO electrode also showed a high selectivity for the detection of uric acid in the presence of ascorbic acid. This electrode will have a wide range of potential application prospects in electrochemical detection.     

Comparative Genomic Analysis of 45 Type Strains of the Genus Bifidobacterium: A Snapshot of Its Genetic Diversity and Evolution

Bifidobacteria are well known for their human health-promoting effects and are therefore widely applied in the food industry. Members of the Bifidobacterium genus were first identified from the human gastrointestinal tract and were then found to be widely distributed across various ecological niches. Although the genetic diversity of Bifidobacterium has been determined based on several marker genes or a few genomes, the global diversity and evolution scenario for the entire genus remain unresolved. The present study comparatively analyzed the genomes of 45 type strains. We built a robust genealogy for Bifidobacterium based on 402 core genes and defined its root according to the phylogeny of the tree of bacteria. Our results support that all human isolates are of younger lineages, and although species isolated from bees dominate the more ancient lineages, the bee was not necessarily the original host for bifidobacteria. Moreover, the species isolated from different hosts are enriched with specific gene sets, suggesting host-specific adaptation. Notably, bee-specific genes are strongly associated with respiratory metabolism and are potential in helping those bacteria adapt to the oxygen-rich gut environment in bees. This study provides a snapshot of the genetic diversity and evolution of Bifidobacterium, paving the way for future studies on the taxonomy and functional genomics of the genus.