Cloning and characterization of two ferritin subunit genes from bay scallop, <Emphasis Type="Italic">Argopecten irradians</Emphasis> (Lamarck 1819)

We have cloned two full-length cDNAs from two ferritin genes (Aifer1 and Aifer2) of the bay scallop, Argopecten irradians (Lamarck 1819). The cDNAs are 1,019 and 827 bp in length and encode proteins of 171 and 173 amino acids, respectively. The 5′ UTR of each contains a conserved iron response element (IRE) motif. Sequence analyses reveal that both proteins belong to the H-ferritin family with seven conserved amino acids in the ferroxidase center. Highest expression of Aifer1 is found in the mantle and adductor muscle, while that of Aifer2 is only in the latter tissue. These Aifer genes are differentially expressed following bacterial challenge of the scallop. The expression level of Aifer1 was acutely up-regulated (over 10 fold) at 6 h post-bacteria injection, whereas Aifer2 expression was not significantly changed by bacterial challenge. Both genes were effectively expressed in E. coli BL21 (DE3), producing proteins of similar molecular weight, approximately 23 kDa. Purified Aifer1 and Aifer2 proteins exhibited iron-chelating activity of 33.1% and 30.4%, respectively, at a concentration of 5 mg/ml. Cations, Mg²⁺, Zn²⁺ and Ca²⁺, depressed iron-chelating activity of both proteins. Additionally, the E. coli cells expressing recombinant Aifer1 and Aifer2 showed tolerance to H₂O₂, providing a direct evidence of the antioxidation function of ferritin. The results presented in this study suggest important roles of Aifer1 and Aifer2 in the regulation of iron homeostasis, immune response, and antioxidative stress in A. irradians.     

Effect of ubiquitin carboxy-terminal hydrolase 37 on apoptotic in A549 cells

Proteins destined for degradation by the ubiquitin-proteasome system are labelled with a 76-amino acid peptide, ubiquitin, through a series of conjugation steps by the E1, E2 and E3 enzymes respectively. Ubiquitin carboxy-terminal hydrolase 37 (UCH37) belongs to the UCH proteases family that deubiquitinates ubiquitin-protein conjugates in the ubiquitin-proteasome system. However, it is few reports about the relationship between UCH37 and apoptosis. In order to clarify the role of UCH37 on apoptosis, the A549 cells were chosen for this study. We transfected UCH37 siRNA and pcDNA3.1-UCH37 plasmid into A549 cells, respectively. Using MTT assay, Western blot, Hoechst 33342 staining assay and flow cytometry, we found that silencing of UCH37 in A549 cells induced apoptosis. The ratio of Bax/Bcl-2 was higher in silencing of UCH37 than that in control group after silencing of UCH37 in A549 cells. Meanwhile, experiments with the A549 cell line disclose that silencing of UCH37 could induce efficiently A549 cell apoptosis through activation of caspase-9 and caspase-3. On the other hand, over-expression of UCH37 led to the opposite effect. Hence, UCH37 might play an important role in apoptotic through altering Bax/Bcl-2 ratio and enzymatic activities of caspase-9 and caspase-3.     

苏州地区金线蛙食性的初步研究

金线蛙各月份的食性分析表明,成蛙以捕食昆虫为主,首食种类是水生动物中的负子昃、螺、虾等,摄食种类变化主要与生境及生长发育相关。     

Recombinant human interferon-α1b inhibits SARS-CoV-2 better than interferon-α2b in vitro

Highlights
1) A comprehensive evaluation method for anti-SARS-CoV-2 drugs was established based on RT-qPCR, TCID₅₀ method, and immunofluorescence.
2) A significant antiviral effect of rHuIFN-α1b was shown with EC₅₀=0.12 IU/mL in Vero cells and EC₅₀=0.52 IU/mL in Calu-3 cells, which was better than rHuIFN-α2b (EC₅₀=0.25 IU/mL in Vero cells and EC₅₀=2.48 IU/mL in Calu-3 cells).
3) rHuIFN-α1b has a good potential in the application of anti-COVID-19 therapy.     

渭干河-库车河绿洲景观生态安全时空分异及格局优化

渭干河-库车河绿洲(以下简称渭-库绿洲)是我国西北干旱区典型的荒漠绿洲区,维护其生态安全,是实现绿洲可持续发展的重要保障。以1997年、2006年及2016年3期Landsat TM影像数据为主要数据来源,构建渭-库绿洲景观生态安全评价模型,对近20年来研究区的景观生态安全时空特征进行分析。利用最小阻力模型,以水域、林草地为生态源地,将生态安全水平、海拔和坡度作为阻力因子生成最小累计阻力面,划分生态功能区,识别生态廊道和生态节点,从点、线、面综合视角进行景观格局优化。结果表明:(1)1997—2016年渭-库绿洲生态安全区域面积呈波动变化,相对安全区域面积在不断增加,临界安全、敏感和风险区域面积呈减小趋势,景观生态安全度在空间分布上由高到低呈内向外扩展的态势。(2)研究区景观生态安全的Moran′s I值分别为0.6479、0.7049、0.6587,景观生态安全值的空间集聚性呈先增加后降低的趋势;局部空间自相关主要以高-高聚集和低-低聚集类型为主,呈现出"同质聚集、异质分离"的特点。(3)景观格局优化中选取的生态源地占绿洲总面积的12.76%,构建的绿洲生态廊道基本贯穿整个研究区,关键廊道连接了绝大多数的绿洲生态源地,辅助廊道是连接没有与关键廊道连接的源地之间的通道,识别的生态节点主要分布在绿洲生态廊道的薄弱环节处,共计36个。将划分的生态缓冲区、生态连通区、生态过渡区、生态边缘区5个功能区和生态源地、生态廊道及生态节点景观组分相结合,并提出优化建议,以期为维持及进一步改善该绿洲生态环境提供科学依据。     

The Fugu tyrp1 promoter directs specific GFP expression in zebrafish: tools to study the RPE and the neural crest-derived melanophores

In vertebrates, pigment cells account for a small percentage of the total cell population and they intermingle with other cell types. This makes it difficult to isolate them for analyzes of their functions in the context of development. To alleviate such difficulty, we generated two stable transgenic zebrafish lines (pt101 and pt102) that express green fluorescent protein (GFP) in melanophores under the control of the 1 kb Fugu tyrp1 promoter. In pt101, GFP is expressed in both retinal pigment epithelium (RPE) cells and the neural crest-derived melanophores (NCDM), whereas in pt102, GFP is predominately expressed in the NCDM. Our results indicate that the Fugu tyrp1 promoter can direct transgene expression in a cell-type-specific manner in zebrafish. In addition, our findings provide evidence supporting differential regulations of melanin-synthesizing genes in RPE cells and the NCDM in zebrafish. Utilizing the varying GFP expression levels in these fish, we have isolated melanophores via flow cytometry and revealed the capability of sorting the NCDM from RPE cells as well. Thus, these transgenic lines are useful tools to study melanophores in zebrafish.