Studying bona fide SARS-CoV-2 biology in a BSL-2 biosafety environment using a split-virus-genome system

<正>Dear Editor,Severe acute respiratory syndrome coronavirus 2(SARSCoV-2) was identified as the pathogen causing the coronavirus disease(COVID-19), which sometimes resulted in fatal pneumonia(Hu et al., 2021). SARS-CoV-2 is a biosafety level 3(BSL-3) pathogen, and the requirement for high containment conditions is a bottleneck for basic research on viral biology.     

Reagentless amperometric immunosensor for human chorionic gonadotrophin based on direct electrochemistry of horseradish peroxidase

A novel amperometric immunosensor for determination of human serum chorionic gonadotrophin (HCG) was constructed by immobilization of HCG with titania sol-gel on a glassy carbon electrode and the direct electrochemistry of horseradish peroxidase (HRP) labeled to HCG antibody (HRP-anti-HCG). The morphologies of the HCG membrane were characterized to be chemically clean, porous and homogeneous. HRP-anti-HCG was functionally conjugated with the immobilized HCG after incubation in phosphate buffer (PBS) containing HRP-anti-HCG. A direct electron transfer of HRP with a rate constant of 1.35+/-0.40 s(-1) was observed at the HRP-anti-HCG-HCG/titania sol-gel membrane modified electrode in 0.1 M PBS pH 7.0. With a competitive mechanism the differential pulse voltammetric peak current of the immobilized HRP decreased linearly with an increasing HCG concentration from 2.5 to 12.5 mIU/ml in the incubation solution. The HCG immunosensor showed a detection limit of 1.4 mIU/ml, a good accuracy and acceptable precision and reproducibility with an intra-assay CV of 4.7% at 5.0 mIU/ml and an inter-assay precision of 8.1% obtained at 10 mIU/ml. The biosensor displayed a good stability in a storage period of 30 days.     

Biodegradation of poly(ε-caprolactone) (PCL) by a new Penicillium oxalicum strain DSYD05-1

In this study, fungi isolated from soil were screened for their ability to form clear zones on agar plates with emulsified poly(ε-caprolactone) (PCL). The most active strain, designated as DSYD05, was identified as Penicillium oxalicum on the basis of morphological characteristics and phylogenetic analysis. Mutant DSYD05-1, obtained by ultraviolet-light mutagenesis from strain DSYD05, was more effective in PCL degradation. In liquid cultures of the mutant strain with PCL emulsion, DSYD05-1 showed the highest PCL-degrading activity after 4?days of cultivation. The products of PCL degradation were analysed by mass spectrometry; the results indicated that 6-hydroxyhexanoic acid was produced and assimilated during cultivation. The degradation of PCL film by DSYD05-1 was observed by scanning electron microscopy, and was indicative of a three-stage degradation process. The degradation of amorphous parts of the film preceded that of the crystalline center and then the peripheral crystalline regions. In addition, DSYD05-1 showed a wide range of substrate specificity, with capability to degrade PCL, poly(β-hydroxybutyrate), and poly(butylene succinate), but not poly(lactic acid), indicating that the strain could have potential for application in the treatment or recycling of bio-plastic wastes.     

A novel spliced variant of the type 1 corticotropin-releasing hormone receptor with a deletion in the seventh transmembrane domain present in the human pregnant term myometrium and fetal membranes

CRH exerts its actions via activation of specific G protein-coupled receptors, which exist in two types, CRH-R1 and CRH-R2, and arise from different genes with multiple spliced variants. RT-PCR amplification of CRH receptor sequences from human myometrium and fetal membranes yielded cDNAs that encode a novel CRH-R type 1 spliced variant. This variant (CRH-R1d) is present in the human pregnant myometrium at term only, which suggests a physiologically important role at the end of human pregnancy and labor. The amino acid sequence of CRH-R1d is identical to the CRH-R1alpha receptor except that it contains an exon deletion resulting in the absence of 14 amino acids in the predicted seventh transmembrane domain. Binding studies in HEK-293 cells stably expressing the CRH-R1d or CRH-R1alpha receptors revealed that the deletion does not change the binding characteristics of the variant receptor. In contrast, studies on the G protein activation demonstrated that CRH-R1d is not well coupled to the four subtypes of G proteins (G(s), G(i), G(o), G(q)) that CRH-R1alpha can activate. These data suggest that although the deleted segment is not important for CRH binding, it plays a crucial role in CRH receptor signal transduction. Second messenger studies of the variant receptor showed that CRH and CRH-like peptides can stimulate the adenylate cyclase system, with reduced sensitivity and potency by 10-fold compared with the CRH-R1alpha. Furthermore, CRH failed to stimulate inositol trisphosphate production. Coexpression studies between the CRH-R1d or CRH-R1alpha showed that this receptor does not play a role as a dominant negative receptor for CRH.     

微核技术研究进展

微核试验作为检测染色体损伤最常用而有效的细胞遗传学检测方法之一,经济、简便、快速,在敏感性、特异性和准确性方面,与经典的染色体畸变分析方法基本相当。主要综述国内外微核检测技术的最新研究进展,尤其是微核的自动化检测技术。其中流式细胞仪自动化检测和激光扫描细胞仪自动化检测,以及微核试验高内涵筛选方法由于其特有的优势,应用和发展前景广阔。     

The extracellular domain of p185(c-neu) induces density-dependent inhibition of cell growth in malignant mesothelioma cells and reduces growth of mesothelioma in vivo

EGFR is involved in the density-dependent inhibition of cell growth, while coexpression of EGFR with erbB2 can render normal cells transformed. In this study, we have examined the effect of a species of p185 that contains the transmembrane domain and the extracellular domain of p185(c-neu), on growth properties of a human malignant mesothelioma cell line that coexpresses EGFR and erbB2. The ectodomain form of p185(c-neu) enhanced density-dependent inhibition of cell growth and we found that p21 induction appeared to be responsible for this inhibitory effect. Previously, the extracellular domain species was shown to suppress the transforming abilities of EGFR and p185(c-neu/erbB2) in a dominant-negative manner. The ability of this subdomain to affect tumor growth is significant, as it reduced in vivo tumor growth. Unexpectedly, we found that the domain did not abrogate all of EGFR functions. We noted that EGFR-induced density-dependent inhibition of cell growth was retained. Tyrosine kinase inhibitors of EGFR did not cause density-dependent inhibition of cell growth of malignant mesothelioma cells. Therefore, simultaneously inhibiting the malignant phenotype and inducing density-dependent inhibition of cell growth in malignant mesothelioma cells by the extracellular domain of p185(c-neu) may represent an important therapeutic advance.     

红松阔叶林倒木贮量动态的研究

在森林倒木研究的基础上探讨长白山红松阔叶林倒木贮量的动态,涉及红松阔叶林倒木分解及其贮量的动态规律。研究表明,倒木分解,除心腐木外,均由表及里进行;倒木分解速率在其它生态条件相同时因树种、直径和部位而异。红松阔叶林倒木贮量动态包括现有倒木贮量和倒木年输入量两个分解动态过程,现有倒木贮量在头100年其干重迅速减少,其中椴树比红松尤速,前者分解91%,后者为72%.林地倒木贮量动态与倒木年输入量分解动态相似,但前者在分解初期贮量增加较大,因为部分现有倒木未完全分解;100年后趋于一致,并恒定于16～17t·hm^-2,直至群落的顶极阶段结束.     

禽致病性大肠杆菌毒力基因多重PCR方法的建立和应用

【目的】建立禽致病性大肠杆菌(Avian pathogenic Escherichia coli,APEC)黏附相关基因、侵袭及毒素相关基因、抗血清存活相关基因及铁转运相关基因的多重PCR方法,实现禽致病性大肠杆菌毒力基因的简便、快速检测。【方法】根据GenBank公布的基因序列,设计合成18对特异性引物,通过条件优化,建立四组多重PCR体系,并通过模板倍比稀释检测各组多重PCR的灵敏性。利用多重PCR检测100株APEC毒力基因的分布,验证多重PCR方法的可行性。【结果】根据PCR扩增片段大小判定,上述四组多重PCR体系均能同时扩增出该组中的各个毒力基因,且灵敏度分别为:103CFU、103CFU、105CFU、105CFU细菌和1ng、1ng、10ng、10ng DNA。100株APEC的毒力因子检测结果显示,多重PCR和单基因PCR结果一致。【结论】建立的四组多重PCR方法能够简便、快速地检测禽致病性大肠杆菌的毒力基因,可用于毒力基因的鉴定以及流行病学调查。