shRNA‑mediated knockdown of KNTC1 inhibits non-small-cell lung cancer through regulating PSMB8

In view of the important roles played by Kinetochore proteins in mitosis, we believed that they may contribute to the development and progression of human cancers, which has been reported recently elsewhere. Kinetochore-associated 1 (KNTC1) participates in the segregation of sister chromatids during mitosis, the effects of which on non-small-cell lung cancer (NSCLC) remain unclear. Here, we sought to identify the biological significance of KNTC1 in NSCLC. KNTC1 protein expression in NSCLC tissues was investigated by immunohistochemistry. Lentivirus delivered short hairpin RNA (shRNA) was utilized to establish KNTC1 silence NSCLC cell lines. The effects of KNTC1 depletion on NSCLC cell proliferation, migration, apoptosis, and tumor formation were analyzed by MTT assay, wound-healing assay, transwell assay, flow cytometry assay, and in nude mouse models in vivo. After KNTC1 reduction, NSCLC cell viability, proliferation, migration, and invasion were restrained. A xenograft tumor model was also provided to demonstrate the inhibited tumorigenesis in NSCLC. In addition, the downstream mechanism analysis indicated that KNTC1 depletion was positively associated with PSMB8. The findings of the present study suggested that KNTC1 may have a pivotal role in mediating NSCLC progression and may act as a novel therapeutic target for NSCLC.Subject terms: Non-small-cell lung cancer, Cell migration     

胶州湾典型河口湿地土壤活性有机碳和酶活性对互花米草入侵的响应

以胶州湾洋河口湿地为研究对象,按照互花米草入侵年份(0、1、5、8年)分层采集土壤样品(0—10、10—20、20—40 cm和40—60 cm),研究土壤活性有机碳(LOC)和酶(脲酶、过氧化氢酶、碱性磷酸酶、蔗糖酶)活性的变化,分析土壤活性有机碳和酶活性及两者相关性对互花米草入侵的响应。结果表明:与光滩相比,互花米草入侵增加了表层土壤LOC含量,且随着入侵时间的延长显著增加(P0.05)。同时也改变了土壤LOC垂直分布规律,除光滩和入侵1年样地表现出沿剖面逐渐上升之外,其他样地表现为沿剖面先上升后下降趋势;互花米草入侵提高了河口湿地土壤酶活性,但并未改变酶活性随深度增加而逐渐降低的分布规律。随着入侵时间的延长4种酶活性变化趋势有所差异,过氧化氢酶和蔗糖酶活性变化趋势一致,表现为随入侵时间延长先急剧增加后逐渐减少,而碱性磷酸酶和脲酶活性随着入侵时间的延长逐渐增加。Pearson相关性分析结果显示,土壤LOC和酶活性呈显著负相关且互花米草入侵时间越长两者间相关性越低,8年后无显著相关性。     

植被恢复过程中芒萁覆盖对侵蚀红壤氮组分的影响

氮素是限制陆地生态系统生产力的重要因子。采用时空代换法,以红壤侵蚀区未治理、恢复12年和30年的马尾松林为研究对象,对比分析了林下芒萁覆盖地与裸地表层土壤之间氮同位素、不同形态氮组分含量以及不同组分氮含量所占比例之间的差异。结果表明:在所有马尾松林中,芒萁覆盖增加了表层土壤的全氮含量,δ~(15)N值则比林下裸地显著降低了33. 8%—83.1%(P0.05)。随着恢复年限增加,林下芒萁覆盖地表层土壤δ~(15)N值显著下降,而林下裸露地δ~(15)N值没有显著变化(P0.05)。不同恢复年限马尾松林的芒萁覆盖地表层土壤微生物生物量氮、可溶性有机氮和铵态氮含量显著高于林下裸地(P 0.05),而硝态氮含量则显著低于林下裸地(P0.05)。随恢复年限增加,表层土壤微生物生物量氮、可溶性有机氮、铵态氮含量均呈增加趋势,而硝态氮含量则呈下降趋势,不同形态氮占全氮比例表现为:微生物生物量氮铵态氮可溶性有机氮硝态氮。相关分析表明土壤δ~(15)N值与硝态氮极显著正相关,与其他氮组分极显著负相关(P0.01)。由此可见,与林下裸地相比,芒萁覆盖在植被恢复过程中有助于提高表层土壤中全氮、微生物生物量氮、可溶性有机氮和铵态氮含量,降低硝态氮的淋溶损失风险,促进土壤氮保持和积累,从而有利于退化红壤生态系统的恢复。     

三种杓兰根相关真菌多样性和生态功能

【背景】除了菌根真菌(Orchid mycorrhizal fungi,OrMF)外,兰科植物根中还有其它内生真菌,称为根相关真菌(Root-associated fungi,RAF)。【目的】采用分离培养的方法获得同一栖息地针叶林和灌木林两种不同生境西藏杓兰、黄花杓兰和无苞杓兰的RAF菌株,研究其真菌谱系、多样性和生态功能结构。【方法】从杓兰根碎屑中分离RAF,通过总DNA提取、PCR扩增及测序得到ITS(Internaltranscribedspacer)序列;进行系统发育和多样性分析,并通过NCBI数据库比对得到相似性最高序列的注释信息来分析RAF生态学特性。【结果】共分离得到278株RAF,25种OTU类型,包括23个子囊菌门OTU,2个毛霉菌门OTU。RAF物种丰富度分析发现西藏杓兰的较黄花杓兰高,不同生境没有显著差异;不同杓兰物种较不同生境的RAF群落分化程度高。生态功能分析显示25个OTU包括共生型、腐生型和致病型3种营养型,以及外生菌根菌群、植物病原菌群、内生真菌群、动物病原菌群、真菌寄生菌群、杜鹃花类菌根群、未定义的腐生菌群和不确定型8种共位群。【结论】阐明不同生境采集的不同杓兰中RAF的分布特点和生态功能,为未来研究RAF与杓兰属植物的共生关系奠定基础。     

新疆棉花黄萎病菌拮抗细菌的分离、筛选与鉴定

【背景】棉花黄萎病是由大丽轮枝菌(Verticillium dahliae Kleb.)引起的一种世界性病害,近年来对该病害的生物防治因具有环境友好和人畜安全的特性而倍受关注。【目的】筛选棉花黄萎病高效拮抗细菌并对其进行鉴定,为棉花黄萎病的生物防治扩充菌种资源。【方法】采用稀释涂布平板法分离细菌,并进行拮抗细菌的初筛和复筛,通过形态特征、生理生化特征和16S rRNA基因序列分析对筛选到的细菌进行鉴定,确定其分类地位。【结果】初筛分离到535株对病原菌具有拮抗作用的细菌,并选取了108株拮抗细菌进行复筛,最终筛选到了4株优势拮抗细菌。通过形态观察、生理生化特征和16SrRNA基因序列分析,将菌株BHZ-29、SHT-15、SHZ-24和SMT-24分别鉴定为贝莱斯芽孢杆菌(Bacillusvelezensis)、枯草芽孢杆菌斯皮兹仁亚种(Bacillus subtilis subsp. spizizenii)、萎缩芽孢杆菌(Bacillus atrophaeus)和香草芽孢杆菌(Bacillus vanillea)。【结论】获得了4株高效拮抗细菌,并且首次报道了香草芽孢杆菌对棉花黄萎病菌具有抑制作用。     

Quantitative regulation of the thermal stability of enveloped virus vaccines by surface charge engineering to prevent the self-aggregation of attachment glycoproteins

The development of thermostable vaccines can relieve the bottleneck of existing vaccines caused by thermal instability and subsequent poor efficacy, which is one of the predominant reasons for the millions of deaths caused by vaccine-preventable diseases. Research into the mechanism of viral thermostability may provide strategies for developing thermostable vaccines. Using Newcastle disease virus (NDV) as model, we identified the negative surface charge of attachment glycoprotein as a novel determinant of viral thermostability. It prevented the temperature-induced aggregation of glycoprotein and subsequent detachment from virion surface. Then structural stability of virion surface was improved and virus could bind to and infect cells efficiently after heat-treatment. Employing the approach of surface charge engineering, thermal stability of NDV and influenza A virus (IAV) vaccines was successfully improved. The increase in the level of vaccine thermal stability was determined by the value-added in the negative surface charge of the attachment glycoprotein. The engineered live and inactivated vaccines could be used efficiently after storage at 37°C for at least 10 and 60 days, respectively. Thus, our results revealed a novel surface-charge-mediated link between HN protein and NDV thermostability, which could be used to design thermal stable NDV and IAV vaccines rationally.     

SARS-CoV-2-specific T cells associate with inflammation and reduced lung function in pulmonary post-acute sequalae of SARS-CoV-2

As of January 2022, at least 60 million individuals are estimated to develop post-acute sequelae of SARS-CoV-2 (PASC) after infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). While elevated levels of SARS-CoV-2-specific T cells have been observed in non-specific PASC, little is known about their impact on pulmonary function which is compromised in the majority of these individuals. This study compares frequencies of SARS-CoV-2-specific T cells and inflammatory markers with lung function in participants with pulmonary PASC and resolved COVID-19 (RC). Compared to RC, participants with respiratory PASC had between 6- and 105-fold higher frequencies of IFN-γ- and TNF-α-producing SARS-CoV-2-specific CD4⁺ and CD8⁺ T cells in peripheral blood, and elevated levels of plasma CRP and IL-6. Importantly, in PASC participants the frequency of TNF-α-producing SARS-CoV-2-specific CD4⁺ and CD8⁺ T cells, which exhibited the highest levels of Ki67 indicating they were activity dividing, correlated positively with plasma IL-6 and negatively with measures of lung function, including forced expiratory volume in one second (FEV¹), while increased frequencies of IFN-γ-producing SARS-CoV-2-specific T cells associated with prolonged dyspnea. Statistical analyses stratified by age, number of comorbidities and hospitalization status demonstrated that none of these factors affect differences in the frequency of SARS-CoV-2 T cells and plasma IL-6 levels measured between PASC and RC cohorts. Taken together, these findings demonstrate elevated frequencies of SARS-CoV-2-specific T cells in individuals with pulmonary PASC are associated with increased systemic inflammation and decreased lung function, suggesting that SARS-CoV-2-specific T cells contribute to lingering pulmonary symptoms. These findings also provide mechanistic insight on the pathophysiology of PASC that can inform development of potential treatments to reduce symptom burden.     

研究性实验教学模式的探讨

从教学过程、教学效果等方面探讨了微生物学实验课程实施研究性实验教学模式的必要性以及具体实施的方法。实践表明,研究性实验教学模式对培养学生的创新意识,提高学生的实践能力等方面有积极的作用。