Circular RNA circDVL1 inhibits clear cell renal cell carcinoma progression through the miR-412-3p/PCDH7 axis

Clear cell renal cell carcinoma (ccRCC) is a primary kidney cancer with high aggressive phenotype and extremely poor prognosis. Accumulating evidence suggests that circular RNAs (circRNAs) play pivotal roles in the occurrence and development of various human cancers. However, the expression, clinical significance and regulatory role of circRNAs in ccRCC remain largely unclear. Here we report that circDVL1 to be reduced in the serums and tissues from ccRCC patients, and to negatively correlate with ccRCC malignant features. Overexpression of circDVL1 inhibits proliferation, induces G1/S arrest, triggers apoptosis, and reduces migration and invasion in different ccRCC cells in vitro. Correspondingly, circDVL1 overexpression suppresses ccRCC tumorigenicity in a mouse xenograft model. Mechanistically, circDVL1 serves as a sponge for oncogenic miR-412-3p, thereby preventing miR-412-3p-mediated repression of its target protocadherin 7 (PCDH7) in ccRCC cells. Collectively, our results demonstrate that circDVL1 exerts tumor-suppressive function during ccRCC progression through circDVL1/miR-412-3p/PCDH7 axis, and suggest that circDVL1 could be a novel diagnostic and prognositc marker and therapeutic target for ccRCC.     

Efficient assembly of a large fragment of monkeypox virus genome as a qPCR template using dual-selection based transformation-associated recombination

Transformation-associated recombination (TAR) has been widely used to assemble large DNA constructs. One of the significant obstacles hindering assembly efficiency is the presence of error-prone DNA repair pathways in yeast, which results in vector backbone recircularization or illegitimate recombination products. To increase TAR assembly efficiency, we prepared a dual-selective TAR vector, pGFCS, by adding a P_ADH1-URA3 cassette to a previously described yeast-bacteria shuttle vector, pGF, harboring a P_HIS3-HIS3 cassette as a positive selection marker. This new cassette works as a negative selection marker to ensure that yeast harboring a recircularized vector cannot propagate in the presence of 5-fluoroorotic acid. To prevent pGFCS bearing ura3 from recombining with endogenous ura3-52 in the yeast genome, a highly transformable Saccharomyces cerevisiae strain, VL6-48B, was prepared by chromosomal substitution of ura3-52 with a transgene conferring resistance to blasticidin. A 55-kb genomic fragment of monkeypox virus encompassing primary detection targets for quantitative PCR was assembled by TAR using pGFCS in VL6-48B. The pGFCS-mediated TAR assembly showed a zero rate of vector recircularization and an average correct assembly yield of 79% indicating that the dual-selection strategy provides an efficient approach to optimizing TAR assembly.     

Cytoplasmic domain and enzymatic activity of ACE2 are not required for PI4KB dependent endocytosis entry of SARS-CoV-2 into host cells

The recent COVID-19 pandemic poses a global health emergency. Cellular entry of the causative agent SARS-CoV-2 is mediated by its spike protein interacting with cellular receptor-human angiotensin converting enzyme 2 (ACE2). Here, by using lentivirus based pseudotypes bearing spike protein, we demonstrated that entry of SARS-CoV-2 into host cells was dependent on clathrin-mediated endocytosis, and phosphoinositides played essential roles during this process. In addition, we showed that the intracellular domain and the catalytic activity of ACE2 were not required for efficient virus entry. Finally, we showed that the current predominant Delta variant, although with high infectivity and high syncytium formation, also entered cells through clathrin-mediated endocytosis. These results provide new insights into SARS-CoV-2 cellular entry and present proof of principle that targeting viral entry could be an effective way to treat different variant infections.     

野艾蒿挥发油对HeLa癌细胞形态与结构的影响

采用MTT法检测细胞活力,用倒置显微镜、荧光显微镜和扫描电子显微镜观察细胞形态与结构的变化,用激光共聚焦显微镜观察细胞微管的分布,从而研究了野艾蒿挥发油对HeLa人宫颈癌细胞形态与结构的影响。结果表明:(1)野艾蒿挥发油对HeLa癌细胞的增殖有明显的抑制作用,呈剂量和时间依赖性。(2)野艾蒿挥发油处理HeLa癌细胞24h后,100、200μg/mL实验组细胞体积缩小,核染色质凝集、微绒毛消失、细胞表面有泡状突起,微管解聚,呈现典型的凋亡特征;400μg/mL实验组细胞膜破裂、胞浆内含物外泄,呈明显的坏死特征。(3)野艾蒿挥发油具有抑制HeLa癌细胞增殖的作用,低、中浓度的野艾蒿挥发油诱导细胞凋亡,而高浓度的野艾蒿挥发油引起细胞坏死。     

EML4 promotes the loading of NUDC to the spindle for mitotic progression

Echinoderm microtubule-associated protein (EMAP)-like (EML) family proteins are microtubule-associated proteins that have a conserved hydrophobic EMAP-like protein (HELP) domain and multiple WD40 domains. In this study, we examined the role of EML4, which is a member of the EML family, in cell division. Time-lapse microscopy analysis demonstrated that EML4 depletion induced chromosome misalignment during metaphase and delayed anaphase initiation. Further analysis by immunofluorescence showed that EML4 was required for the organization of the mitotic spindle and for the proper attachment of kinetochores to microtubules. We searched for EML4-associating proteins by mass spectrometry analysis and found that the nuclear distribution gene C (NUDC) protein, which is a critical factor for the progression of mitosis, was associated with EML4. This interaction was mediated by the WD40 repeat of EML4 and by the C-terminus of NUDC. In the absence of EML4, NUDC was no longer able to localize to the mitotic spindle, whereas NUDC was dispensable for EML4 localization. Our results show that EML4 is critical for the loading of NUDC onto the mitotic spindle for mitotic progression.     

渤海东部海域秋季底层游泳动物种类组成及群落多样性

根据2014年9月在渤海东部海域(37°40'—38°20'N、120°00'—121°20'E)进行的游泳动物底拖网调查数据,应用相对重要性指数、生物多样性指数、群落结构多元统计分析等方法对该海域游泳动物种类组成及群落多样性特征进行了分析。结果表明:调查海域共捕获游泳动物54种,以头足类枪乌贼(Loliolus spp.)为绝对优势种;游泳动物资源密度为28.82kg/h和5166尾/h。游泳动物种类Margalef丰富度指数(D)的变化范围为1.99—3.67,Shannon-Wiener多样性指数(H')变化范围为0.89—2.28,Pielou均匀度指数(J')变化范围为0.29—0.69。群落结构多元统计分析表明,渤海东部海域游泳动物群落结构以60%的相似性可划分为3个组群:长岛群岛海域组群(A组群)、龙口海域组群(B组群)和蓬莱海域组群(C组群);ANOSIM分析表明,组群之间差异极显著,两两之间差异亦极显著。受增殖放流影响,C组群资源密度和种类多样性均保持较高的水平,而A、B组群人为活动频繁,过度捕捞严重,对海域生态环境和渔业资源整体结构的破坏较大。     

羊血铜锌超氧化物歧化酶(Cu-ZnSOD)的分离纯化

目的:对羊血进行提取SOD。方法:采用有机溶剂去除血红蛋白、热变性、丙酮沉淀、DEAE-32离子交换柱层析的方法,对羊红细胞Cu,Zn-SOD进行分离纯化。利用非变性凝胶电泳NBT活性染色鉴定SOD,SDS-PAGE测定分子量。结果:表明1 000ml羊血中得到SOD干粉1 257mg,总活力为79 453U,比活力为3 871.4U/mg。活性染色结果证明羊血SOD有2条带,表明已达到电泳纯。SDS-PAGE测得SOD亚基分子量分别为16.71kDa、15.97kDa。H2O2对羊血CuZn-SOD活性有抑制作用,通过紫外光谱扫描,羊血SOD在231nm处有最大吸收峰。     

Disruption of Ssp411 causes impaired sperm head formation and male sterility in mice

Background

We previously cloned the Ssp411 gene. We found that the Ssp411 protein is predominantly expressed in elongated spermatids in the rat testis in a stage-dependent manner. Although our findings strongly suggested that Ssp411 might play an important role in mammalian spermatogenesis, this hypothesis has not been studied.

New contributions to the biodiversity of ciliates (Protozoa,Ciliophora) from Antarctica,including a description of <Emphasis Type="Italic">Gastronauta multistriata</Emphasis> nov. spec.

Antarctica is a remote and isolated biotope which makes it an ideal location for studying new and endemic species. Since there is little literature available on the diversity of ciliates in this area, a taxonomic survey of ciliates from melt-water of Collins glacier, King George Island, was carried out from January to March 2006. As a result, the morphology and infraciliature of five ciliates, including one new species, are described using live observations and silver staining: Gastronauta multistriata nov. spec., Neokeronopsis asiatica Foissner et al., 2010, Paraholosticha muscicola Kahl, 1932, Oxytricha sp., and Cyclidium glaucoma Müller, 1786. Gastronauta multistriata nov. spec. is distinguished from its congeners by the following combination of characters: cell size in vivo on average 80 × 40 μm; 5–9 kineties in left ciliary field; 18–23 kineties in right ciliary field, including 10–12 postoral kineties; 7–10 preoral kineties; dorsal brush along anterior dorsal margin, consisting of 5–8 groups of basal bodies. The only minor differences between the current population of N. asiatica and a previously described Antarctic population are the numbers of caudal cirri (6–10 vs. 8–15) and dorsal kineties (11–13 vs. 12–18). Paraholosticha sterkii is synonymised with P. muscicola. The Antarctic population of C. glaucoma corresponds well with a former population from China, the only difference being the number of kinetids in SK _n (11–17 vs. 9–11). This work will contribute to the understanding of ciliate diversity in this little studied area.