circPVT1通过miR-24-3p/let-7a-5p/FSCN1轴促进鼻咽癌细胞侵袭迁移

目的 鼻咽癌是一种来源于鼻咽上皮的恶性肿瘤,其临床特征之一是易发生淋巴转移,但是目前鼻咽癌转移的分子机制尚未阐明。circPVT1是由PVT1基因2号外显子反向拼接形成的环状RNA (circRNA),在多种肿瘤中表达上调,本文探讨了circPVT1在鼻咽癌侵袭迁移中的作用和分子机制。方法 通过RT-qPCR检测circPVT1及其下游miRNA和FSCN1在鼻咽癌细胞的表达情况,Transwell和划痕愈合实验检测circPVT1对鼻咽癌细胞侵袭迁移的影响,RNA pull-down实验检测circPVT1结合的miRNA,双荧光素酶报告实验检测miR-24-3p和let-7a-5p靶向抑制FSCN1 mRNA表达。结果 在鼻咽癌细胞中过表达circPVT1可以促进鼻咽癌细胞侵袭迁移,而敲低circPVT1则可以抑制鼻咽癌细胞的侵袭迁移。进一步研究发现,circPVT1可以通过竞争性吸附miR-24-3p和let-7a-5p,上调FSCN1的表达,从而促进鼻咽癌细胞的侵袭迁移。结论 circPVT1通过miR-24-3p/let-7a-5p/FSCN1轴促进鼻咽癌细胞侵袭迁移,证实c...     

Prp5 bridges U1 and U2 snRNPs and enables stable U2 snRNP association with intron RNA

Communication between U1 and U2 snRNPs is critical during pre-spliceosome assembly; yet, direct connections have not been observed. To investigate this assembly step, we focused on Prp5, an RNA-dependent ATPase of the DExD/H family. We identified homologs of Saccharomyces cerevisiae Prp5 in humans (hPrp5) and Schizosaccharomyces pombe (SpPrp5), and investigated their interactions and function. Depletion and reconstitution of SpPrp5 from extracts demonstrate that ATP binding and hydrolysis by Prp5 are required for pre-spliceosome complex A formation. hPrp5 and SpPrp5 are each physically associated with both U1 and U2 snRNPs; Prp5 contains distinct U1- and U2-interacting domains that are required for pre-spliceosome assembly; and, we observe a Prp5-associated U1/U2 complex in S. pombe. Together, these data are consistent with Prp5 being a bridge between U1 and U2 snRNPs at the time of pre-spliceosome formation.     

重铬酸钾对蚕豆根尖细胞致畸效应的研究

以蚕豆根尖为材料,研究重铬酸钾对蚕豆根尖细胞的致畸效应。采用蚕豆根尖细胞的微核试验和染色体畸变试验方法,以不同浓度的重铬酸钾为诱变剂,测定蚕豆根尖细胞的微核率和染色体畸变率。结果表明：重铬酸钾能诱发较高频率的微核率,即在一定浓度范围内,其微核率随重铬酸钾处理浓度的升高而增加,但高于一定浓度后反而呈下降趋势;不同浓度的重铬酸钾均使蚕豆根尖细胞有丝分裂指数增大;重铬酸钾还能诱导蚕豆根尖细胞产生较高频率的染色体畸变,且产生多种类型的染色体畸变。结论是重铬酸钾对蚕豆根尖细胞具有明显的致畸效应。Abstract：We studied the aberrant effects of different concentrations of potassium dichromate on Vicia Faba root tip cells. The micronucleus and chromosome aberration assay was conducted to determine the micronucleus rate and chromosome aberration rate of Vicia faba root tip cells induced by potassium dichromate. The result indicated that potassium dichromate could increase the micronucleus rate of Vicia faba root tip cells. Within certain range of concentration the rate of micronucleus was found to be increased with the increase of potassium dichromate concentration,but beyond this range the rate of micronucleus decreased with further increase of potassium dichromate concentration. The potassium dichromate at different concentrations could increase the cell mitosis index. Besides,it also caused various types of chromosome aberration,and the rates of chromosome aberration were always higher than that of the control group. The conclusion of this study was that potassium dichromate has obvious teratogenic effect on vicia faba root tip cells.     

Effect of low temperature on globin expression,respiratory metabolic enzyme activities,and gill structure of <Emphasis Type="Italic">Litopenaeus vannamei</Emphasis>

Low temperature frequently influences growth, development, and even survival of aquatic animals. In the present study, physiological and molecular responses to low temperature in Litopenaeus vannamei were investigated. The cDNA sequences of two oxygen-carrying proteins, cytoglobin (Cygb) and neuroglobin (Ngb), were isolated. Protein structure analysis revealed that both proteins share a globin superfamily domain. Real-time PCR analysis indicated that Cygb and Ngb mRNA levels gradually increased during decrease in temperatures from 25 to 15°C and then decreased at 10°C in muscle, brain, stomach, and heart, except for a continuing increase in gills, whereas they showed a different expression trend in the hepatopancreas. Hemocyanin concentration gradually reduced as the temperature decreased. Moreover, the activities of respiratory metabolic enzymes including lactate dehydrogenase (LDH) and succinate dehydrogenase (SDH) were measured, and it was found that LDH activity gradually increased while SDH activity decreased after low-temperature treatment. Finally, damage to gill structure at low temperature was also observed, and this intensified with further decrease in temperature. Taken together, these results show that low temperature has an adverse influence in L. vannamei, which contributes to systematic understanding of the adaptation mechanisms of shrimp at low temperature.     

凡纳滨对虾MT基因序列及其在卵巢发育和低温胁迫中的表达分析

在低温处理仔虾全长cDNA文库的筛选测序中, 获得凡纳滨对虾(Litopenaeus vannmei)金属硫蛋白基因全长cDNA序列, 该序列含有425个碱基, 包含177 bp开放阅读框, 上游98 bp的非编码区及下游150 bp 的非编码区, 编码58个氨基酸, 其中半胱氨酸含量丰富, 富含金属硫蛋白典型的Cys-X(1-3)-Cys 结构。多序列比对表明, 凡纳滨对虾MT蛋白序列与美洲螯龙虾(Homarus americanus)MT蛋白序列具最高同源性72.4%。Real-time PCR结果表明, 凡纳滨对虾MT基因在卵巢组织中呈优势表达, 在不同发育期的卵巢中的表达量都很高, 在低温处理凡纳滨对虾肝胰腺组织中上调表达。实验所得结果为研究凡纳滨对虾金属硫蛋白基因在生殖发育和低温应激中的功能提供了参考。       

Mapping and genotypic analysis of the NK-lysin gene in chicken

Background

Antimicrobial peptides (AMP) are important elements of the first line of defence against pathogens in animals. NK-lysin is a cationic AMP that plays a critical role in innate immunity. The chicken NK-lysin gene has been cloned and its antimicrobial and anticancer activity has been described but its location in the chicken genome remains unknown. Here, we mapped the NK-lysin gene and examined the distribution of a functionally significant single nucleotide polymorphism (SNP) among different chicken inbred lines and heritage breeds.

Results

A 6000 rad radiation hybrid panel (ChickRH6) was used to map the NK-lysin gene to the distal end of chromosome 22. Two additional genes, the adipocyte enhancer-binding protein 1-like gene (AEBP1) and the DNA polymerase delta subunit 2-like (POLD2) gene, are located in the same {"type":"entrez-nucleotide","attrs":{"text":"NW_003779909","term_id":"358468969","term_text":"NW_003779909"}}NW_003779909 contig as NK-lysin, and were thus indirectly mapped to chromosome 22 as well. Previously, we reported a functionally significant SNP at position 271 of the NK-lysin coding sequence in two different chicken breeds. Here, we examined this SNP and found that the A allele appears to be more common than the G allele in these heritage breeds and inbred lines.

Conclusions

The chicken NK-lysin gene mapped to the distal end of chromosome 22. Two additional genes, AEBP1 and POLD2, were indirectly mapped to chromosome 22 also. SNP analyses revealed that the A allele, which encodes a peptide with a higher antimicrobial activity, is more common than the G allele in our tested inbred lines and heritage breeds.     

华南重要人工林及地带性森林凋落物酸碱缓冲能力

植物残体可以修正土壤酸度,但其作用机理还有很大争议,本研究评估森林凋落物作为酸碱缓冲体系的能力,它可能是调节土壤酸度的重要机理之一.凋落物材料取自华南重要的人工林类型及一个地带性顶极森林群落,将材料用不同pH酸溶液提取,并用酸碱进行滴定.结果表明,凋落物本身是一个极强的酸碱缓冲体系,它使浸提液酸度保持不变,并对添加的酸碱起显著的缓冲作用.首次发现两个豆科树种凋落物马占相思(Acacia mangiumWilld)与大叶相思(Acacia auriculaiformis A.Cunn)具有很高的pH值(约pH6.0),约高于土壤酸度2个pH单位,每年凋落于地表的枯落物层通过缓冲机理可以提高雨水0.1至0.4pH单位.其它人工林凋落物酸度约为pH4.0,与它们生长的土壤酸度相似.溶液中无机离子组成上的差异不能完全解释凋落物pH格局,但高钠低硝态氮可能是两个豆科树种高pH的一个重要原因.地带性顶极群落12个树种加权酸度为pH3.90,明显低于土壤酸度(pH4.19),因而,凋落物是林下土壤进一步变酸的驱动因素;发现一个种凋落物(山竹子(Garcinia oblongifolia))酸度极低(pH 3.19)并且缓冲力极强,该种植物生长的土壤将难于通过施用石灰等措施对酸度进行调节,它影响下的土壤环境将影响其它植物的入侵.观光木(Tsoongiodendron odorum Chun)凋落物酸度碱向最远离土壤酸度,这种凋落物酸度与环境酸度间的不一致可能对植物形成某种长期生态压力,并成为该种致濒危的一个因素.因而,植物残体是一个极强的酸碱缓冲体系,它以化学缓冲机理可以影响环境酸度,但可能只有少数种通过这一机理使土壤致酸或致碱,因为多数植物残体与土壤酸度基本相似.     

拟南芥细胞壁关联蛋白激酶融合表达及多克隆抗体的制备

为了研究拟南芥中细胞壁关联蛋白激酶1(WAK1)基因功能,通过RT-PCR得到WAK1 cDNA片段,将其亚克隆至pET28a表达载体中进行表达,获得融合蛋白.用纯化的融合蛋白pET28aWAK1免疫家兔,得到抗WAK1抗体.Western blot结果显示该抗体能特异识别在原核表达系统内表达的抗原,抗WAK1的抗体对拟南芥中的WAK1蛋白具有高度的特异性.