多能性转录因子OCT4和SOX2在体外发育的小鼠2-细胞胚胎的表达与定位

为探讨多能性转录因子OCT4和SOX2在昆明小鼠(Mus musculus)2-细胞胚胎发育过程中与2-细胞胚胎阻滞发生的相关性,本研究应用实时荧光定量PCR技术检测了小鼠卵母细胞及在M16培养液中培养的不同发育阶段体外受精胚Oct4和Sox2基因的表达,并利用实时荧光定量PCR和免疫荧光技术比较了2-细胞胚、2-细胞阻滞胚和4-细胞胚的OCT4和SOX2的表达与定位。采用ANOVA对实验所得的数据进行分析,P0.05被认为是具有显著性差异。研究结果显示,2-细胞胚只有24.8%发育成4-细胞胚,75.2%的2-细胞胚发生了阻滞。Sox2和Oct4的m RNA在MⅡ期卵母细胞、原核胚、2-细胞胚、4-细胞胚、桑椹胚和囊胚中都有表达。Oct4 m RNA的表达水平在4-细胞胚显著高于2-细胞胚和2-细胞阻滞胚(P0.05),Sox2 m RNA的表达水平在2-细胞胚显著高于2-细胞阻滞胚和4-细胞胚(P0.05),而后两者之间没有差异(P0.05)。OCT4蛋白在2-细胞胚和4-细胞胚中与核共定位,但在2-细胞阻滞胚中弥散存在于胞质中。SOX2蛋白在以上3类胚胎中始终定位于细胞核。上述结果提示,转录因子OCT4和SOX2的表达和定位与小鼠2-细胞胚胎发育阻滞相关,母源性SOX2表达的维持对胚胎合子基因组激活(ZGA)的发生具有重要作用,母源性OCT4的异常定位可能影响了合子基因组激活相关基因的激活,而合子中Oct4的表达影响合子基因组激活后胚胎的发育。     

过表达微小RNA miR-125b抑制肝癌细胞上皮-间质转换及促进细胞凋亡

微小RNA-125b（miR-125b）在许多恶性肿瘤的增殖、分化和凋亡等过程中具有很重要的作用,但miR-125b是否涉及肝癌的上皮 间质转换过程（EMT）还有待进一步研究。本研究通过构建过表达miR-125b的肝癌稳转细胞株,初步检测miR-125b对于肝癌的EMT过程和相关的TGF-β信号通路的影响,以及对于肝癌细胞凋亡的影响。以慢病毒载体pHRS-1cla EGFP 构建过表达miR-125b的载体质粒(pHRS-1cla-miR125b-CMV-EGFP),并对上述载体进行NheⅠ、XbaⅠ双酶切和测序鉴定,鉴定正确后,在293T细胞中进行慢病毒包装,浓缩病毒后,对MHCC97-H进行慢病毒感染并采用流式分选GFP阳性的细胞。实时定量PCR检测表明肝癌细胞稳转株MHCC97-H-PHRS-miR-125b-EGFP的miR-125b表达量是空载体转染组的6倍。Western印迹检测发现,与空载体对照组相比,MHCC97-H-PHRS-miR-125b-EGFP细胞中间质细胞标志α-SMA表达显著下调,上皮细胞标志E-cadherin表达显著上调,同样的,用Western印迹检测也发现MHCC97-H-PHRS-miR-125b-EGFP细胞中TGF-β信号通路关键下游分子Smad2和Smad4的表达显著下调,细胞凋亡检测结果表明,与对照组相比,过表达miR-125b的稳转株凋亡率增加到19.66%,加入TGF-β1后,过表达miR-125b的稳转株凋亡率进一步增加到74.7%。同样的,在体内治疗实验中,我们采用商品化的体内核酸转染试剂,在皮下肿瘤组织中过表达miR-125b mimics,结果表明miR-125b的过表达与肿瘤组织的凋亡成正相关性（r=0.83463,P < 0.01）,且免疫组化结果也表明,miR-125b过表达后,E-cadherin表达显著上调,α-SMA及Smad2和Smad4的表达显著下调。上述结果表明,我们成功构建了过表达miR-125b的肝癌细胞稳转株,并成功建立了肿瘤组织中过表达miR-125b mimics的动物模型,在体内外均观察到过表达miR-125b后对肝癌细胞EMT过程的抑制作用和对细胞凋亡的促进作用。相关研究结果加深了我们对miR-125b在肝癌中抑制肝癌发展作用机制的理解,及其作为潜在的治疗肝癌的新靶点的重要性。     

A Preliminary Study on Alien Invasive Plants in Jiangsu,China

From 1985 to 2013, there were only 9 literatures reports on alien invasive plants in Jiangsu, less than 025% of total 3665 similar reports during the same period in China, and it was much disproportionate with the development level of social, economic, cultural and other aspects of the province. Through field investigation, literature review and specimen inspection, the preliminary statistics, analysis and discussion of 47 families 128 genera and 209 species of alien invasive plants in Jiangsu have been made, including their composition, origin, and the life form. All alien invasive plants were ranked into 5 grades according to the damage to ecosystem and economy: twenty two malignant invasive species; forty one serious invasive species; thirty two local invasive species; fifty one general invasive species; a left 63 species with unknown damage wait for further observation. The characteristics of alien invasive plants in Jiangsu have been discussed, and the proposals of suggestion have been made.     

Evolutionary history of a desert perennial Arnebia szechenyi (Boraginaceae): Intraspecific divergence,regional expansion and asymmetric gene flow

The complex interactions of historical, geological and climatic events on plant evolution have been an important research focus for many years. However, the role of desert formation and expansion in shaping the genetic structures and demographic histories of plants occurring in arid areas has not been well explored. In the present study, we investigated the phylogeography of Arnebia szechenyi, a desert herb showing a near-circular distribution surrounding the Tengger Desert in Northwest China. We measured genetic diversity of populations using three maternally inherited chloroplast DNA (cpDNA) fragments and seven bi-paternally inherited nuclear DNA (nDNA) loci that were sequenced from individuals collected from 16 natural populations across its range and modelled current and historical potential habitats of the species. Our data indicated a considerably high level of genetic variation within A. szechenyi and noteworthy asymmetry in historical migration from the east to the west. Moreover, two nuclear genetic groups of populations were revealed, corresponding to the two geographic regions separated by the Tengger Desert. However, analysis of cpDNA data did not show significant geographic structure. The most plausible explanation for the discrepancy between our findings based on cpDNA and nDNA data is that A. szechenyi populations experienced long periods of geographic isolation followed by range expansion, which would have promoted generalized recombination of the nuclear genome. Our findings further highlight the important role that the Tengger Desert, together with the Helan Mountains, has played in the evolution of desert plants and the preservation of biodiversity in arid Northwest China.     

新疆南部棉区杏树对棉田主要害虫和自然天敌的影响

2008—2011年在新疆莎车县调查研究了杏树对棉田主要害虫及自然天敌种群发生的影响。研究表明杏棉间作田主要害虫及天敌种类与棉单作田一致。杏树间作种植模式对棉田牧草盲蝽Lygus pratensis、棉蓟马Thripstabaci和棉长管蚜Acyrthosiphon gossyfii的种群发生有一定影响,不利于其在田间发生;对棉蚜Aphis gossyfii、棉叶螨Tetrangchus urticae影响不大。棉田瓢虫和食虫蝽天敌受间作和棉蚜发生量的双重影响。间作有利于蜘蛛数量的增加。     

浙江省外来入侵植物研究

基于国内外文献资料、标本信息和野外调查,初步整理了浙江省外来入侵植物的种类组成、生活型、原产地、危害程度和入侵途径。结果表明,浙江省现有外来人侵植物46科106属166种,种数最多的是菊科、豆科和禾本科。根据原产地分析,来源于南美洲的物种最多,约占总数的32．16％,其次为北美洲,约占总数的25．63％,欧洲排第三位,约占总数的19．59％;已产生严重危害的有23种,中等危害的60种,危害较轻的83种。还有一些物种虽然没有在表里列出,但它们具有潜在的危害,必须密切关注。同时,与之前的研究相比,我们去除了一些已报道为人侵的物种,又增加了一些新的入侵种。在此基础上,对浙江省外来入侵植物的生活型、来源及危害进行了分析,并与以往的研究进行了比较,提出了针对性的管理建议。     

科尔沁沙地沙丘迎风坡面干沙层的空间分布

对科尔沁沙地西部地区流动沙丘和固定沙丘的迎风坡表层干沙层厚度进行观测,分析了沙丘坡面干沙层的空间分布特点.结果表明:研究区沙丘迎风坡表层干沙层厚度主要在5 ～15 cm,92.0％的流动沙丘区域和98.6％的固定沙丘区域的干沙层厚度分布在此范围内;固沙植物影响沙丘表层干沙层的厚度变化和空间分布.流动沙丘表层干沙层厚度表现出明显的空间差异性:迎风坡上部西侧区域干沙层厚度较大,下部东侧区域相对较薄,其平均厚度为(9.58±3.95) cm,干沙层厚度在0～ 40 cm范围内变化,变异系数为41％,干沙层空间分布的变异函数表现为球状模型,具有中等的空间相关性;固定沙丘的干沙层厚度表现为明显的均一性,空间差异性不明显,其平均厚度为(10.91±1.70) cm,干沙层厚度变化范围在0～20 cm,变异系数仅16％.     

Cytotoxic steroidal saponins from Trillium kamtschaticum

Eight new steroidal saponins, trillikamtosides K–R (1–8), along with three known analogues, were isolated from the whole plants of Trillium kamtschaticum. Their structures were unambiguously established by interpretation of spectroscopic data (MS and NMR) and chemical methods. Compound 1 had a rare aglycone featuring a skeleton of 16-oxaandrost-5-en-3-ol-17-one, which was reported for the first time. The isolated saponins were tested for cytotoxicities against HCT116 cells, and trillikamtoside R (8) was found to show the most cytotoxic effect with an IC₅₀ value of 4.92 μM.     

Nitrogen deposition promotes phosphorus uptake of plants in a semi-arid temperate grassland

Plant and Soil - Nitrogen (N) deposition greatly influences ecosystem processes through the alteration of plant nutrition; however, there is limited understanding about the effects of phosphorus...     

Clinical characteristics and risk factors for severe scrub typhus in pediatric and elderly patients

BackgroundScrub typhus (ST) is a life-threatening infectious disease if appropriate treatment is unavailable. Large discrepancy of clinical severity of ST patients was reported among age groups, and the underlying risk factors for severe disease are unclear.MethodsClinical and epidemiological data of ST patients were collected in 55 surveillance hospitals located in Guangzhou City, China, from 2012 to 2018. Severe prognosis and related factors were determined and compared between pediatric and elderly patients.ResultsA total of 2,074 ST patients including 209 pediatric patients and 1,865 elderly patients were included, with a comparable disease severity rate of 11.0% (95% CI 7.1%–16.1%) and 10.3% (95% CI 9.0%–11.8%). Different frequencies of clinical characteristics including lymphadenopathy, skin rash, enlarged tonsils, etc. were observed between pediatric and elderly patients. Presence of peripheral edema and decreased hemoglobin were the most important predictors of severe illness in pediatric patients with adjusted ORs by 38.99 (9.96–152.67, p<0.001) and 13.22 (1.54–113.50, p = 0.019), respectively, while presence of dyspnea and increased total bilirubin were the potential determinants of severe disease in elderly patients with adjusted ORs by 11.69 (7.33–18.64, p<0.001) and 3.17 (1.97–5.11, p<0.001), respectively. Compared with pediatric patients, elderly patients were more likely to receive doxycycline (64.8% v.s 9.9%, p<0.001), while less likely to receive azithromycin therapy (5.0% v.s 41.1%, p<0.001).ConclusionThe disease severity rate is comparable between pediatric and elderly ST patients, while different clinical features and laboratory indicators were associated with development of severe complications for pediatric and elderly patients, which is helpful for diagnosis and progress assessment of disease for ST patients.