青虾生长性状相关的微卫星标记筛选

青虾,学名日本沼虾(Macrobrachium nipponense),又称河虾.属于甲客纲(Crustacea),十足目(Decapoda),长臂虾科(Palaemonidae),沼虾属(Macrobrachium)[1].具有适应性强、分布广、食性杂、生长快、养殖经济效益高等特点,是我国淡水虾类的一个重要养殖品种[2].也是主要淡水名特优养殖品种之一.目前,我国养殖青虾出现了个体小型化、性早熟以及生长速率慢等种质退化现象,极大地影响了日本沼虾养殖业的效益,迫切需要对其进行遗传改良[3,4].     

利用烟草瞬时表达系统检测高等植物基因的剪接加工

解析基因的剪接加工机制是了解植物形态建成、生长发育和逆境胁迫应答的重要环节．与动物相比,植物中相应的研究进展较为缓慢．利用农杆菌介导的烟草瞬时表达系统,分别对单子叶植物水稻BADH2和双子叶植物拟南芥GR7基因片段在烟草叶片中的转录后剪接加工进行分析．结果表明,一些重要剪接调控元件在植物中保守存在,而烟草瞬时表达系统可以作为研究高等植物剪接调控的重要工具,快捷灵敏地检测基因的剪接加工方式．     

Increases in cerebrovascular impedance in older adults

This study explored a novel method for measuring cerebrovascular impedance to quantify the relationship between pulsatile changes in cerebral blood flow (CBF) and arterial pressure. Arterial pressure in the internal or common carotid artery (applanation tonometry), CBF velocity in the middle cerebral artery (transcranial Doppler), and end-tidal CO(2) (capnography) were measured in six young (28 ± 4 yr) and nine elderly subjects (70 ± 6 yr). Transfer function method was used to estimate cerebrovascular impedance. Under supine resting conditions, CBF velocity was reduced in the elderly despite the fact that they had higher arterial pressure than young subjects. As expected, cerebrovascular resistance index was increased in the elderly. In both young and elderly subjects, impedance modulus was reduced gradually in the frequency range of 0.78-8 Hz. Phase was negative in the range of 0.78-4.3 Hz and fluctuated at high frequencies. Compared with the young, impedance modulus increased by 38% in the elderly in the range of 0.78-2 Hz and by 39% in the range of 2-4 Hz (P < 0.05). Moreover, increases in impedance were correlated with reductions in CBF velocity. Collectively, these findings demonstrate the feasibility of assessing cerebrovascular impedance using the noninvasive method developed in this study. The estimated impedance modulus and phase are similar to those observed in the systemic circulation and other vascular beds. Moreover, increases in impedance in the elderly suggest that arterial stiffening, besides changes in cerebrovascular resistance, contributes to reduction in CBF with age.     

Genetic variations in plasma circulating DNA of HBV-related hepatocellular carcinoma patients predict recurrence after liver transplantation

Background

Recurrence prediction of hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC) patients undergoing liver transplantation (LT) present a great challenge because of a lack of biomarkers. Genetic variations play an important role in tumor development and metastasis.

Methods

Oligonucleotide microarrays were used to evaluate the genetic characteristics of tumor DNA in 30 HBV-related HCC patients who were underwent LT. Recurrence-related single-nucleotide polymorphism were selected, and their prognostic value was assessed and validated in two independent cohorts of HCC patients (N = 102 and N = 77), using pretransplant plasma circulating DNA. Prognostic significance was assessed by Kaplan-Meier survival estimates and log-rank tests. Multivariate analyses were performed to evaluate prognosis-related factors.

Results

rs894151 and rs12438080 were significantly associated with recurrence (P = .003 and P = .004, respectively). Multivariate analyses demonstrated that the co-index of the 2 SNPs was an independent prognostic factor for recurrence (P = .040). Similar results were obtained in the third cohort (N = 77). Furthermore, for HCC patients (all the 3 cohorts) exceeding Milan criteria, the co-index was a prognostic factor for recurrence and survival (P<.001 and P = .002, respectively).

Conclusions

Our study demonstrated first that genetic variations of rs894151 and rs12438080 in pretransplant plasma circulating DNA are promising prognostic markers for tumor recurrence in HCC patients undergoing LT and identify a subgroup of patients who, despite having HCC exceeding Milan criteria, have a low risk of post-transplant recurrence.     

亚洲古近纪早期的年代学和哺乳动物群更替(英文)

对广东南雄盆地、江西池江盆地、安徽潜山盆地、湖南衡阳盆地和内蒙古二连盆地古新世和早始新世沉积物进行的生物地层学、化学地层学和磁性地层学研究,第一次给亚洲早古近纪地层框架提供了严格的年代限定。亚洲古新世和早始新世地层划分为4个生物年代单位(陆生哺乳动物期),从老到新为上湖期、浓山期、格沙头期和伯姆巴期4个亚洲陆生哺乳动物期。南雄盆地古地磁数据显示上湖期的底界约位于C29r的上2/3位置。南雄盆地的古地磁资料及池江盆地最新的古地磁和同位素结果显示上湖-浓山期界线在C27n的上部和C26r的下部之间,接近C27n-C26r转换界面。这一记录支持将上湖-浓山期的界线与北美陆生哺乳动物期中的Torrejonian-Tiffanian期以及海相记录中丹尼-塞兰特期(Danian-Selandi-an)的界线相对比。二连盆地古地磁和同位素研究表明典型的格沙头动物群延续至C24r,并且可能向下延伸至C25r和C26n。南雄盆地的记录指示浓山组和古城村组可能相当于C26r的上部。这些结果表明亚洲哺乳动物分期中的浓山-格沙头期的界线可能在北美哺乳动物分期的Tiffanian期中。衡阳盆地的古地磁和同位素证据指示,以短暂的碳同位素漂移为标志的古新世-始新世界线出现在栗木坪组和岭茶组之间,位于C24r中,与海相记录中的坦尼特-伊普里斯期(Thanetian-Ypresian)界线一致。伯姆巴期岭茶动物群的时代似乎与北美Wa-satchian-0带的动物群可以比较。上湖期和浓山期动物群主要由亚洲特有的动物组成。发生在上湖期和浓山期界线处的动物群更替,与北美Torrejonian-Tiffanian期界线以及丹尼-塞兰特期界线一致。在相隔很远的生态系统上的这种巧合,可能反映出各自独立的对气候变化的生态和/或进化上的响应。格沙头动物群的地方性色彩更弱,有与北美Clarkforkian期动物群共有的属一级分类单元。伯姆巴动物群是全球性的。现生哺乳动物各目在格沙头-伯姆巴期界线处突然出现,与北美和欧洲的记录一致,显示动物群的更替与古新世-始新世界线处短暂出现的全球变暖事件(又称古新世-始新世高温事件)有关。     

Codon modification for the DNA sequence of a single-chain Fv antibody against clenbuterol and expression in Pichia pastoris

The expression efficiency was improved for the recombinant single-chain variable fragment (scFv) against clenbuterol (CBL) obtained from mouse and expressed in the methylotrophic yeast Pichia pastoris GS115, by redesigning and synthesizing the DNA sequence encoding for CBL-scFv based on the codon bias of P. pastoris. The codons enco4ding 124 amino acids were optimized, in which a total of 156 nucleotides were changed, and the G+C ratio was simultaneously decreased from 53 to 47.2 %. Under the optimized expression conditions, the yield of the recombinant CBL-scFv (41 kDa) antibodies was 0.223 g L^–1 in shake culture. Compared to the non-optimized control, the expression level of the optimized recombinant CBL-scFv based on preferred codons in P. pastoris demonstrated a 2.35-fold higher yield. Furthermore, the recombinant CBL-scFv was purified by Ni-NTA column chromatography, and the purity was 95 %. The purified CBL-scFv showed good CBL recognition by a competitive indirect enzyme-linked immunoassay. The average concentration required for 50 % inhibition of binding and the limit of detection for the assay were 5.82 and 0.77 ng mL^–1, respectively.     

PPP2R5C Couples Hepatic Glucose and Lipid Homeostasis

In mammals, the liver plays a central role in maintaining carbohydrate and lipid homeostasis by acting both as a major source and a major sink of glucose and lipids. In particular, when dietary carbohydrates are in excess, the liver converts them to lipids via de novo lipogenesis. The molecular checkpoints regulating the balance between carbohydrate and lipid homeostasis, however, are not fully understood. Here we identify PPP2R5C, a regulatory subunit of PP2A, as a novel modulator of liver metabolism in postprandial physiology. Inactivation of PPP2R5C in isolated hepatocytes leads to increased glucose uptake and increased de novo lipogenesis. These phenotypes are reiterated in vivo, where hepatocyte specific PPP2R5C knockdown yields mice with improved systemic glucose tolerance and insulin sensitivity, but elevated circulating triglyceride levels. We show that modulation of PPP2R5C levels leads to alterations in AMPK and SREBP-1 activity. We find that hepatic levels of PPP2R5C are elevated in human diabetic patients, and correlate with obesity and insulin resistance in these subjects. In sum, our data suggest that hepatic PPP2R5C represents an important factor in the functional wiring of energy metabolism and the maintenance of a metabolically healthy state.     

Asymmetric Engagement of Amygdala and Its Gamma Connectivity in Early Emotional Face Processing

The amygdala has been regarded as a key substrate for emotion processing. However, the engagement of the left and right amygdala during the early perceptual processing of different emotional faces remains unclear. We investigated the temporal profiles of oscillatory gamma activity in the amygdala and effective connectivity of the amygdala with the thalamus and cortical areas during implicit emotion-perceptual tasks using event-related magnetoencephalography (MEG). We found that within 100 ms after stimulus onset the right amygdala habituated to emotional faces rapidly (with duration around 20–30 ms), whereas activity in the left amygdala (with duration around 50–60 ms) sustained longer than that in the right. Our data suggest that the right amygdala could be linked to autonomic arousal generated by facial emotions and the left amygdala might be involved in decoding or evaluating expressive faces in the early perceptual emotion processing. The results of effective connectivity provide evidence that only negative emotional processing engages both cortical and subcortical pathways connected to the right amygdala, representing its evolutional significance (survival). These findings demonstrate the asymmetric engagement of bilateral amygdala in emotional face processing as well as the capability of MEG for assessing thalamo-cortico-limbic circuitry.     

Origins of weedy rice revealed by polymorphisms of chloroplast DNA sequences and nuclear microsatellites

Conspecific weeds that permanently infest worldwide agroecosystems are evolved from their crop species. These weeds cause substantial problems for crop production by competing for resources in agricultural fields. Weedy rice represents such a conspecific weed infesting rice ecosystems, and causing tremendous rice yield losses owing to its strong competitiveness and abundant genetic diversity, likely resulted from its complex origins. Here, we report the use of chloroplast DNA (cpDNA) fingerprints to determine whether weedy rice is evolved from its wild (exo‐feral) or cultivated (endo‐feral) rice progenitor as the maternal donor in recent hybridization events. In addition, we also applied nuclear simple sequence repeat (SSR) markers to confirm the exo‐feral or endo‐feral origins of weedy rice accessions determined by the cpDNA fingerprints. We found that the studied weedy rice accessions evolved either from their wild or cultivated rice progenitor, as the maternal donor, based on the cpDNA network and structure analyses. Combined analyses of cpDNA and nuclear SSR markers indicated that a much greater proportion of weedy rice accessions had the endo‐feral origin. In addition, results from the genetic structure of nuclear SSR markers indicated that weedy rice accessions from the endo‐feral pathway are distinctly associated with either indica or japonica rice cultivars, suggesting their complex origins through crop–weed introgression. The complex pathways of origin and evolution could greatly promote genetic diversity of weedy rice. Therefore, innovative methods should be developed for effective weedy rice control.     

Effects of simulated drought stress on carotenoid contents and expression of related genes in carrot taproots

Carotenoids are liposoluble pigments found in plant chromoplasts that are responsible for the yellow, orange, and red colors of carrot taproots. Drought is one of the main stress factors affecting carrot growth. Carotenoids play important roles in drought resistance in higher plants. In the present work, the carotenoid contents in three different-colored carrot cultivars, ‘Kurodagosun’ (orange), ‘Benhongjinshi’ (red), and ‘Qitouhuang’ (yellow), were determined by ultra-high-performance liquid chromatography (UPLC) after 15% polyethylene glycol (PEG) 6000 treatment. Real-time fluorescence quantitative PCR (RT-qPCR) was then used to determine the expression levels of carotenoid synthesis- and degradation-related genes. Increases in β-carotene content in ‘Qitouhuang’ taproots under drought stress were found to be related to the expression levels of DcPSY2 and DcLCYB. Increases in lutein and decreases in α-carotene content in ‘Qitouhuang’ and ‘Kurodagosun’ under PEG treatment may be related to the expression levels of DcCYP97A3, DcCHXE, and DcCHXB1. The expression levels of DcNCED1 and DcNCED2 in the three cultivars significantly increased, thus suggesting that NCED genes could respond to drought stress. Analysis of the growth status and carotenoid contents of carrots under PEG treatment indicated that the orange cultivar ‘Kurodagosun’ has better adaptability to drought stress than the other cultivars and that β-carotene and lutein may be involved in the stress resistance process of carrot.