不同生境草鱼肠道微生物组成和群落特征分析

[目的]分析不同生境来源的草鱼前肠、中肠和后肠的微生物组成和群落特征.[方法]利用16S rRNA高通量测序技术比较河流、湖泊、高密度池塘养殖与水库低密度养殖4种不同生境来源的草鱼其前、中、后肠的微生物组成和群落特征.[结果]Venn图、稀释性曲线和Alpha指数分析结果显示,前肠微生物群落多样性以养殖生境草鱼更高,而...     

Diet drives convergent evolution of gut microbiomes in bamboo-eating species

Gut microbiota plays a critical role in host physiology and health. The coevolution between the host and its gut microbes facilitates animal adaptation to its specific ecological niche. Multiple factors such as host diet and phylogeny modulate the structure and function of gut microbiota. However, the relative contribution of each factor in shaping the structure of gut microbiota remains unclear. The giant(Ailuropoda melanoleuca) and red(Ailurus styani) pandas belong to different families of order Carnivora. They have evolved as obligate bamboo-feeders and can be used as a model system for studying the gut microbiome convergent evolution. Here, we compare the structure and function of gut microbiota of the two pandas with their carnivorous relatives using 16S rRNA and metagenome sequencing. We found that both panda species share more similarities in their gut microbiota structure with each other than each species shares with its carnivorous relatives. This indicates that the specialized herbivorous diet rather than host phylogeny is the dominant driver of gut microbiome convergence within Arctoidea.Metagenomic analysis revealed that the symbiotic gut microbiota of both pandas possesses a high level of starch and sucrose metabolism and vitamin B12 biosynthesis. These findings suggest a diet-driven convergence of gut microbiomes and provide new insight into host-microbiota coevolution of these endangered species.     

Enhanced Orai1-mediated store-operated Ca2+ channel/calpain signaling contributes to high glucose-induced podocyte injury

Podocyte injury induced by hyperglycemia is the main cause of kidney dysfunction in diabetic nephropathy. However, the underlying mechanism is unclear. Store-operated Ca²⁺ entry (SOCE) regulates a diversity of cellular processes in a variety of cell types. Calpain, a Ca²⁺-dependent cysteine protease, was recently shown to be involved in podocyte injury. In the present study, we sought to determine whether increased SOCE contributed to high glucose (HG)–induced podocyte injury through activation of the calpain pathway. In cultured human podocytes, whole-cell patch clamp indicated the presence of functional store-operated Ca²⁺ channels, which are composed of Orai1 proteins and mediate SOCE. Western blots showed that HG treatment increased the protein abundance of Orai1 in a dose-dependent manner. Consistently, calcium imaging experiments revealed that SOCE was significantly enhanced in podocytes following HG treatment. Furthermore, HG treatment caused overt podocyte F-actin disorganization as well as a significant decrease in nephrin protein abundance, both of which are indications of podocyte injury. These podocyte injury responses were significantly blunted by both pharmacological inhibition of Orai1 using the small molecule inhibitor BTP2 or by genetic deletion of Orai1 using CRISPR-Cas9 lentivirus. Moreover, activation of SOCE by thapsigargin, an inhibitor of Ca²⁺ pump on the endoplasmic/sarcoplasmic reticulum membrane, significantly increased the activity of calpain, which was inhibited by BTP2. Finally, the calpain-1/calpain-2 inhibitor calpeptin significantly blunted the nephrin protein reduction induced by HG treatment. Taken together, our results suggest that enhanced signaling via an Orai1/SOCE/Calpain axis contributes to HG-induced podocyte injury.     

Environmental filtering rather than dispersal limitation dominated plant community assembly in the Zoige Plateau

Identifying the mechanisms that underlie the assembly of plant communities is critical to the conservation of terrestrial biodiversity. However, it is seldom measured or quantified how much deterministic versus stochastic processes contribute to community assembly in alpine meadows. Here, we measured the decay in community similarity with spatial and environmental distance in the Zoige Plateau. Furthermore, we used redundancy analysis (RDA) to divide the variations in the relative abundance of plant families into four components to assess the effects of environmental and spatial. Species assemblage similarity liner declined with geographical distance (p < .001, R ² = .6388), and it decreased significantly with increasing distance of total phosphorus (TP), alkali‐hydrolyzable nitrogen (AN), available potassium (AK), nitrate nitrogen (NO₃ ⁺–N), and ammonia nitrogen (NH₄ ⁺–N). Environmental and spatial variables jointly explained a large proportion (55.2%) of the variation in the relative abundance of plant families. Environmental variables accounted for 13.1% of the total variation, whereas spatial variables accounted for 11.4%, perhaps due to the pronounced abiotic gradients in the alpine areas. Our study highlights the mechanism of plant community assembly in the alpine ecosystem, where environmental filtering plays a more important role than dispersal limitation. In addition, a reasonably controlled abundance of Compositae (the family with the highest niche breadth and large niche overlap value with Gramineae and Cyperaceae) was expected to maintain sustainable development in pastoral production. These results suggest that management measures should be developed with the goal of improving or maintaining suitable local environmental conditions.     

森林景观变化的地理环境因素整合分析——以伊洛河流域洛宁县为例

运用景观生态学的基本原理,借助于地理信息系统技术,整合分析了地理环境因素对伊洛河流域洛宁县森林景观动态变化的影响。结果显示,应用地理环境因素影响指数可以很好地从整体上分析地理环境因素,如高程、坡度、距离居民点中心的距离、与森林边缘距离等地理环境因子对区域森林景观动态变化的影响。1983和1999年,伊洛河流域洛宁县森林景观在地理环境因素影响指数上的优势分布区间(Pie1)分别为1230和1030,即在中高地理环境因素影响指数分布区间内,且有向低地理环境因素影响指数分布区间略为移动的趋势。森林景观类型保持不变部分、森林景观转化为非森林景观类型部分和非森林景观转化为森林景观类型部分所对应的在地理环境因素影响指数上的优势分布区间(Pie1)分别为1130、18和613。通过建立森林景观动态变化与地理环境因素影响指数的相互关系,表明森林景观动态变化与地理环境因素影响指数(特别是在地理环境因素影响指数的优势分布区间内)有显著相关关系。     

Regulation of progesterone during follicular development by FSH and LH in sheep

Progesterone (P4) can participate in the development of female mammalian antral follicles through nuclear receptor (PGR). In this experiment, the differences of P4 synthesis and PGR expression in different developmental stages of sheep antral follicles (large > 5mm, medium 2-5mm, small < 2mm) were detected by enzyme-linked immunosorbent assay, immunohistochemistry, qRT-PCR and Western blotting. Secondly, sheep follicular granulosa cells were cultured in vitro. The effects of different concentrations of FSH and LH on P4 synthesis and PGR expression were studied. The results showed that acute steroid regulatory protein (StAR), cholesterol side chain lyase (P450scc) and 3β Hydroxysteroid dehydrogenase (3β-HSD) and PGR were expressed in antral follicles, and with the development of antral follicles in sheep, StAR, P450scc and the expression of 3β-HSD and PGR increased significantly. In vitro experiments showed that FSH and LH alone or together treatment could regulate P4 secretion and PGR expression in sheep follicular granulosa cells to varying degrees, hint P4 and PGR by FSH and LH, and LH was the main factor. Our results supplement the effects of FSH and LH on the regulation of P4 synthesis during follicular development, which provides new data for further study of steroid synthesis and function in follicular development.     

Direct adenylation from 5′-OH-terminated oligonucleotides by a fusion enzyme containing Pfu RNA ligase and T4 polynucleotide kinase

5′-Adenylated oligonucleotides (AppOligos) are widely used for single-stranded DNA/RNA ligation in next-generation sequencing (NGS) applications such as microRNA (miRNA) profiling. The ligation between an AppOligo adapter and target molecules (such as miRNA) no longer requires ATP, thereby minimizing potential self-ligations and simplifying library preparation procedures. AppOligos can be produced by chemical synthesis or enzymatic modification. However, adenylation via chemical synthesis is inefficient and expensive, while enzymatic modification requires pre-phosphorylated substrate and additional purification. Here we cloned and characterized the Pfu RNA ligase encoded by the PF0353 gene in the hyperthermophilic archaea Pyrococcus furiosus. We further engineered fusion enzymes containing both Pfu RNA ligase and T4 polynucleotide kinase. One fusion enzyme, 8H-AP, was thermostable and can directly catalyze 5′-OH-terminated DNA substrates to adenylated products. The newly discovered Pfu RNA ligase and the engineered fusion enzyme may be useful tools for applications using AppOligos.     

快速检测猪肉中单核细胞增多症李氏菌PCR试剂盒的研制

快速检测猪肉中单核细胞增多症李氏菌的ＰＣＲ试剂盒与分离培养法平行检测１５６份猪肉样品,结果ＰＣＲ的阳性率为５．８％,分离培养法的阳性率为３．８％,两者符合率为９７％。实验证明,该试剂盒具有良好的特异性和可重复性,可测出样品中至少３２ＯＣＦＵ细菌,并在２ｄ内取得结果,具有推广应用价值。     

应用免疫金银技术检测猪瘟病毒兔化弱毒株感染细胞

在应用免疫金银染色技术成功地对猪瘟病毒弱毒疫苗Thiveral株（CSFV．T株）感染的PK－15细胞进行检测的基础上,通过改进免疫金银反应的条件,进一步提高了免疫金银法的检测灵敏度和特异性。目前已将该技术应用到对猪瘟病毒兔化弱毒疫苗株（CSFVC株）感染细胞的检测,获得较为满意的实验结果。免疫金银染色技术的应用为研究CSFV在细胞中增殖的规律提供更为灵敏的检测手段,并且可望为猪瘟疫苗生产中的滴度测定提供一种简便有效的手段。