Characterization of Cre recombinase mouse lines enabling cell type-specific targeting of postnatal intervertebral discs

Cre/loxP technology is an important tool for studying cell type-specific gene functions. Cre recombinase mouse lines, including Agc1-CreER^T2, Col2a1-Cre; Col2a1-CreER^T2, Shh-Cre, Shh-CreER^T2, and Osx-Cre, have been proven to be valuable tools to elucidate the biology of long bones, yet the information for their activity in postnatal intervertebral disc (IVD) tissues was very limited. In this study, we used R26-mTmG fluorescent reporter to systematically analyze cell specificity and targeting efficiency of these six mouse lines in IVD tissues at postnatal growing and adult stages. We found that Agc1-CreER^T2 is effective to direct recombination in all components of IVDs, including annulus fibrosus (AF), nucleus pulposus (NP), and cartilaginous endplate (CEP), upon tamoxifen induction at either 2 weeks or 2 months of ages. Moreover, Col2a1-Cre targets most of the cells in IVDs, except for some cells in the outer AF (OAF) and NP. In contrast, the activity of Col2a1-CreER^T2 is mainly limited to the IAF of IVD tissues at either stage of tamoxifen injection. Similarly, Shh-Cre directs recombination specifically in all NP cells, whereas Shh-CreER^T2 is active only in a few NP cells when tamoxifen is administered at either stage. Finally, Osx-Cre targets cells in the CEP, but not in the NP or AF of IVDs tissues at these two stages. Thus, our data demonstrated that all these Cre lines can direct recombination in IVD tissues at postnatal stages with different cell type specificity and/or targeting efficiency, and can, therefore, serve as valuable tools to dissect cell type-specific gene functions in IVD development and homeostasis.     

Dynamic Neural Network of Insight: A Functional Magnetic Resonance Imaging Study on Solving Chinese ‘Chengyu’ Riddles

The key components of insight include breaking mental sets and forming the novel, task-related associations. The majority of researchers have agreed that the anterior cingulate cortex may mediate processes of breaking one’s mental set, while the exact neural correlates of forming novel associations are still debatable. In the present study, we used a paradigm of answer selection to explore brain activations of insight by using event-related functional magnetic resonance imaging during solving Chinese ‘chengyu’ (in Chinese pinyin) riddles. Based on the participant’s choice, the trials were classified into the insight and non-insight conditions. Both stimulus-locked and response-locked analyses are conducted to detect the neural activity corresponding to the early and late periods of insight solution, respectively. Our data indicate that the early period of insight solution shows more activation in the middle temporal gyrus, the middle frontal gyrus and the anterior cingulate cortex. These activities might be associated to the extensive semantic processing, as well as detecting and resolving cognitive conflicts. In contrast, the late period of insight solution produced increased activities in the hippocampus and the amygdala, possibly reflecting the forming of novel association and the concomitant “Aha” feeling. Our study supports the key role of hippocampus in forming novel associations, and indicates a dynamic neural network during insight solution.     

Characterization of the prokaryotic diversity through a stratigraphic permafrost core profile from the Qinghai-Tibet Plateau

Permafrost on the Qinghai-Tibet Plateau is one of the most sensitive regions to climate warming, thus characterizing its microbial diversity and community composition may be important for understanding their potential responses to climate changes. Here, we investigated the prokaryotic diversity in a 10-m-long permafrost core from the Qinghai-Tibet Plateau by restriction fragment length polymorphism analysis targeting the 16S rRNA gene. We detected 191 and 17 bacterial and archaeal phylotypes representing 14 and 2 distinct phyla, respectively. Proteobacteria was the dominant bacterial phylum, while archaeal communities were characterized by a preponderance of Thaumarchaeota. Some of prokaryotic phylotypes were closely related to characterized species involved in carbon and nitrogen cycles, including nitrogen fixation, methane oxidation and nitrification. However, the majority of the phylotypes were only distantly related to known taxa at order or species level, suggesting the potential of novel diversity. Additionally, both bacterial α diversity and community composition changed significantly with sampling depth, where these communities mainly distributed according to core horizons. Arthrobacter-related phylotypes presented at high relative abundance in two active layer soils, while the deeper permafrost soils were dominated by Psychrobacter-related clones. Changes in bacterial community composition were correlated with most measured soil variables, such as carbon and nitrogen contents, pH, and conductivity.     

Pyrosequencing Investigation into the Bacterial Community in Permafrost Soils along the China-Russia Crude Oil Pipeline (CRCOP)

The China-Russia Crude Oil Pipeline (CRCOP) goes through 441 km permafrost soils in northeastern China. The bioremediation in case of oil spills is a major concern. So far, little is known about the indigenous bacteria inhabiting in the permafrost soils along the pipeline. A pilot 454 pyrosequencing analysis on the communities from four selected sites which possess high environment risk along the CRCOP is herein presented. The results reveal an immense bacterial diversity than previously anticipated. A total of 14448 OTUs with 84834 reads are identified, which could be assigned into 39 different phyla, and 223 families or 386 genera. Only five phyla sustain a mean OTU abundance more than 5% in all the samples, but they altogether account for 85.08% of total reads. Proteobacteria accounts for 41.65% of the total OTUs or 45% of the reads across all samples, and its proportion generally increases with soil depth, but OTUs numerically decline. Among Proteobacteria, the abundance of Beta-, Alpha-, Delta- and Gamma- subdivisions average to 38.7% (2331 OTUs), 37.5% (2257 OTUs), 10.35% (616 OTUs), and 6.21% (374 OTUs), respectively. Acidobacteria (esp. Acidobacteriaceae), Actinobacteria (esp. Intrasporangiaceae), Bacteroidetes (esp. Sphingobacteria and Flavobacteria) and Chloroflexi (esp. Anaerolineaceae) are also very common, accounting for 8.56% (1237 OTUs), 7.86% (1136 OTUs); 7.35% (1063 OTUs) and 8.27% (1195 OTUs) of total libraries, respectively. The ordination analysis indicates that bacteria communities in the upper active layer cluster together (similar), while bacterial consortia from the lower active layer and permafrost table scatter (less similar). The abundance of Rhodococcus (12 OTUs), Pseudomonas (71 OTUs) and Sphingomonas (87 OTUs) is even less (<0.01%). This effort to profile the background diversity may set the first stage for better evaluating the bacterial dynamics in response to accidental oil spills.     

化妆品中耐热大肠菌群和金黄色葡萄球菌检测能力验证分析

为增强实验室竞争力,考察实验室检测水平,本实验室参加了ACAS-pT743(2019)化妆品耐热大肠菌群及金黄色葡萄球菌的检测能力验证.本次能力验证中实验室共接收了四个样品,按照《化妆品安全技术规范》2015版对样品展开检测,并采用关键生理生化指标对可疑菌落加以确定以及16 S rDNA序列测定方法对结果进行复确认.结...     

The effect and implication of human disturbances on altitudinal variation of non-structural carbohydrates in Kobresia pygmaea

Non-structural carbohydrates in plant organs can mirror the plant overall carbon supply status and balance and can also provide evidence for their health evaluation in the ecosystem. Non-structural carbohydrates in Kobresia pygmaea, one dominant herbaceous species on the Qinghai-Tibetan Plateau, were measured to investigate altitudinal variation in non-structural carbohydrate accumulations, as well as engineering disturbance on the stability of the alpine grassland ecosystem. An increasing trend with elevation in total soluble sugars, fructose, and sucrose was detected in the K. pygmaea growing in both undisturbed and disturbed sites. However, there were higher amounts and a more distinctly altitudinal trend of non-structural carbohydrates with a minor fluctuation in undisturbed sites compared to disturbed sites. In addition, the altitudinal trend of sucrose is similar to that in sucrose phosphate synthase and sucrose synthase activities, while it is opposite to that in neutral invertase and soluble acid invertase activities, suggesting that the sucrose accumulation was primarily related to its synthesis. These results revealed that human disturbance resulted in a reduced carbon supply and altered the balance of carbohydrate utilization in plants on the Qinghai-Tibetan Plateau.     

Relative Roles of Deterministic and Stochastic Processes in Driving the Vertical Distribution of Bacterial Communities in a Permafrost Core from the Qinghai-Tibet Plateau,China

Understanding the processes that influence the structure of biotic communities is one of the major ecological topics, and both stochastic and deterministic processes are expected to be at work simultaneously in most communities. Here, we investigated the vertical distribution patterns of bacterial communities in a 10-m-long soil core taken within permafrost of the Qinghai-Tibet Plateau. To get a better understanding of the forces that govern these patterns, we examined the diversity and structure of bacterial communities, and the change in community composition along the vertical distance (spatial turnover) from both taxonomic and phylogenetic perspectives. Measures of taxonomic and phylogenetic beta diversity revealed that bacterial community composition changed continuously along the soil core, and showed a vertical distance-decay relationship. Multiple stepwise regression analysis suggested that bacterial alpha diversity and phylogenetic structure were strongly correlated with soil conductivity and pH but weakly correlated with depth. There was evidence that deterministic and stochastic processes collectively drived bacterial vertically-structured pattern. Bacterial communities in five soil horizons (two originated from the active layer and three from permafrost) of the permafrost core were phylogenetically random, indicator of stochastic processes. However, we found a stronger effect of deterministic processes related to soil pH, conductivity, and organic carbon content that were structuring the bacterial communities. We therefore conclude that the vertical distribution of bacterial communities was governed primarily by deterministic ecological selection, although stochastic processes were also at work. Furthermore, the strong impact of environmental conditions (for example, soil physicochemical parameters and seasonal freeze-thaw cycles) on these communities underlines the sensitivity of permafrost microorganisms to climate change and potentially subsequent permafrost thaw.     

噬菌体作为指示病毒的应用研究进展北大核心

噬菌体是一类专性侵染细菌的病毒,在形态大小、结构组成等生物特性上与高等生物病毒具有相似性,同时噬菌体实验室操作技术简单,安全性高,在培养、计数、稳定性和灵敏度等方面具有非常大的优势。因此,将噬菌体作为指示生物模拟或替代高等生物病毒的研究和应用已开始受到国内外研究人员的关注,并取得一定进展。本文论述了噬菌体作为指示病毒的优势,并对噬菌体在病毒过滤去除效果评价、消毒效果评价、病毒传播规律研究、环境及水质监测等领域的研究和应用进行了总结分析,综述了噬菌体在各领域应用的可行性证据、应用案例及难点问题,并结合噬菌体作为指示病毒的不足之处,对进一步以噬菌体作为指示病毒的研究和应用提出建议和展望。     

High carbon emissions from thermokarst lakes and their determinants in the Tibet Plateau

Thermokarst lakes are potentially important sources of methane (CH₄) and carbon dioxide (CO₂). However, considerable uncertainty exists regarding carbon emissions from thermokarst lakes owing to a limited understanding of their patterns and motivators. In this study, we measured CH₄ and CO₂ diffusive fluxes in 163 thermokarst lakes in the Qinghai–Tibet Plateau (QTP) over 3 years from May to October. The median carbon emissions from the QTP thermokarst lakes were 1440 mg CO₂ m⁻² day⁻¹ and 60 mg CH₄ m⁻² day⁻¹, respectively. The diffusive rates of CO₂ and CH₄ are related to the catchment land cover type. Sediment microbial abundance and hydrochemistry explain 51.9% and 38.3% of the total variance in CH₄ diffusive emissions, respectively, while CO₂ emissions show no significant relationship with environmental factors. When upscaling carbon emissions from the QTP thermokarst lakes, the annual average CH₄ release per lake area is equal to that of the pan-Arctic region. Our findings highlight the importance of incorporating in situ observation data with different emission pathways for different land cover types in predicting carbon emissions from thermokarst lakes in the future.