Engineering the mouse genome with bacterial artificial chromosomes to create multipurpose alleles

The mouse is the leading vertebrate model because its genome can be altered by both random transgenesis and homologous recombination with targeting constructs. Both approaches have been hindered by the size and site limitations implicit in conventional Escherichia coli DNA-engineering methods. Homologous recombination in E. coli, or 'recombineering', has overcome these limitations for bacterial artificial chromosome (BAC) transgenesis. Here we applied Red/ET recombineering (using the lambda Redalpha/Redbeta recombinase pair) to generate a 64 kilobase targeting construct that carried two selectable cassettes permitting the simultaneous mutation of the target gene, Mll, at sites 43 kb apart in one round of mouse embryonic stem (ES) cell targeting. The targeting frequency after dual selection was 6%. Because the two selectable cassettes were flanked by FRT or loxP sites, three more alleles can be generated by site-specific recombination. Our approach represents a simple way to introduce changes at two or more sites in a genetic locus, and thereafter generate allele combinations. The size of BAC templates offers new freedom for the design of targeting constructs. Combined with the use of two selectable cassettes placed far apart, BAC-based targeting constructs may be applicable to tasks such as regional exchanges, deletions, and insertions.     

Body Mass Index Is Associated with Inflammatory Bowel Disease: A Systematic Review and Meta-Analysis

Background

Prior work suggested that patients with inflammatory bowel diseases (IBD) have lower body mass index (BMI) than controls and patients with lower BMI have more serious complications.

Goal

The study was aimed to find relationship between BMI in patients with and without IBD, investigate effects of medicine therapy and disease stages on patients’ BMI.

Methods

Potentially eligible studies were identified through searching PubMed, Cochrane and Embase databases. Outcome measurements of mean BMI and the number of patients from each study were pooled by a random-effect model. Publication bias test, sensitivity analysis and subgroup analysis were conducted.

Results

A total of 24 studies containing 1442 patients and 2059 controls were included. Main results were as follows: (1) BMI in Crohn’s disease (CD) patients was lower than that in health controls (-1.88, 95% CI -2.77 to -1.00, P< 0.001); (2) Medical therapy significantly improved BMI of CD patients (with therapy: -1.58, -3.33 to 0.16; without: -2.09, 95% CI -3.21 to -0.98) while on the contrary not significantly improving BMI of UC patients (with therapy: -0.24, 95% CI -3.68 to 3.20; without: -1.34, 95% CI -2.87 to 0.20, P = 0.57); (3) Both CD and UC patients in active phase showed significantly greater BMI difference compared with controls than those in remission (CD patients: remission: -2.25, 95% CI -3.38 to -1.11; active phase: -4.25, 95% CI -5.58 to -2.92, P = 0.03; UC patients: remission: 0.4, 95% CI -2.05 to 2.84; active phase: -5.38, -6.78 to -3.97, P = 0.001).

Conclusions

BMI is lower in CD patients; medical therapy couldn’t improve BMI of IBD patients; the state of disease affects BMI of CD patients and UC patients.     

Levofloxacin susceptibility testing for Helicobacter pylori in China: comparison of E-test and disk diffusion method

Background: The aims of this study were to compare disk diffusion with E‐test method for levofloxacin susceptibility testing of Helicobacter pylori and standardized breakpoints for disk diffusion as a stable and reliable method for determining qualitative levofloxacin susceptibility. Materials and Methods: We determined the levofloxacin susceptibility of 45 H. pylori strains isolated from Chinese patients by the E‐test method. Disk diffusion was evaluated as an alternative method to determine susceptibility and compared with the E‐test results by linear regression analysis. Results: The minimum inhibitory concentration (MIC) values tested by E‐test method ranged from 0.047 to 32 μg/mL. Resistance to levofloxacin was detected in 16 (35.6%) isolates. The levofloxacin disk zone sizes obtained by disk diffusion method correlated well (r² = .877) with the MICs obtained by E‐test method. As a consequence of regression analysis, isolates with inhibition diameters <12 mm were considered resistant to levofloxacin. There was 100% agreement between the two methods for levofloxacin, applying the regression‐based breakpoints. Conclusions: The disk diffusion method is equivalent to the E‐test method for testing levofloxacin susceptibility of H. pylori strains; it is more practical and inexpensive, and it is suitable for the analysis of a small number of isolates compared with the E‐test method.     

毛竹林冠层与林下层节肢动物类群的关系

根据生境的异质程度将毛竹林节肢动物群落划分为竹冠层和林下层类群。通过大面积系统调查,结果表明:竹冠层与林下层类群物种组成相似性高,两类群的共有种为258种,其中植食性、蜘蛛类、寄生性、捕食性昆虫和螨类、中性物种分别有88、53、41、38种和38种,主要隶属于蜘蛛目、膜翅目、双翅目和鞘翅目,天敌总共有种为132种,占共有种总数的51.16%。物种在二类群间可相互交流,林下层类群物种更替比竹冠层类群更为频繁,中性物种交流比其它类物种更为频繁,一年中3~7月份和10~11月份的物种交流多。竹冠层类群在林下层类群的重建和瓦解中起着物种“源”和“库”的作用。典型相关分析表明,二类群的关系主要表现为林下层类群丰富度和物种多样性与竹冠层类群物种多样性、均匀度、丰富度之间的关系,保护林下层天敌和物种多样性有利于保护竹冠层天敌和物种多样性。     

土地利用转变与海拔高度协同作用黄土高原植被固碳变化特征

全球固碳释碳问题一直是近年来关乎民生的热点话题,区域碳源/碳汇对生态环境的重要性不言而喻。基于CASA模型估算黄土高原1990—2015年植被净初级生产力的年际变化,并分析土地利用变化、海拔高度及两者协同作用对其综合影响,结果表明：(1)黄土高原1990—2015年植被NPP与植被固碳总体呈增加趋势,年均NPP增速2.74 gC m^-2 a^-1,年均固碳增速1.13 TgC/a,研究区林地年均NPP(619.5 gC m^-2 a^-1)远超其他用地类型,固碳效果理想;(2)黄土高原年均NPP随高程的增加先降低后升高,年总NPP和固碳量随高程增加变化趋势相反;(3)研究区土地利用转变类型中退耕还林的植被固碳效果最好;而林地变为耕地或草地均不能达到固碳目的,此外,更推荐在研究区海拔低于1500 m变草为耕,海拔高于1500 m退耕还草,海拔高于3000 m变耕、草为林。以期为区域尺度的生态环境建设提供一定的参考和科学依据。     

Expression of functional Kir6.1 channels regulates glutamate release at CA3 synapses in generation of epileptic form of seizures

The Kir6.1 channels are a subtype of ATP-sensitive inwardly rectifying potassium (K_ATP) channels that play an essential role in coupling the cell's metabolic events to electrical activity. In this study, we show that functional Kir6.1 channels are located at excitatory pre-synaptic terminals as a complex with type-1 Sulfonylurea receptors (SUR1) in the hippocampus. The mutant mice with deficiencies in expressing the Kir6.1 or the SUR1 gene are more vulnerable to generation of epileptic form of seizures, compared to wild-type controls. Whole-cell patch clamp recordings demonstrate that genetic deletion of the Kir6.1/SUR1 channels enhances glutamate release at CA3 synapses. Hence, expression of functional Kir6.1/SUR1 channels inhibits seizure responses and possibly acts via limiting excitatory glutamate release.     

Differentiation of liver cells from human primordial germ cell-derived progenitors

In previous studies, progenitor embryoid body-derived (EBD) cells have been derived from human embryonic germ cells. These cells express lineage markers of three primary germ layers, although their potential to produce true fetal cells of various types has yet to be tested. To this end, we have transplanted EBD cells into the fetal sheep liver. We show that these cells respond appropriately to environmental cues and give rise to hepatocytes and well-structured bile ducts. These results suggest that EBD cells are relatively uncommitted early progenitors capable of effective incorporation and differentiation in vivo. The ability to generate functional liver cells makes EBD cells potentially useful for cell therapy.