Efficient GFP‐labeling and analysis of spermatogenic cells using the IRG transgene and flow cytometry

Spermatogenesis is a highly ordered developmental program that produces haploid male germ cells. The study of male germ cell development in the mouse has provided unique perspectives into the molecular mechanisms that control cell development and differentiation in mammals, including tissue‐specific gene regulatory programs. An intrinsic challenge in spermatogenesis research is the heterogeneity of germ and somatic cell types present in the testis. Techniques to separate and isolate distinct mouse spermatogenic cell types have great potential to shed light on molecular mechanisms controlling mammalian cell development, while also providing new insights into cellular events important for human reproductive health. Here, we detail a versatile strategy that combines Cre‐lox technology to fluorescently label germ cells, with flow cytometry to discriminate and isolate germ cells in different stages of development for cellular and molecular analyses.     

Effect of tannery sludge amendments on the activity of soil enzymes and phytoremediation potential of two economically important cultivars of geranium (Pelargonium graveolens)

A field experiment was conducted to observe the effect of TS amendments on soil enzymes and phytoremediation potential of two economically important cultivars of geranium. Different doses of TS were applied in soil to examine threshold limit of HMs where geranium cultivars can be grown successfully in contaminated sites. Treatment variation significantly affected pH, EC, OC, N, P, K and HM content in soil after 50 days of incubation. After harvest, both cultivars were examined to assess the impact of various treatments on their fresh herb, dry matter, essential oil yield and HM accumulation. C/G ratio close to 1 was observed at 50 tha^?1 sludge treatment in both cultivars. Urease and β-glucosidase activities in soil were maximum at 50 tha^?1 whereas dehydrogenase and phosphatase activities were maximum at 100 tha^?1 in both cultivars. β-glucosidase, acid and alkaline phosphatase, urease and dehydrogenase activities were relatively high after 85 days over 45 days in both cultivars. Maximum metal uptake was found in roots of cv. Bourbon followed by leaves. Geranium was observed to be a good candidate for phytoremediation as it mitigates metal toxicity by root absorption and cv. Bourbon is better candidate for the same.     

The Middle/Later Stone Age transition and cultural dynamics of late Pleistocene East Africa

The Middle to Later Stone Age (MSA/LSA) transition is a prominent feature of the African archeological record that began in some places ~30,000–60,000 years ago, historically associated with the origin and/or dispersal of “modern” humans. Unlike the analogous Middle to Upper Paleolithic transition in Eurasia and associated Neanderthal extinction, the African MSA/LSA record remains poorly documented, with its potential role in explaining changes in the behavioral diversity and geographic range of Homo sapiens largely unexplored. I review archeological and biogeographic data from East Africa, show regionally diverse pathways to the MSA/LSA transition, and emphasize the need for analytical approaches that document potential ancestor‐descendent relationships visible in the archeological record, needed to assess independent invention, population interaction, dispersal, and other potential mechanisms for behavioral change. Diversity within East Africa underscores the need for regional, rather than continental‐scale narratives of the later evolutionary history of H. sapiens.     

基因敲除技术及其在研究线粒体动力学与胰岛素抵抗关系中的应用进展

线粒体动力学即线粒体融合和分裂保持动态平衡的过程,该过程由融合/分裂相关蛋白精确调控完成,对于线粒体代谢、质量和功能有着重要的生理意义,而这些蛋白发生异常可引发线粒体动力学失衡,进而引起线粒体功能障碍并引发各种疾病状态。文中围绕基因敲除技术,详细阐述了编码融合/分裂相关蛋白的基因敲除鼠在胰岛素抵抗研究工作中的作用及应用进展,以期为今后研究线粒体动力学失衡致胰岛素抵抗的信号转导机制奠定基础。     

微生物菌种保藏方法及关键技术

菌种是国家的重要生物资源,也是生产、教学和科学研究的基本材料。为确保菌种的质量和活力,需要正确的菌种保藏方法。阐述了菌种保藏的重要性、菌种保藏常见方法及其存在的问题,探讨了常用菌种的保藏技术及关键点,好的保藏方法可延长菌种的保存时间,又可防止菌种退化。     

Second and third harmonic generation microscopy visualizes key structural components in fresh unprocessed healthy human breast tissue

Real‐time assessment of excised tissue may help to improve surgical results in breast tumor surgeries. Here, as a step towards this purpose, the potential of second and third harmonic generation (SHG, THG) microscopy is explored. SHG and THG are nonlinear optical microscopic techniques that do not require labeling of tissue to generate 3D images with intrinsic depth‐sectioning at sub‐cellular resolution. Until now, this technique had been applied on fixated breast tissue or to visualize the stroma only, whereas most tumors start in the lobules and ducts. Here, SHG/THG images of freshly excised unprocessed healthy human tissue are shown to reveal key breast components—lobules, ducts, fat tissue, connective tissue and blood vessels, in good agreement with hematoxylin and eosin histology. DNA staining of fresh unprocessed mouse breast tissue was performed to aid in the identification of cell nuclei in label‐free THG images. Furthermore, 2‐ and 3‐photon excited auto‐fluorescence images of mouse and human tissue are collected for comparison. The SHG/THG imaging modalities generate high quality images of freshly excised tissue in less than a minute with an information content comparable to that of the gold standard, histopathology. Therefore, SHG/THG microscopy is a promising tool for real‐time assessment of excised tissue during surgery.      

CRISPR/Cas技术在抗除草剂作物育种中的研究与应用进展

CRISPR/Cas系统是一种简单、低成本、高效、精准的基因编辑技术,该技术能够进行基因的定向改造,加速新品种培育进程,在种质资源创制中的应用潜力较高。概述了CRISPR/Cas系统的技术原理及其在作物抗除草剂育种中的应用,简要指出了目前CRISPR/Cas技术在抗除草剂种质创制及应用过程中存在的问题及发展方向,以期为今后利用CRISPR/Cas技术创制抗除草剂新种质提供理论依据。     

Preparation and application of freeze-dried platelets

Clinical platelet infusion is primarily used to prevent or stop bleeding, but can also have a role in treating infections or promoting wound healing. The demand for platelets has increased in recent years. However, as platelets can only be stored for short periods, there is a substantial loss due to the products reaching their expiry date. Platelet lyophilization is a particularly valuable and important research field. The purpose of studying the freeze-drying preservation of platelets is to realize the long-term preservation of platelets at room temperature. It is very possible to prepare qualified freeze-dried platelets. However, there are still problems that have not been solved in the process of platelet lyophilization. This review mainly summarizes research progress in the preparation and application of freeze-dried platelets.     

Expression of bacterial biphenyl-chlorobiphenyl dioxygenase genes in tobacco plants

Optimized plant-microbe bioremediation processes in which the plant initiates the metabolism of xenobiotics and releases the metabolites in the rhizosphere to be further degraded by the rhizobacteria is a promising alternative to restore contaminated sites in situ. However, such processes require that plants produce the metabolites that bacteria can readily oxidize. The biphenyl dioxygenase is the first enzyme of the bacterial catabolic pathway involved in the degradation of polychlorinated biphenyls. This enzyme consists of three components: the two sub-unit oxygenase (BphAE) containing a Rieske-type iron-sulfur cluster and a mononuclear iron center, the Rieske-type ferredoxin (BphF), and the FAD-containing ferredoxin reductase (BphG). In this work, based on analyses with Nicotiana benthamiana plants transiently expressing the biphenyl dioxygenase genes from Burkholderia xenovorans LB400 and transgenic Nicotiana tabacum plants transformed with each of these four genes, we have shown that each of the three biphenyl dioxygenase components can be produced individually as active protein in tobacco plants. Therefore, when BphAE, BphF, and BphG purified from plant were used to catalyze the oxygenation of 4-chlorobiphenyl, detectable amounts of 2,3-dihydro-2, 3-dihydroxy-4'-chlorobiphenyl were produced. This suggests that creating transgenic plants expressing simultaneously all four genes required to produce active biphenyl dioxygenase is feasible.     

Availability and contributions of soil phosphorus to forage production of seeded alfalfa in semiarid Loess Plateau

The availability of soil phosphorus was studied in response to the forage production of seeded alfalfa (Medicago sativa L.), established by the ridge and furrow water-harvesting technique in the semiarid Loess Plateau of China. The following 5 treatments were set up in this study: (1) conventional cultivation in a flat plot without mulch (CK); (2) plastic mulched ridge with 30 cm width of ridge and furrow (M30); (3) plastic mulched ridge with 60 cm width of ridge and furrow (M60); (4) bare ridge with 30 cm width of ridge and furrow (B30); (5) bare ridge with 60 cm width of ridge and furrow (B60). The forage yield, evapotranspiration (ET), soil organic carbon (SOC), total phosphorus, available phosphorus and the ratio of soil organic C to available P (C/P) were measured in the experiment. Results showed that the positive correlation occurred between the decrease in soil available P and the total forage yield of alfalfa after three-year alfalfa stand. M30 and M60 were higher than those of conventional flat cultivation (CK) by 10.7% and 40.3%, respectively, whereas the yield of two bare treatments (B30 and B60) was lower than that of CK by 14.2% and 28.3%, respectively. After a 3-year alfalfa stand, the trend of the decrease in soil available P was in the order of M60 (55.5%) > M30 (51.5%) > CK (34.6%) > B30 (23.4%) > B60 (17.5%). Moreover, after 3-year alfalfa growth, the C/P ratio increased significantly in all of the treatments by comparing with that before sowing. The C/P ratios of M30 (1165.1) and M60 (1326.1) were significantly higher than those of the other treatments. More interesting finding was that the increase in SOC in dry years promoted the accumulation of soil available P. However, in wet years, the increase of SOC limited the accumulation of soil available P. Therefore, to further increase the forage yield of alfalfa, the effective technique must be developed to increase the soil available P.