Physiological properties and classification of strains of Treponema sp. isolated from pigs in poland

Fifty-one treponemas were isolated from pigs. Twenty-three isolates with typical morphology and growth characteristic were beta hemolytic, enteropathogenic, produced indole and with exception of three strains did not ferment fructose. These strains were classified as typical T. hyodysenteriae and were usually isolated from pigs with symptoms of mucohemorrhagic diarrhoea. The seventeen other isolates were weakly beta hemolytic after 48 h incubation, enteropathogenic, 12 out of 17 produced indole, 10 out 17 fermented fructose. These strains were usually isolated from pigs with symptoms of gray-green diarrhoea and classified as T. hyodysenteriae 2 biotype or intermediate type. They may be compared with Treponema sp. isolated by Taylor et al. Eleven non enteropathogenic strains showed typical characteristic for T. innocens. Gas chromatography analysis of the fatty acids production from glucose, showed that all isolated treponemas produced acetate and butyrate. Typical T. hyodysenteriae produced additionally propionate. Strains of T. hyodysenteriae biotype 2 produced propionate or isobutyrate as well.     

CpG-ODN enhances mammary gland defense during mastitis induced by Escherichia coli infection in goats

Seven healthy native goats in early lactation, weighing 30–40 kg, were used in this study. The right mammary gland of the seven does were infused with CpG-ODN at a dosage of 100 μg kg⁻¹ body weight on the day 5 postpartum (PP). The left glands were used as controls and infused with sterile phosphate-buffered saline (PBS). On day 8 PP, the same dosage of CpG-ODN or PBS was again infused. On day 9 PP, the mammary glands (both right and left) of the seven does were infused with 6 × 10⁶ colony-forming units (CFU) Escherichia coli and, at 0, 8, 16, 24, 48 and 72 h postinfection (PI), milk samples were collected from all glands. Goats were euthanized at 72 h PI and the mammary tissue harvested. Infusion with 6 × 10⁶ CFU ml⁻¹ E. coli induced acute mastitis. Histopathological evaluations showed that polymorphonuclear neutrophils (PMNs) were still present in alveoli at 72 h PI, but PMNs in the CpG-ODN-treated glands has disappeared. Bacteria counts in milk peaked at 16 h PI and CpG-ODN induced a significant decrease in viable bacteria from 16 h PI until the end of the experiment. This study showed that CpG-ODN promoted the expression of its specific receptor (TLR-9 mRNA) in mammary tissue, stimulated IL-6 production, reduced bacteria counts in milk, attenuated the impact of inflammation mediators on cells and significantly shortened the inflammation course. These results suggest that the CpG-ODN improved mammary gland defense and, thereby, had a beneficial effects against mastitis caused by E. coli infection in goats.     

基于CRISPR/Cas9技术建立IGF2R基因修饰PK-15细胞

作为我国特色的地方种质资源,藏猪对高海拔、低氧、强紫外线辐射等恶劣环境具有良好的适应性,前期研究发现藏猪的IGF2R（insulin-like growth factor 2 receptor,IGF2R）基因在其内含子上有1段274 bp的缺失,且该缺失为藏猪独有的变异,关于IGF2R是否参与藏猪对低氧等恶劣环境的适应性调节的相关研究尚未报道。为了建立IGF2R基因274 bp修饰的猪细胞系,研究IGF2R基因对藏猪低氧耐受特性的影响,通过CRISPR/Cas9技术建立基因修饰PK-15细胞系,并通过PCR、实时荧光定量PCR（real time quantitative PCR,RT-qPCR）、蛋白免疫印迹（Western-blot）、免疫荧光（immunofluorescence,IF）等方法鉴定其分子特征,随后分别利用CCK-8和RT-qPCR 、Western-blot对基因修饰细胞进行活力和凋亡基因检测。结果显示,已成功构建IGF2R基因274 bp修饰的猪细胞系,IGF2R基因表达量极显著下调（P < 0.01）。基因修饰细胞活力在72 h时极显著上升（P < 0.01）,并且含半胱氨酸的天冬氨酸蛋白水解酶9 （cysteinyl aspartate specific proteinase,Caspase9）和BCL-2关联 X （BCL2-associated X,BAX） 蛋白的水平显著下调（P < 0.05）,并且B淋巴细胞瘤-2（B-cell lymphoma-2,BCL2）基因的mRNA水平极显著上升（P < 0.01）。成功构建了IGF2R基因精确修饰的PK-15细胞系,并初步进行了功能研究,为研究IGF2R基因功能提供细胞模型,并为后续基因编辑猪的制备奠定基础。     

油研9号氮、钾、硼肥的施用技术

为了探讨油菜良种良法配套栽培技术,以新品种油研9号进行了氮肥的不同施用比例、钾肥的不同施用量、硼肥的不同施用方式的研究。结果表明:在相同施氮量下,以基肥:苗肥:薹肥为3:4:3时产量最高,与5:3:2和6:2:2 的施用比例达显著性差异;与4:3:3的施用比例差异不显著。不同钾肥施用量以底施氯化钾15 kg／667m2产量最高,底施氯化钾均与不施氯化钾达极显著性差异,但底施氯化钾15 kg／667m2比底施氯化钾7．5 kg／667m2增产不显著。不同硼肥的施用方式,以薹施硼肥1 kg／667m2产量最高,薹施(1kg／667m2)、底施(1 kg／667m2)、花喷(100g／667m2)硼肥分别比不施硼肥增产 31．5％、22．8％、5．5％,施用硼肥与不施硼肥均达显著性差异,薹施和底施硼肥增产效果明显,花喷增产效果较差。     

2010-2011年河南省部分地区H9亚型禽流感、新城疫

为了解河南省H9亚型禽流感、新城疫和鸡传染性支气管炎的流行情况。从河南省15个地市发病鸡场采集病鸡组织样品309份,采用鸡胚尿囊腔传代接种法分离病毒,并利用血凝试验(HA)、血凝抑制试验(HI)和RT-PCR试验对分离毒株进行鉴定。结果表明,3类病毒的总检出率为26.54%,其中NDV、IBV和AIV(H9)的检出率分别为11.00％、3.88％和11.65％。2010年10月NDV和IBV检出率均最高,分别为30.00％、15.00％,2011年1月AIV(H9)检出率最高,为43.24％。2类病毒混合感染的检出率达3.24％。表明2010年7月-2011年4月H9亚型禽流感、新城疫以及鸡传染性支气管炎在河南省部分地区普遍流行,且存在一定程度的混合感染。为有效预防和控制河南省3种主要疫病提供了科学依据。     

MytiLec,a Mussel R-Type Lectin,Interacts with Surface Glycan Gb3 on Burkitt’s Lymphoma Cells to Trigger Apoptosis through Multiple Pathways

MytiLec; a novel lectin isolated from the Mediterranean mussel (Mytilus galloprovincialis); shows strong binding affinity to globotriose (Gb3: Galα1-4Galβ1-4Glc). MytiLec revealed β-trefoil folding as also found in the ricin B-subunit type (R-type) lectin family, although the amino acid sequences were quite different. Classification of R-type lectin family members therefore needs to be based on conformation as well as on primary structure. MytiLec specifically killed Burkitt''s lymphoma Ramos cells, which express Gb3. Fluorescein-labeling assay revealed that MytiLec was incorporated inside the cells. MytiLec treatment of Ramos cells resulted in activation of both classical MAPK/ extracellular signal-regulated kinase and extracellular signal-regulated kinase (MEK-ERK) and stress-activated (p38 kinase and JNK) Mitogen-activated protein kinases (MAPK) pathways. In the cells, MytiLec treatment triggered expression of tumor necrosis factor (TNF)-α (a ligand of death receptor-dependent apoptosis) and activation of mitochondria-controlling caspase-9 (initiator caspase) and caspase-3 (activator caspase). Experiments using the specific MEK inhibitor U0126 showed that MytiLec-induced phosphorylation of the MEK-ERK pathway up-regulated expression of the cyclin-dependent kinase inhibitor p21, leading to cell cycle arrest and TNF-α production. Activation of caspase-3 by MytiLec appeared to be regulated by multiple different pathways. Our findings, taken together, indicate that the novel R-type lectin MytiLec initiates programmed cell death of Burkitt’s lymphoma cells through multiple pathways (MAPK cascade, death receptor signaling; caspase activation) based on interaction of the lectin with Gb3-containing glycosphingolipid-enriched microdomains on the cell surface.     

敲除TMS5基因获得温敏不育粳稻新材料

【目的】水稻温敏不育系的选育是两系杂交稻育种工作中的关键环节。TMS5是控制温敏不育的重要基因,在生产上也应用较广。为了加快水稻温敏不育系的选育进程,【方法】利用CRISPR/Cas9技术,在武运粳7号背景下对TMS5基因进行编辑。【结果】通过测序从22株T0代中筛选获得6株纯合的突变体,分析发现6个纯合的突变体产生了相同的遗传变异,都在TMS5基因第2外显子44 bp和45 bp之间插入碱基"A",导致翻译提前终止。通过细胞学观察和人工辅助授粉证实获得的株系花粉败育而雌配子正常发育。【结论】这些结果表明对温敏不育基因TMS5基因进行编辑可以快速获得粳型温敏雄性不育系。     

玉米转ABP9基因植株的获得

ABP9参与ABA信号传导及ROS代谢调节,增强植物抵抗非生物逆境胁迫的能力,因此该基因在作物抗逆性改良上有潜在应用价值。为了获得转ABP9基因的玉米植株,向玉米抗逆育种提供新材料,构建了由组成型启动子Ubi驱动抗逆转录因子ABP9基因的植物表达载体,并通过农杆菌介导对玉米自交系齐319进行了遗传转化。经草铵膦抗性筛选和用两对不同的基因特异引物进行PCR扩增检测,证明已将ABP9基因导入玉米齐319基因组中,获得转基因植株。     

不同施磷量对油菜新品种产量的影响

通过田间试验,研究磷肥与油菜产量、品质和植株形态之间的关系,确定油菜高产、优质、高效的磷肥施用量。不同油菜品种磷肥用量试验的结果表明:①每公顷施纯磷360kg的处理,中双9号,每公顷产量达3301.65kg,中油杂2号,产量达到最大值,每公顷产量3781.95kg。②一次分枝对产量影响最大,二次分枝、主花序的贡献相对要小得多。③随着施磷量的增加单位面积有效角果数呈增长的趋势。     

B9和6-BA配合使用对唐菖蒲切花保鲜效果影响的研究

[目的]为提高唐菖蒲切花的保鲜效果提供理论依据及技术指导。[方法]以粉秀唐菖蒲品种为供试材料,通过对唐菖蒲寿命、观赏值、水分平衡值、鲜重变化率、花青素含量、花瓣质膜相对透性等指标的观测,探讨了B9和6-苄基嘌呤(6-BA)的不同组合对唐菖蒲切花保鲜效果的影响。[结果]各处理切花的瓶插寿命均明显长于CK,而添加6-BA和B9的处理其瓶插寿命与母液相比有较明显的延长。6-BA与B9间存在明显的交互作用。6-BA在增加唐菖蒲切花鲜重,改善切花体内的水分状况等方面的作用效果比B9要好。各保鲜剂处理均可抑制植物体内膜脂过氧化物丙二醛的产生,维持细胞膜结构的稳定性。各处理花青素含量随瓶插天数的增加呈先上升后下降的趋势。[结论]综合考虑,以4%蔗糖+300mg/L8-羟基喹啉+150mg/L硼酸+20mg/L6-BA+200mg/LB9的保鲜效果最佳,瓶插寿命达到22d。