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.     

The effects of ionic liquid 1-ethyl-3-methylimidazolium trifluoromethanesulfonate on the production of 1,3-propanediol from crude glycerol by microbial consortium

Bioprocess and Biosystems Engineering - Ionic liquids (ILs) as “green” solvents have been widely used owing to their excellent properties, e.g., for biodiesel production. Crude glycerol...     

Lyoniresinol 3α-O-β-D-Glucopyranoside-Mediated Hypoglycaemia and Its Influence on Apoptosis-Regulatory Protein Expression in the Injured Kidneys of Streptozotocin-Induced Mice

Averrhoa carambola L. (Oxalidaceae) root (ACLR) has a long history of use in traditional Chinese medicine for treating diabetes and diabetic nephropathy (DN). (±)-Lyoniresinol 3α-O-β-D-glucopyranoside (LGP1, LGP2) were two chiral lignan glucosides that were isolated from the ACLR. The purpose of this study was to investigate the effect of LGP1 and LGP2-mediated hypoglycaemia on renal injury in streptozotocin (STZ)-induced diabetic mice. STZ-induced diabetic mice were administrated LGP1 and LGP2 orally (20, 40, 80 mg/kg body weight/d) for 14 days. Hyperglycaemia and the expression of related proteins such as nuclear factor-κB (NF-κB), caspase-3, -8, -9, and Bcl-associated X protein (Bax) were markedly decreased by LGP1 treatment. However, LGP2 treatment had no hypoglycaemic activity. Diabetes-dependent alterations in the kidney such as glomerular hypertrophy, excessive extracellular matrix amassing, and glomerular and tubular basement membrane thickening were improved after 14 days of LGP1 treatment. B cell lymphoma Leukaemia-2 (Bcl-2) expression was reduced in the STZ-induced diabetic mouse kidneys but was enhanced by LGP1 treatment. These findings suggest that LGP1 treatment may inhibit diabetic nephropathy progression and may regulate several pharmacological targets for treating or preventing diabetic nephropathy.     

Viral tegument proteins restrict cGAS-DNA phase separation to mediate immune evasion

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蜜蜂褪黑素的测定方法

在脊椎动物中,褪黑素(5-甲氧基-N-乙酰色胺,MLT)是一种由松果体分泌的具有典型光周期信号作用的神经内分泌激素。近年在昆虫头部也发现了这种化学物质。该文简述了褪黑素常见测定方法,详细介绍了褪黑素的免疫测定方法RIA(radionimmunoassay),并使用该方法初步测定了中华蜜蜂ApisceranaFabricius工蜂头部褪黑素的含量,结果显示褪黑素含量与工蜂的社会分工相关。     

武汉东湖浮游病毒的丰度及多样性

运用透射电镜及荧光显微技术 ,对富营养化的武汉东湖中浮游病毒丰度及多样性进行了研究。电镜观察和计数结果显示 ,东湖中浮游病毒的丰度达到 10 8个 /mL。荧光显微镜计数结果约是电镜计数结果的 2 72倍 ,差异极显著 (P <0 0 1)。在东湖超富营养区的水样中观察到了多种与各种病毒形态类似的颗粒 ,其中大部分与噬菌体和噬藻体类似 ,具多种形态的尾部和六边形头部。还有一些线状、杆状、子弹形等形态的病毒粒子。研究结果显示东湖水环境中的浮游病毒不仅丰度极高 ,而且种类丰富。提示浮游病毒在东湖水环境和水生态系统中可能扮演重要角色。     

影响小鼠体细胞脂质体法转染效率的因素

We studied the effects of the amount of liposome and plasmid, exposure time of cells to the liposome-plasmid complexes, number of cell passages and cell types on GFP gene transfection of mouse somatic cells. The maximal GFP transgene expression (30.7%) was achieved when mouse fetal fibroblast cells (MFFC) at 70%-90% confluence of passage 3 were exposed for 6 h to the complexes of 4 microg liposome (LipofectAMINE) and 0.3 microg plasmid (pEGFP-N1). Under these conditions, we compared the effect of the number (from primary to 15) of passages on the transfection efficiency of MFFC. The transfection efficiency of MFFC was 10.0%, 28.9% and 7.2% at the primary, 3rd and 15th passage, respectively, which indicated that the transfection efficiency decreased with passaging. When MFFC, mouse oviductal epithelial cells (MOEC) and mouse granulosa cells (MGC) were transfected at passage 3, the transfection efficiency was 27.8%, 13.7% and 14.2%, respectively, under the described transfection conditions. When the cell cycle stages of different cell types at transfection were examined, it was found that 17.2% of MFFC, 8.7% of MOEC and 9.9% of MGC were at M phases of the cell cycle. Examination of the cell cycle stages of MFFC at different passages showed that MFFC at the third passage had the highest percentage of M cells and the percentage decreased afterwards. This suggested that the transfection efficiency was correlated with the percentages of cells at M phase, and provided essential data for improvement of the transfection efficiency.     

五株乳酸菌和三株芽孢杆菌的生物学特性和功能

【背景】乳酸菌和芽孢杆菌是应用于生产最多的益生菌,但不同菌株间的生长特性均不相同,因此了解菌株的生物学特性具有重要意义。【目的】研究菌株的生物学特性,能合理地开发和利用菌株,以保证菌株生产应用的安全性。【方法】活化后鉴定5株乳酸菌和3株芽孢杆菌并对其形态进行观察,探究菌株的生长曲线、产酸能力及最适生长条件,测定菌株的抑菌活性和产酶性能,同时探究菌株的益生性和安全性。【结果】五株乳酸菌分别编号鉴定为干酪乳杆菌R1、副干酪乳杆菌R2、香肠乳杆菌R3、福莱乳杆菌R4和唾液乳杆菌R5;3株芽孢杆菌分别编号命名为贝莱斯芽孢杆菌Y1、枯草芽孢杆菌Y2和地衣芽孢杆菌Y3。八株菌形态结构均不相同但都为杆状,均在2–10 h为对数生长期,18–24 h为稳定期,培养24 h时乳酸菌和芽孢杆菌的活菌数均保持在10⁹和10⁸ CFU/mL,最适生长温度为37.0℃。乳酸菌具有较强的产酸能力和抑菌活性,芽孢杆菌有较强的产酶性,在人工胃液中都有较强的耐受性。八株菌都无溶血活性、无毒力基因、对抗生素都保持中度敏感以上;其中唾液乳杆菌有四环素耐药基因,但对四环素抗性为中度敏感。【结论】八株菌生长繁殖速度快,乳酸菌产酸能力和抑菌活性较强,芽孢杆菌具有较强的产酶性能,在体外具有较好的益生性和安全性,可应用于生产实践。     

水培条件下营养元素对枳幼苗根毛发育及根生长的影响

以柑橘砧木枳实生苗为试材,研究水培条件下N、P、 K、Ca、Mg、Fe和Mn等7种营养元素分别缺乏对其根系主根长度、侧根数和主、侧根根毛密度、根毛长度及根毛直径等的影响．结果表明: 水培条件下,不同缺素处理枳实生幼苗的根毛均能生长,但根毛主要集中在近根基段,根尖处分布较少;侧根的根毛密度显著大于主根,而其根毛长度显著小于主根.不同缺素处理对根毛的生长发育影响较大,主根根毛密度为55.0~174.3 条·mm^-2．与对照相比,缺Ca诱发主根的根毛密度、长度显著增加;缺P使主根的根基段、中段及侧根的根毛密度、长度显著增加;缺Fe使主根根尖段根毛密度显著增加,而长度显著降低;缺K使主根、侧根的根毛密度、长度及根毛直径均显著降低;缺Mg使主根根毛长度显著增加．各处理主根的生长较一致;侧根除缺N、Mg处理外,其他处理均出现脱落后再生的现象.