鸭C4BPα的克隆、表达及其与鸭疫里默氏菌的互作

为研究鸭C4结合蛋白(C4b-binding protein,C4BP)与鸭疫里默氏菌(Riemerella anatipestifer,RA)的相互作用,对鸭C4BPα进行克隆、原核表达,免疫小鼠制备多克隆抗体,并利用间接免疫荧光试验及斑点杂交试验验证C4BP与RA的相互作用。结果显示,鸭C4BPα核苷酸序列全长为1230bp,与鸡C4BPα的相似性最高(82.1%);系统进化树分析发现,鸭C4BPα与鸡C4BPα处于同一系统进化树分支上,两者遗传进化关系最近;C4BPα在大肠杆菌Escherichia coli BL21 (DE3)中能高效表达,重组蛋白以胞内可溶性形式存在;多克隆抗体效价超过1∶10000,并且可以与重组蛋白发生特异性反应;间接免疫荧光试验和斑点杂交试验结果显示RA与鸭C4BP可以发生相互作用。研究结果为进一步揭示RA的致病机制奠定了基础。     

Effets des différentes fractions du gradient de Percoll discontinu sur la mobilité des spermatozoides humains

The centrifugation on discontinued Percoll gradient is a technique of sperm preparation, which has enabled better success of in-vitro fertilization. In this work, we have analysed the movements of spermatozoa separated in different Percoll fractions after 4 hours of incubation at 37°C and under 5% of CO₂. These measures are made by computer assisted videomicrography (Cell Soft). From this study, we notice that the more the Percoll concentration increases, the greater is the curvilinear velocity, the linearity and the lateral amplitude of the head. The 100% and 90% Percoll fractions help to get spermatozoa with good movement features, indicating the use of this technique in medically assisted procreation.     

A 63-kDa protein with androgen-binding activity is not from the androgen receptor.

We have found a new protein in the heart of rat and mice that can be selectively and covalently labelled with the synthetic androgen analog mibolerone. Binding is specific as it can be displaced by excess radioinert ligand. The protein is prominently expressed in liver, kidney, and heart, but not in skeletal muscle. It is water soluble and found in the cytosol. Under denaturing conditions it has a molecular weight of 63,000 and appears on two-dimensional gels with an isoelectric point of 6.3. The protein's affinity for androgen is lower than that of the androgen receptor and it is about 100-fold more abundant than the receptor in the heart. Expression of the protein is not induced by androgen. The presence of this protein in testicular feminization (tfm) mice with a genetical defect of the androgen receptor rules out that it is the androgen receptor or a portion thereof. The biological role of this protein is not yet known.     

Declined miR‐181a‐5p expression is associated with impaired natural killer cell development and function with aging

MicroRNAs (miRNAs) regulate gene expression and thereby influence cell development and function. Numerous studies have shown the significant roles of miRNAs in regulating immune cells including natural killer (NK) cells. However, little is known about the role of miRNAs in NK cells with aging. We previously demonstrated that the aged C57BL/6 mice have significantly decreased proportion of mature (CD27⁻CD11b⁺) NK cells compared with young mice, indicating impaired maturation of NK cells with aging. Here, we performed deep sequencing of CD27⁺ NK cells from young and aged mice. Profiling of the miRNome (global miRNA expression levels) revealed that 49 miRNAs displayed a twofold or greater difference in expression between young and aged NK cells. Among these, 30 miRNAs were upregulated and 19 miRNAs were downregulated in the aged NK cells. We found that the expression level of miR‐l8la‐5p was increased with the maturation of NK cells, and significantly decreased in NK cells from the aged mice. Knockdown of miR‐181a‐5p inhibited NK cell development in vitro and in vivo. Furthermore, miR‐181a‐5p is highly conserved in mice and human. MiR‐181a‐5p promoted the production of IFN‐γ and cytotoxicity in stimulated NK cells from both mice and human. Importantly, miR‐181a‐5p level markedly decreased in NK cells from PBMC of elderly people. Thus, our results demonstrated that the miRNAs profiles in NK cells change with aging, the decreased level of miR‐181a‐5p contributes to the defective NK cell development and function with aging. This opens new strategies to preserve or restore NK cell function in the elderly.     

Diterpenoids from the Whole Plants of Ajuga nipponensis and Their Inhibition of RANKL-Induced Osteoclastogenesis

Two new diterpenoids, ajudecunoid A ( 1 ) and ajudecunoid B ( 14 ), along with thirteen known diterpenoids, were isolated from the whole plants of Ajuga nipponensis Makino. Their structures were elucidated by the extensive spectroscopic analysis (UV, IR, MS, and NMR). The absolute configurations of ajudecunoid A ( 1 ) and ajudecunoid B ( 14 ) were defined through analysis of X-ray crystallography. Fifteen compounds were evaluated for inhibition of the formation of osteoclasts in bone marrow-derived macrophages (BMM) cells. Two neo-clerodane diterpenoids ajuganipponin B ( 5 ) and (12S)-6α,19-diacetoxy-18-chloro-4α-hydroxy-12-tigloyloxy-neo-clerod-13-en-15,16-olide ( 12 ) showed significant inhibition of osteoclastogenesis with IC₅₀ values of 0.88 and 0.79 μM, respectively. Here we firstly reported diterpenoids with anti-osteoclastogenesis activity from A. nipponensis.