昆虫产卵分泌物的产生方式、成分及功能

产卵是昆虫生活史中的重要阶段。绝大多数昆虫产卵前后都会产生分泌物用于卵的粘着和覆盖, 这些分泌物起到保护卵及初孵幼虫的作用;有些昆虫还通过产卵分泌物对卵进行标记, 进而影响种群、个体间对现有资源的有效利用。产卵分泌物是昆虫进化过程中的一种适应, 一般具有特定的生理和生态学功能, 影响着种群数量的变化。本文阐述了不同昆虫产卵分泌物的产生方式及物理性状, 并对产卵分泌物的组成、功能、分泌器官以及产卵分泌物中微生物的研究进展进行了综述。     

Characterization of antimicrobial peptides isolated from the skin of the Chinese frog, Rana dybowskii

The skins of amphibians secrete small antimicrobial peptides that fight infection and are being explored as potential alternatives to conventional antibiotics. In this study we combined mass spectrometry with cDNA sequencing to examine antimicrobial peptides in skin secretions from the Chinese frog Rana dybowskii. Thirteen peptides having precursor sequences that resemble known antimicrobial peptides from this genus were identified, ten of which were members of previously described peptide families based on their primary structures; i.e., brevinin-1, Japonicin-1, brevinin-2 and temporin. The other three peptides from R. dybowskii, which were named dybowskin-1CDYa, dybowskin-2 CDYa and dybowskin-2CDYb, had different amino acid compositions and little sequence similarity to known antimicrobial peptides. The carboxyl terminus of dybowskin-1CDY lacked amidation and is therefore clearly distinct from temporin peptides, whereas dybowskin-2CDYa and dybowskin-2CDYb consisted of 18 amino acids and were rich in Arg residues. Chemically synthesized peptides corresponding to mature dybowskin-1CDYa and dybowskin-2CDYa had strong antimicrobial activity and caused little hemolysis of human erythrocytes, suggesting they may serve as interesting templates for the development of novel antibiotics.     

Fatty acid composition of cholesteryl esters of human meibomian gland secretions

Very long chain cholesteryl esters (CE) are a major group of lipids found in meibomian gland secretions (MGS, also called meibum). MGS are produced by the meibomian glands of human and animal eyelids. They are a critical part of the tear film which covers the exposed ocular surface and serves various physiological roles. The composition of CE of MGS is complex, and still remains poorly understood. Here, a liquid chromatography-ion trap mass spectrometry (LC-MS) procedure developed to analyze CE is described, and a detailed composition of human meibomian CE is reported.MGS were collected from donors, analyzed without any modifications by LC-MS in positive and negative ion modes (PIM and NIM), and quantified using lipid standards where available.CE comprised about 30% of human meibum by mass. More than 40 individual CE species were found and characterized. In PIM, CE were observed as spontaneously in-source generated product ions m/z 369. The signals of the proton adducts of intact CE (M+H)⁺ were of very low intensity. In NIM, all tested CE spontaneously fragmented in-source producing signals of their respective FA. By combining the LC and MS information, the most abundant CE were found to be based on FA ranging from C₁₆ to at least C₃₂ in the following order C_26:0 > C_25:0 > C_24:0 > C_27:0 > C_24:1 = C_18:1 = C_20:0 > other CE.We conclude that the FA composition of CE can be successfully established in LC-MS experiments conducted in NIM. Meibomian CE have a large presence of both saturated and unsaturated FA with an average molar ratio of 4 to 1, respectively.     

免疫细胞分泌研究进展

细胞分泌活动涉及一系列复杂的分子活动和信号转导过程,它是许多重要生命活动如神经递质的释放、内分泌激素分泌、蛋白质转运等的基础。故而有关细胞分泌活动的研究近年在国际上形成一个新的热点。有关免疫细胞中细胞因子的分泌机制研究的较少,将膜电容测量、电化学测量、光学测量、胞内信号分子光解等近年发展起来的生物物理方法引入免疫学领域,对研究免疫细胞的分泌调控和信号转导意义重大。     

六种常见两栖类动物皮肤分泌物的生物活性比较

对6种常见两栖类动物皮肤水溶性分泌物的生物学尖性与酶学活性进行了研究,其中,红瘰疣螈（Tylototriton verrucosus),大蹼铃蟾（Bombina maxima),华西蟾蜍（Bufo andrewsi)皮肤分泌物对小白鼠具有致死毒性,对小白鼠腹腔注射的半数致死剂量分别为11．5,18．8和264mg/kg。而沼蛙（Rana nigromaculata),泽蛙（Rana guentheri),黑斑蛙（Rana limnocharis)的皮肤分泌物在小白鼠腹腔注射剂量达到500mg/kg时,仍不显示致死毒性。红瘰疣螈,大蹼铃蟾皮肤分泌物的毒性成分为3－60kDa的多态与蛋白质。6种两栖类动物皮肤分泌物都具有蛋白酶水解活性与胰蛋白酶抑制活性。红瘰疣螈,大蹼铃蟾和华西蟾蜍皮肤分泌物具有广谱抗菌活性,大蹼铃蟾,华西蟾蜍皮肤分泌的具有肿瘤细胞细胞毒活性。6种皮肤分泌物对哺乳类血液凝固系统无显著影响。仅发现红瘰疣螈皮肤分泌物具有磷脂酶A2活性。6种皮肤分泌的均无乙酰胆碱酯酶活性。     

利用回交法与Wx基因分子标记辅助选择培育糯性小麦

以中国春糯性位点全套近等基因系为研究材料,对小麦Wx基因的6个STS标记和1个CAPS标记进行了筛选。改良PCR扩增条件以及产物检测方式后,从这些标记中筛选出3个标记,包括鉴定Wx-A1、Wx-D1位点的2个共显性STS标记和Wx-B1位点的1个显性STS标记,用于本研究中糯性小麦的分子标记辅助育种。在育种过程中,首先配制全糯材料“98Y1441”与推广品种“川育12”的杂交组合,采用籽粒碘染法从其F2种子中选择全糯基因型个体与回交亲本川育12杂交,如此反复自交、回交,历经数代异地加代繁殖得到BC5F2代回交改良群体。利用上述3个分子标记从该群体中筛选出了8种Wx基因型,经卡方检验,其分离比符合3对基因的分离比例,其中基因型为aabbdd的植株有2株,直链淀粉含量分别为1.81%和0.82%,为全糯小麦;基因型为AAbbdd, aabbDD的部分糯性植株各有1株,直链淀粉含量分别为15.24%和17.57%。研究中获得的BC5 F2代群体的农艺性状接近回交亲本,并明显优于全糯材料“98Y1441”,表明采用回交法与Wx基因分子标记辅助选择相结合,有助于培育高产、优质的全糯和部分糯小麦。     

Multi-step Preparation Technique to Recover Multiple Metabolite Compound Classes for In-depth and Informative Metabolomic Analysis

Metabolomics is an emerging field which enables profiling of samples from living organisms in order to obtain insight into biological processes. A vital aspect of metabolomics is sample preparation whereby inconsistent techniques generate unreliable results. This technique encompasses protein precipitation, liquid-liquid extraction, and solid-phase extraction as a means of fractionating metabolites into four distinct classes. Improved enrichment of low abundance molecules with a resulting increase in sensitivity is obtained, and ultimately results in more confident identification of molecules. This technique has been applied to plasma, bronchoalveolar lavage fluid, and cerebrospinal fluid samples with volumes as low as 50 µl.  Samples can be used for multiple downstream applications; for example, the pellet resulting from protein precipitation can be stored for later analysis. The supernatant from that step undergoes liquid-liquid extraction using water and strong organic solvent to separate the hydrophilic and hydrophobic compounds. Once fractionated, the hydrophilic layer can be processed for later analysis or discarded if not needed. The hydrophobic fraction is further treated with a series of solvents during three solid-phase extraction steps to separate it into fatty acids, neutral lipids, and phospholipids. This allows the technician the flexibility to choose which class of compounds is preferred for analysis. It also aids in more reliable metabolite identification since some knowledge of chemical class exists.     

Impact of low oxygen on the secretome of human adipose-derived stromal/stem cell primary cultures

Tissue fibrosis can lead to organ dysfunction, patient morbidity, and mortality. Adipose-derived Stromal/stem Cells (ASCs) represent a potential therapeutic. Immediately following grafting, ASCs would reside in a lower O₂ environment. ASC secretome was examined under 5% O₂ (“low O₂”) and 21% O₂ (“ambient O₂”) culture conditions. ASCs from five female donors were cultured in low or ambient O₂ conditions for 3 days and pooled conditioned medium was compared by two-dimensional liquid chromatography and tandem mass spectrometry (2D-LC–MS/MS). Of 71 proteins identified, five proteins involved in extracellular matrix (ECM) remodeling exhibited ≥2-fold decrease under low O₂ culture and were confirmed by Western immunoblot and qRT-PCR: fibronectin 1, TGF-β1-induced protein (βig-h3), osteonectin, and collagens type 1α₁ and α₂. ELISAs performed using 10 donors also confirmed significant decreases during low O₂ culture in 4–6 ASC donors. For low abundant proteins, a 36 cytokine/chemokine array was performed. Fifteen cytokines/chemokines including Type 2 cytokines IL-13, MCP-1, and CD40 ligand were detected in ambient O₂ ASC medium. IL-6 was detected in low O₂ but not ambient O₂ ASC medium. These findings demonstrate that low O₂ ASC exposure resulted in reduced ECM protein and Type 2 cytokine secretions that are significant with regard to inflammation in fibrosis.     

蜡梅花被片表层形态观察及其意义

为探究蜡梅花被片表层蜡质的微形态结构特征和差异性,采用扫描电子显微镜对蜡梅和山蜡梅的花被片进行观察。结果表明:蜡梅花被片表层无明显蜡质覆盖物,细胞排列平滑,内表层有加厚透明状覆盖物; 山蜡梅花被片表层有厚蜡质覆盖物和表皮毛; 山蜡梅、蜡梅花被片均无气孔。以上独特的结构形态对于蜡梅花开放于寒冷季节,应对外界环境胁迫可能有一定的保护作用和生态意义。