鲫幼鱼耳石荧光标记效果

为确认茜素络合物对鲫（Carassius auratus）耳石进行有效标记的可行性,以便为鲫甚至其他鲤科鱼类标志放流技术的开发及效果评估提供一定的借鉴,本研究以孵出后90 d的鲫幼鱼为研究对象,设置单一浓度（100 mg/L）的茜素络合物浸泡标记5 d,分析茜素络合物在耳石上的沉积情况以及不同耳石在不同后续饲养天数的动态变化。结果表明,矢耳石、微耳石和星耳石上在可见光、绿色和蓝色激发光下都检测到了良好的茜素络合物标记环,标记率和存活率均为100%。但不同耳石的茜素络合物标记效果不同,荧光下,星耳石的标记效果最显著,微耳石次之;可见光下,微耳石的标记效果最好,星耳石次之。随着后续饲养天数的延长,可见光下标记逐渐减弱,至20 d时基本消失,而在绿色和蓝色激光下标记环荧光强度无减弱迹象,能长久保持,且在蓝色激发光下标记环更易被观测到。上述结果结合鲫生长、存活和行为正常等情况综合显示,在100 mg/L茜素络合物溶液中浸泡标记鲫幼鱼5 d,其耳石可以获得满意的标记效果。     

Variable expression of lineage regulators in differentiated stromal cells indicates distinct mechanisms of differentiation towards common cell fate

Mesenchymal stem cells (MSCs) possess a multi-lineage differentiation capacity that makes them important players in the field of regenerative medicine. MSC populations derived from different tissues or donors have been shown to exhibit variable gene expression patterns. Further, it is widely acknowledged that MSC isolates are heterogeneous mixtures of cells at different developmental stages. However, the heterogeneity of expression of lineage regulators has not been linked to differentiation potential of different MSC populations towards mesenchymal lineages. Here, we analyzed variation of expression of differentiation markers across whole population and between single differentiating cells of multipotent stromal cell populations derived from adipose tissue (AdMSCs) and skin (FBs) of seven donors. The results of the analyses show that all cell populations exhibit similar differentiation potential towards adipocyte, osteoblast and chondrocyte lineages despite tissue type- and donor-specific variations of expression of differentiation-associated genes. Further, we detected variable expression of lineage regulators in individual differentiating cells. Together, our data indicate that single cells of stromal cell populations could use distinct molecular mechanisms to reach a common cell fate.     

BSA–boronic acid conjugate as lectin mimetics

We report bovine serum albumin (BSA)–boronic acid (BA) conjugates as lectin mimetics and their glyco-capturing capacity. The BSA–BA conjugates were synthesized by amidation of carboxylic acid groups in BSA with aminophenyl boronic acid in the presence of EDC, and were characterized by Alizarin Red S (ARS) assay and SDS–PAGE gel. The BSA–BA conjugates were immobilized onto maleimide-functionalized silica beads and their sugar capturing capacity and specificity were confirmed by ARS displacement assay. Further, surface plasmon resonance (SPR) analysis of the glyco-capturing activity of the BSA–BA conjugates was conducted by immobilizing BSA–BA onto SPR gold chip. Overall, we demonstrated a BSA–BA-based lectin mimetics for glyco-capturing applications. These lectin mimetics are expected to provide an important tool for glycomics and biosensor research and applications.     

用荧光物质浸泡标记胭脂鱼仔、稚鱼耳石

用茜素络合物和盐酸四环素溶液分别对胭脂鱼（Myxocyprinus asiaticus）仔、稚鱼浸泡标记,结果表明,100～200mg／L的茜素络合物溶液浸泡24h对胭脂鱼仔、稚鱼耳石有很好的标记效果。相同浸泡浓度下,随着日龄的增长,耳石上的荧光反应强度降低。三对耳石中,微耳石和矢耳石对茜素络合物较敏感,星耳石敏感性则较低。盐酸四环素溶液对仔、稚鱼耳石的标记效果很差,浸泡液浓度为100mg／L和120mg/L时,仅能在微耳石上检测到较弱的荧光标记,而150mg／L及以上浓度的浸泡液对稚鱼有较高的致死作用。因此,茜素络合物是对胭脂鱼早期鱼苗进行化学标记比较合适的荧光物质,而盐酸四环素不适于标记该鱼的耳石。在对标记鱼进行荧光检测时,矢耳石和微耳石是适合的材料。     

茜素红S对中华倒刺鲃幼鱼不同组织抗氧化酶活性和脂质过氧化的影响

以中华倒刺鲃幼鱼[平均体长（12.3±0.6）cm,平均体重（41.8±3.6）g]为研究对象,通过测定不同浓度茜素红S溶液（ARS）浸泡24h后肝脏、脑和鳃组织中抗氧化酶（SOD、CAT、GSH-Px）的活性和丙二醛（MDA）含量,评估ARS对中华倒刺鲃幼鱼的生理生化影响。结果表明:除脑组织GSH-Px随着ARS浓度的升高而不断升高外,肝脏、鳃和脑组织的抗氧化酶活性均表现为低浓度被诱导而高浓度受抑制的规律,与ARS浓度呈抛物线型剂量效应关系。肝脏、鳃、脑组织抗氧化酶活性达到最大值所对应的浓度分别是300、300和400 mg/L。除鳃部MDA含量随着茜素红S浓度升高不断升高外,脑和肝脏组织的丙二醛含量随着ARS浓度的升高呈现出先降低后上升趋势。根据试验的结果,推荐120-200 mg/L为ARS染色标记中华倒刺鲃幼鱼（体长10 cm）的适宜浓度。     

An alternative Alcian Blue dye variant for the evaluation of fetal cartilage

BACKGROUND: The most comprehensive evaluation of vertebrate skeletal development involves the use of Alizarin Red S dye to stain ossified bone and various other dyes to stain cartilage. The dye used most widely to stain fetal cartilage in rodents and rabbits is Alcian Blue 8GX. However, the global supply of this specific dye has been exhausted. Several forms of the dye marketed as Alcian Blue 8GX are now available, although they are not synthesized via the original 8GX manufacturing process. METHODS: One new Alcian Blue 8GX form and two Alcian Blue dye variants were evaluated in rats and rabbits using standard staining procedures. The staining quality of these dyes were evaluated relative to the original form of Alcian Blue 8GX based on cartilage uptake of the dye, clarity of the cartilaginous components, staining intensity of the dye, and overall readability of the specimens under stereomicroscopic evaluation. RESULTS: Staining with the newer form of Alcian Blue 8GX resulted in poor staining quality. The Alcian Blue-Pyridine variant performed well, although staining intensity was less than optimal. The Alcian Blue-Tetrakis variant provided staining characteristics that were most similar to the original form of Alcian Blue 8GX. CONCLUSIONS: Alcian Blue-Tetrakis was markedly better in its ability to stain fetal cartilage than the newer form of Alcian Blue 8GX.     

茜素络合物对唐鱼耳石标记效果以及生长和存活率的影响

用茜素络合物(ALC)对唐鱼(Tanichthys albonubes)进行荧光标记,通过测定唐鱼耳石的标记率、死亡率、SGR和DWG等指标,讨论不同浓度ALC对唐鱼耳石标记效果和对其生长和存活的影响,探讨应用该标记技术追踪唐鱼野外种群迁移行为的可行性。结果表明:在温度28~30℃,浓度50 mg·L-1、80 mg·L-1茜素络合物溶液中浸泡24 h,唐鱼仔鱼、稚鱼死亡率为0,其中80 mg·L-1浓度处理组耳石荧光标记环检出率为100%,而在100mg·L-1浓度下,仔、稚鱼死亡率均达到80%;幼鱼在浓度为80、100 mg·L-1条件下无死亡,其中100 mg·L-1浓度处理组耳石标记环检出率为100%,而150 mg·L-1浓度下幼鱼死亡率为44%;成鱼在100、150 mg·L-1浓度下无死亡,且耳石标记率均为100%,而在200mg·L-1浓度下成鱼死亡率达到100%;此外,稚鱼在80 mg·L-1最适浓度下浸泡24 h后继续饲养90 d,标记组和对照组死亡率均为0,两组体质量、SGR和DWG差异不显著;唐鱼仔稚鱼、幼鱼和成鱼耳石标记的最适茜素络合物浓度分别为80、100和150 mg·L-1。     

SPATIAL AND TEMPORAL VARIABILITY IN BIOMASS AND PRODUCTION OF PSAMMOPHYTIC HALIMEDA INCRASSATA (BRYOPSIDALES,CHLOROPHYTA) IN A CARIBBEAN REEF LAGOON1

Estimates for the production of calcium carbonate by Halimeda spp. have been based on limited measures in small areas or over short periods, subsequently extrapolated over larger temporal and spatial scales. The accuracy of these extrapolations depends on the variations in time and space of the parameters used for the derivations of the production, which were evaluated in the present study for Halimeda incrassata (J. Ellis) J. V. Lamour. in the Puerto Morelos reef lagoon, Mexican Caribbean. Growth, biomass, and CaCO₃ content of the branches of the thalli were determined at 1–3 monthly intervals from November 1997 until June 1999, using the marking technique with the coloring agent Alizarin Red‐S. Biomass exhibited great variability (V=25.4%), and fluctuations were closely related to changes in thallus density. Growth and CaCO₃‐content changes were related to the solar cycle, having coefficients of 15.4% and 2.5%, respectively. Additionally, calcified biomasses of the thalli were determined over a large spatial (31 stations in ～7 km² area) and time (6 y) scale. Spatial variability in calcified biomass was 59%, and the coefficient of variation attained its highest value (69%) for the samples collected over a 6 y period, from 1990 to 1996 (except 1995). Based on overall average values, branches of H. incrassata in the study area had a mean turnover of 30 d, with an annual production of 815 g CaCO₃ per square meter.     

Uncoupling of osteoblast–osteoclast regulation in a chemical murine model of Gaucher disease

Gaucher disease (GD) is caused by mutations in the GBA gene that confer a deficient level of activity of glucocerebrosidase (GCase). This deficiency leads to accumulation of the glycolipid glucocerebroside in the lysosomes of cells of monocyte/macrophage system. Type I GD is the mildest form and is characterized by the absence of neuronopathic affection. Bone compromise in Gaucher disease patients is the most disabling aspect of the disease. However, pathophysiological aspects of skeletal alterations are still poorly understood.