间断低氧对大鼠下丘脑超微结构及前增食欲素水平的影响

目的探讨睡眠中间断低氧对大鼠下丘脑前增食欲素及受体水平的影响以及下丘脑超微结构的变化。方法大鼠分成对照组、间断低氧组和持续低氧组,分别给予吸入空气,持续低氧和间断低氧气体,并在实验开始后1d、3d、1w和4w应用RT-PCR方法测定大鼠下丘脑前增食欲素及受体水平,分析其间的变化关系,电镜观察下丘脑的超微结构变化。结果与对照组和持续低氧组比较,间断低氧4w后大鼠下丘脑前增食欲素mRNA水平明显降低,受体水平升高,但在持续低氧和对照组之间无明显差异。在低氧后1d、3d、7d后大鼠下丘脑前增食欲素mRNA降低,受体水平升高,在4w后,持续低氧组则接近正常。急性持续低氧大鼠超微结构变化更严重,而慢性间断低氧变化更持久。结论慢性间断低氧可以引起下丘脑前增食欲素下降及受体水平升高,急性持续低氧也可引起上述变化,而慢性持续低氧未引起增食欲素改变;慢性间断低氧大鼠下丘脑超微结构表现为严重而持久的变化。     

武汉桂子山地区冬季鸟类群落结构及多样性研究

2005年11月～2006年3月,对武汉桂子山地区冬季鸟类群落进行了调查,共记录鸟类44种,隶属10目22科。有国家Ⅱ级保护鸟类2种,湖北省重点保护鸟类16种;留鸟32种,冬候鸟11种,旅鸟1种,留鸟构成了冬季鸟类的主体;该地区冬季鸟类密度为4．30只／hm^2,物种多样性指数为2．6477,均匀性指数为0．6997。居民点、乔灌林、绿地和水域生境中,乔灌林鸟类多样性最高,数量最多。     

A novel RGD-toxin protein,Lj-RGD3, from the buccal gland secretion of Lampetra japonica impacts diverse biological activities

RGD (Arg-Gly-Asp) motif toxin proteins from snake venoms, saliva glands secretion of leech or tick have typical characteristics of inhibiting platelet aggregation, angiogenesis, and tumor growth. Here we report cloning and characterization of a novel RGD-toxin protein from the buccal gland of Lampetra japonica. In an attempt to study the activities of anticoagulant in the buccal gland secretion of L. japonica, we established buccal gland cDNA library and identified a gene encoding a predicted protein of 118 amino acids with 3 RGD motifs. The predicted protein was named Lj-RGD3. We generated the cDNA of Lj-RGD3 and obtained the recombinant protein rLj-RGD3. The polyclonal antibodies against rLj-RGD3 recognized the native Lj-RGD3 protein in buccal gland secretion in Western blot analyses. The biological function studies reveal that rLj-RGD3 inhibited human platelet aggregation in a dose-dependent manner with IC₅₀ value at 5.277 μM. In addition, rLj-RGD3 repressed bFGF-induced angiogenesis in the chick chorioallantoic membrane model. rLj-RGD3 also inhibited the adhesion of ECV304 cells to vitronectin. Furthermore, rLj-RGD3 induced apoptosis and significantly inhibited proliferation, migration, and invasion evoked by bFGF in ECV304 cells. Taken together, these results suggested that rLj-RGD3 is a novel RGD-toxin protein possessing typical functions of the RGD-toxin protein.     

中枢神经细胞的高尔基复合体ACP活性分布的电镜观察

本文采用电镜金属盐法—酸性磷酸酶(ACP)细胞化学技术,用30mmol/L pipes缓冲液配制低浓度戊二醛进行固定。对成年大鼠的大脑大锥体细胞,小脑浦肯野氏细胞,脊髓前角运动细胞的高尔基复合体的ACP活性进行了实验研究和探讨。结果发现ACP活性分布在高尔基复合体的部份转移泡、浓缩泡及GERL部位。高尔基复合体呈ACP阳性反应,并显示出多种形态。     

小肠平滑肌胰岛素受体的特征及其蛋白激酶活性

 本实验对狗小肠平滑肌中胰岛素受体的结构和特征进行了分析研究。通过麦胚凝集素琼脂糖和两次Sepharose-CL-6B凝胶层析从平滑肌中纯化胰岛素受体,达到电泳纯。SDS-聚丙烯酰胺凝胶电泳证明胰岛素受体是由两个亚基组成的,分子量分别为135kD和90kD。磷酸化实验证明平滑肌胰岛素受体具有胰岛素依赖性蛋白激酶活性,能催化自身的β亚基磷酸化和底物的磷酸化。Scatchard分析表明胰岛素和受体的结合呈(?)协同效应,最大结合率为13μg胰岛素/mg蛋白质。     

Species composition and diversity of soil animals in the oil shale dump in Maoming, Guangdong Province, China

Pollution from the oil shale dump in Maoming, Guangdong Province, China, was a major social problem due to the great amount of environmental damage it caused. Therefore, a great deal of attention needed to be paid for the ecological restoration and reconstruction. The objective of this study was to investigate the species composition of soil fauna and its diversity in oil shale dumps after the application of different ecological restoration schemes in order to understand the impact they had on ecological restoration. Three plots were selected on an oil shale dump near the city of Maoming. The “north plot” was a newly-planted young forest mixed with various tree species, while the “south plot” was a 20-year-old Acacia auriculaeformis forest, and the “control plot” was a 20-year-old naturally-recovering grassland. Soil animals, mainly including macro-meso groups, were collected by hand-sorting and Tullgren funnel extraction. They were then identified to family or genus level with only a small portion to order (e.g. Chllopoda) or species (e.g. Isopoda) level. The specimens obtained in the present study was 11164 individuals, belonging to 27 orders and 110 families or genera. The Shannon index (H′), DG_s (based on species) and DG_g (based on groups) were used to analyze the diversity of soil animals between different plots. The major results were as follows: A total of 33 families or genera belonging to nine orders were found in the “north plot”. The main group was Caritermes, accounting for 63.4% of the total, followed by Tetramorium with 21.3%. Hymenoptera, mainly Formicidae, had more genera than others, accounting for 80% of the total genera in this group. The diversity of soil animals in this plot was very low because the H′ index was only 1.2, while the DG_s index was 4.0 and the DG_g index was 1.3. A total of 61 families or genera belonging to 23 orders were found in the “south plot”. Malmcoangelia and Tetramorium were the main groups, which accounted for 60.3% and 10.2%, respectively. Two genera of Annelda and two genera of Isopoda only accounted for 2.6% and 1.9%, respectively, but they were considered to be major groups due to their large body sizes and the distinct characteristics of their habitat. Acarina had a greater number of individuals and families or genera with its individual number accounting for 67.5% of the total, and the number of families or genera of this group account for 70% in this plot. The diversity indexes (H′, DG_s and DG_g) in this plot were significantly higher than those in the “north plot”, and were 1.65, 16.7 and 7.75, respectively. In the “control plot”, there were 67 families or genera of soil animals belonging to 23 orders. The main groups were Tetramorium (20%), Lasius (17.1%), Bothriomymex of Formicidae (13.8%), and Malmcoangelia of oribatid mites (14.5%). Formicidae of Hymenoptera was the group with the maximum number of individuals, accounting for 51.0%, while Diplopoda had the most families or genera. The H′ and DG_s indexes, being 2.54 and 17.7, were higher than those in the “south plot”, while the DG_g index of 7.20 was lower than that in the “south plot”. The results showed that the species composition and diversity indexes were higher in the “south plot” than in the “north plot” and the “control plot”, which demonstrated that using Acacia auriculaeformis forest to restore the oil shale dump was an effective approach in terms of soil biodiversity.     

Efficient flow cytometric assay for platelet-leukocyte aggregates in whole blood using fluorescence signal triggering.

BACKGROUND: Platelet-leukocyte aggregates (PLAs) may be important in thrombotic and inflammatory disease states, but accurate assessment of PLA formation in vivo is hampered by the propensity for in vitro artefacts caused by sample manipulation. A whole blood flow cytometric assay for circulating PLAs, based on minimal sample manipulation, was thus developed. METHODS: Citrated whole blood was labeled with a RPE-CD45 MAb (leukocyte marker) and an FITC-CD42a (GPIX) MAb (platelet marker). The latter was used to avoid possible influences of platelet glycoprotein proteolysis by neutrophil-derived proteases. The samples were mildly fixed with 0.5% formaldehyde saline. The cytometer was triggered by RPE-CD45 fluorescence. Leukocyte subpopulations were separated according to their typical light scattering and CD45 expression. RESULTS: Minimal sample manipulation and mild sample fixation resulted in minor in vitro artefacts and good sample stability. Fluorescence triggering increased the efficiency of the flow cytometric analysis approximately 5-fold compared with triggering with light scatter, and allowed discrimination of leukocyte subpopulations. The majority of PLAs involved monocytes and neutrophils, rather than lymphocytes, both without and with in vitro stimulation by ADP or thrombin. A cocktail of blocking MAbs to CD62P, CD15, GPIIb/IIIa and the CD11b/CD18 complex had no effect on unstimulated samples, whilst totally inhibiting aggregation induced by 10(-5) M ADP, suggesting that the PLAs in unstimulated blood were preformed in vivo. CONCLUSIONS: This whole blood flow cytometric assay for PLAs is simple and efficient, and appears to reflect closely platelet-leukocyte aggregates in circulating blood in vivo.