Mutual dependence of VIP/PACAP and CCK receptor signaling for a physiological role in duck exocrine pancreatic secretion

Unlike in rodents, CCK has not been established as a physiological regulator in avian exocrine pancreatic secretion. In the isolated duck pancreatic acini, 1 nM CCK was required for stimulation of amylase secretion, maximal effect being achieved at 10 nM; picomolar CCK was without effect. Vasoactive intestinal peptide (VIP)/pituitary adenylate cyclase activating peptide (PACAP) receptor (VPAC) agonists PACAP-38 and PACAP-27 (10(-12)-10(-7) M) alone had no effect, but made picomolar CCK effective. VPAC agonist VIP 10(-10)-10(-7) M stimulated amylase secretion marginally, but made CCK 10(-12)-10(-10) M effective also. PACAP-27 and VIP both shifted the maximal CCK concentration from 10(-8) to 10(-9) M. This sensitizing effect was mimicked by forskolin. CCK dose dependently induced intracellular Ca2+ concentration ([Ca2+]i) oscillations. PACAP-38 (1 nM), PACAP-27 (1 nM), VIP (10 nM), or forskolin (10 microM) alone did not stimulate [Ca2+]i increase, neither did they modulate CCK (1 nM)-induced oscillations; but when they were added to cells simultaneously exposed to subthreshold CCK (10 pM), calcium spikes emerged. Amylase secretion induced by the simultaneous presence of 10 pM CCK and VPAC agonists was completely blocked by removing extracellular calcium, but the protein kinase C inhibitor staurosporine (1 microM) was without effect. CCK (10 nM)-induced secretion was inhibited by CCK1 receptor antagonist FK480 (1 microM). Gastrin from 10(-12) to 10(-6) M did not stimulate amylase secretion nor did it (100 nM) induce [Ca2+]i increase. The above data suggest that duck pancreatic acini possess both CCK1 and VPAC receptors; simultaneous activation of both is required for each to play a physiological role.     

抗树鼩CD3ε单克隆抗体的制备及生物学特性鉴定

制备鼠抗树鼩CD3ε单克隆抗体,并对其生物学特性进行鉴定。     

我国商业化SPF级小鼠病原体污染分析

目的按现行国标《实验动物微生物学检测方法》和《实验动物寄生虫学检测方法》对我国商品化的无特殊病原体（specific pathogen free,SPF）小鼠进行微生物状况检测,为生产高质量的实验动物提供依据。方法对五家主要实验动物生产单位生产的ICR、KM、C57BL/6、BALB/c及BALB/c-nu品系的SPF级小鼠进行随机抽检。检测抽检小鼠所携带的微生物和寄生虫情况。结果在抽检的SPF级小鼠中,病毒污染主要包括呼肠孤病毒Ⅲ型（Reo-3）、小鼠肺炎病毒（PVM）和多瘤病毒（POLY）;主要细菌污染为肺炎克雷伯杆菌、金黄色葡萄球菌和绿脓杆菌。这些微生物病原体中,除条件性致病菌绿脓杆菌外,其他病原体对小鼠本身和实验研究均具有一定的影响。     

达斡尔族成人的体格,体型及半个多世纪来的变化

对内蒙古自治区莫力达瓦族３５３名（男１８７,女１６６）２０－６０岁达斡尔族进行了活体观察与测量,总结了达斡尔族的体格特征和类型。并在城乡之间和与半个世纪以前同一地区达斡尔族的体质资料之间进行了比较。     

不同产地丹参药材HPLC指纹图谱分析及4种菲醌类成分含量的比较

对7个产地丹参(Salvia miltiorrhiza Bge.)药材进行了HPLC分析并确定了其指纹图谱的共有模式谱,在此基础上比较了7个产地丹参药材中4种菲醌类成分(二氢丹参酮Ⅰ、丹参酮Ⅰ、隐丹参酮和丹参酮ⅡA)的相对含量。结果表明:来源于不同产地的丹参药材的HPLC图谱均有21个峰,其中有8个共有峰;各共有峰的相对保留时间基本相同,RSD值均小于0.086%;但相对峰面积差异明显。以4号峰(二氢丹参酮Ⅰ)为参照峰,初步构建了丹参药材的HPLC指纹图谱的共有模式谱,7个产地丹参药材的HPLC指纹图谱与共有模式谱的相似性良好,相似度值为0.916～0.973。不同产地丹参药材中4种菲醌类成分的相对含量有明显差异,其中,河南产药材中4种菲醌类成分含量均最高,而河北产药材中均最低;此外,不同产地丹参药材中4种成分的组成比例也有明显差异。根据实验结果,建议将丹参药材的HPLC指纹图谱的共有模式谱作为丹参药材质量控制和真伪辨别的标准之一。     

Mycobacterial Cytochrome P450 125 (Cyp125) Catalyzes the Terminal Hydroxylation of C27 Steroids

Cyp125 (Rv3545c), a cytochrome P450, is encoded as part of the cholesterol degradation gene cluster conserved among members of the Mycobacterium tuberculosis complex. This enzyme has been implicated in mycobacterial pathogenesis, and a homologue initiates cholesterol catabolism in the soil actinomycete Rhodococcus jostii RHA1. In Mycobacterium bovis BCG, cyp125 was up-regulated 7.1-fold with growth on cholesterol. A cyp125 deletion mutant of BCG did not grow on cholesterol and accumulated 4-cholesten-3-one when incubated in the presence of cholesterol. Wild-type BCG grew on this metabolite. By contrast, a parallel cyp125 deletion mutation of M. tuberculosis H37Rv did not affect growth on cholesterol. Purified Cyp125 from M. tuberculosis, heterologously produced in R. jostii RHA1, bound cholesterol and 4-cholesten-3-one with apparent dissociation constants of 0.20 ± 0.02 μm and 0.27 ± 0.05 μm, respectively. When reconstituted with KshB, the cognate reductase of the ketosteroid 9α-hydroxylase, Cyp125 catalyzed the hydroxylation of these steroids. MS and NMR analyses revealed that hydroxylation occurred at carbon 26 of the steroid side chain, allowing unambiguous classification of Cyp125 as a steroid C26-hydroxylase. This study establishes the catalytic function of Cyp125 and, in identifying an important difference in the catabolic potential of M. bovis and M. tuberculosis, suggests that Cyp125 may have an additional function in pathogenesis.     

Isolation,Diversity, and Antimicrobial Activity of Rare Actinobacteria from Medicinal Plants of Tropical Rain Forests in Xishuangbanna,China

Endophytic actinobacteria are relatively unexplored as potential sources of novel species and novel natural products for medical and commercial exploitation. Xishuangbanna is recognized throughout the world for its diverse flora, especially the rain forest plants, many of which have indigenous pharmaceutical histories. However, little is known about the endophytic actinobacteria of this tropical area. In this work, we studied the diversity of actinobacteria isolated from medicinal plants collected from tropical rain forests in Xishuangbanna. By the use of different selective isolation media and methods, a total of 2,174 actinobacteria were isolated. Forty-six isolates were selected on the basis of their morphologies on different media and were further characterized by 16S rRNA gene sequencing. The results showed an unexpected level of diversity, with 32 different genera. To our knowledge, this is the first report describing the isolation of Saccharopolyspora, Dietzia, Blastococcus, Dactylosporangium, Promicromonospora, Oerskovia, Actinocorallia, and Jiangella species from endophytic environments. At least 19 isolates are considered novel taxa by our current research. In addition, all 46 isolates were tested for antimicrobial activity and were screened for the presence of genes encoding polyketide synthetases and nonribosomal peptide synthetases. The results confirm that the medicinal plants of Xishuangbanna represent an extremely rich reservoir for the isolation of a significant diversity of actinobacteria, including novel species, that are potential sources for the discovery of biologically active compounds.The class Actinobacteria accounts for a high proportion of soil microbial biomass and contains the most economically significant prokaryotes, producing more than half of the bioactive compounds in a literature survey (46), including antibiotics (6), immunosuppressive agents (55), antitumor agents (18), and enzymes (64). Actinobacteria belonging to the genus Streptomyces, in particular, are excellent producers. The emergence of drug resistance in many bacterial pathogens and the current increase in the number of fungal infections has caused a resurgence of interest in finding new reserves of biologically active compounds (63). As the search for novel natural products continues, it becomes apparent that the rate of discovery of new compounds from soil streptomycetes has decreased, whereas the rate of reisolation of known compounds has increased (28). Recently, evidence has accumulated that rare actinomycete species, which are often very difficult to isolate and cultivate, might represent a unique source of novel biologically active compounds (4). On the other hand, new microbial habitats need to be examined in the search for novel bioactive compounds. One biologically important but relatively overlooked niche is the inner tissues of higher plants. Early studies have demonstrated that some actinobacteria can form intimate associations with plants and colonize their inner tissues. Frankia species and Streptomyces scabies can penetrate their hosts and establish either pathogenic or endophytic associations (5, 24). The actinomycetes that reside in the tissues of living plants and do not visibly harm the plants are known as endophytic actinobacteria (37). These actinobacteria are relatively unstudied and are potential sources of novel natural products for exploitation in medicine, agriculture, and industry (73).Endophytic actinobacteria have attracted attention in recent years, with increasing reports of isolates from a range of plant types, including crop plants (cereals, such as wheat and rice, as well as potatoes, carrots, tomatoes, and citrus) (2, 16, 62, 71, 74, 80) and medicinal plants (75, 88). The culturable endophytic actinobacteria from these plants were found to fall within a narrow species distribution: Streptomyces spp. were the predominant species, and Microbispora, Micromonospora, Nocardioides, Nocardia, and Streptosporangium were the common genera. Endophytic actinobacteria have been demonstrated to improve and promote the growth of host plants as well as to reduce disease symptoms caused by plant pathogens through various mechanisms, including the production of secondary metabolites, which are used in direct antagonism against pests and diseases (9, 10, 12), changes in host physiology (42), and the induction of systemic acquired resistance in plants (15). Another significant function found for these actinobacteria was antibiotic activity, suggesting that endophytic actinobacteria can be an interesting source for bioprospecting. New antibiotics from endophytic Streptomyces spp.—alnumycin, munumbicins A to D, and coronamycins—have been reported (7, 11). Recently, two novel antitumor anthraquinones, lupinacidins A and B, were isolated from a new endophytic Micromonospora sp. (43). Moreover, new species of endophytic actinobacteria have been increasingly reported (25, 35). Thus, endophytic actinobacteria are expected to be potential sources of new species and new bioactive agents.Of the myriad ecosystems on earth, those with the greatest general biodiversity seem also to have the greatest number and the greatest diversity of endophytes (73). Tropical and temperate rain forests are the most biologically diverse terrestrial ecosystems on earth and thus the greatest possible resource for the acquisition of novel microorganisms and their products (73). One area of enormous plant biodiversity is Xishuangbanna, located in the People''s Republic of China at the border with Myanmar. This area lies at the ecotone between the Asian tropics and subtropics and is dominated by tropical seasonal rain forests (87). Xishuangbanna contains more than 5,000 species of vascular plants, comprising 16% of China''s total plant diversity, and more than 3,000 are endemic species (53, 60), many of which have ethnobotanical histories. Until the present, little research was carried out to isolate endophytic actinobacteria and their secondary metabolites from Xishuangbanna (36, 86). In our long-term study of endophytic actinobacterial diversity and bioactive metabolites from tropical rain forest medicinal plants in Xishuangbanna, many bioactive endophytic Streptomyces spp. have been isolated (49). However, the work to date is insufficient to provide a general understanding of the diversity, distribution, and ecology of tropical rain forest endophytic actinobacteria and to facilitate further exploitation of the diverse functions of this novel microbial source.In the present study, the diversity of rare endophytic actinobacteria associated with medicinal plants from the tropical rain forest in Xishuangbanna was investigated by combining special culturing techniques. The selected isolates were also identified by 16S rRNA gene analysis. The overall aims of this study were (i) to analyze the actinobacterial community and reveal whether the rain forest investigated in Xishuangbanna represents a valuable source for abundant endophytic actinobacteria and new species, (ii) to evaluate the antimicrobial activities of these actinobacteria and the biosynthetic potential of related secondary metabolites, and (iii) to study the relationships between the taxa of these endophytic actinobacteria and the isolation methods applied.     

家鸽小肠绒毛微血管铸型的扫描电镜观察

用扫描电镜观察了ＡＢＳ丁酮溶液灌注的家鸽小肠绒告发同血管构筑情况。家鸽小肠绒毛血管丛由输入沁动脉、毛细血管网和输出小静脉组成,小肠绒毛血管丰富,并相到吻合成单层密集网;办入小动脉既可从肠腺周围血管丛发出,也可直接由粘膜下去一发出,绒毛下部血管表现为微直血管形态,可能部分具有门静脉性质。     

黄瓜线粒体DNA类质粒pC1的性质和核酸序列研究

津研四号黄瓜线粒体中除主环ＤＮＡ外,还有４种ＤＮＡ类质粒：ｐＣ１、ｐＣ２、ｐＣ３、ｐＣ４。将环形类质粒ｐＣ１ｌｐｋ ｇｃ ｐＵＣ１９的ＥｃｏＲⅠ位点上,克隆至Ｅ．ｃｏｌｉ ＪＭ１０９中。以克隆的ｐＣ１为探针,进行同源性检测,ｐＣ１与津研四号黄瓜的核基因组、叶绿体基因组、线粒体基因组和线粒体中其他类质粒不同源。对ｐＣ１进行序列测定和分析,ｐＣ１长度２ ８８９ｂｐ,含有多个正向和反向重复序列,有３个８