淫羊藿属植物的数量分类学研究

通过观测淫羊藿属植物的30个质量性状和15个数量性状,利用聚类分析和主成分分析的方法研究淫羊藿属属下类群的分类关系。结果表明:(1)聚类分析结果将淫羊藿属中国种类划分为大花类群和小花类群,支持Stearn对Sect.Macroceras、Sect.Polyphyllon和Subg.Rhizophyllum的处理,但认为Sect.Epimedium的分类地位尚需进一步探讨。(2)主成分分析结果显示,性状的累积贡献率不是很高,前3个主成分累积贡献率为51.86%,这可能与本属植物演化过程中性状变异的多样化和复杂化相关,但由主成分分析的结果仍可以看出中国种类被划分为大花类群和小花类群。研究认为,花瓣与内萼片长度比、花瓣是否具距、萼片轮数等作为主成分反映的性状对淫羊藿属分类具有重要价值。     

西藏高寒草原苔藓植物群丛特征及其与生态因子的关系

苔藓植物独特的形态结构和生理特征，使其能够生长在极端干旱和寒冷的严酷环境中，苔藓植物是青藏高原高寒草地的植被组成成分之一，发挥着重要的生态作用。为探讨高寒草原苔藓植物的群丛特征，明确影响苔藓植物群丛分布的主要生态因子，本研究在西藏高寒草原区域共设置40个样地，通过样方调查，采集、记录苔藓植物，基于物种和样地生态因子数据，应用双向指示种分析(TWINSPAN)和典范对应分析(CCA)方法对苔藓植物群丛进行数量分类与排序。共记录9科18属36种藓类植物。在科的水平，以丛藓科为主；在属的水平，以对齿藓属(Didymodon)和真藓属(Bryum)为主。优势种分别是短叶对齿藓(Didymodon tectorus)、细叶石灰藓(Barbula gracilenta)、北地对齿藓(Didymodon fallax)和密歇根对齿藓(Didymodon michiganensis)。生活型以丛集型为主，占83.33%。双向指示种分析将苔藓植物群丛分成8个类型，分别是垂蒴真藓(Bryum uliginosum)群丛、北地对齿藓+金黄银藓(Anomobryum auratum)群丛、短叶对齿藓+北地对齿...     

中国薹草属黑穗薹草组的数量分类研究

对中国薹草属黑穗薹草组Sect.Racemosae及外类群冻原薹草组Sect.Frigidae共20种3变种采用38个形态特征进行了数量分类研究。聚类分析结果表明,黑穗薹草组应是一个自然分类群;支持了《中国植物志》将两对相似种：乌拉草（Carer meyeriana）与红原薹草（Carer hongyuanensis）、膨囊薹草（Carex lehmanii）与五台山薹草（Carex montis-wutaii）分别处理为种的观点。主成分分析结果表明,本组的分类特征较为稳定与集中,本组主成分分析散点图与聚类分析的分类结果大致吻合,并表明有关小坚果、果囊、鳞片、主茎叶和苞片的性状在本组的分类中起到了主要作用。     

北京东灵山苔藓植物区系研究

在野外调查和采集标本的基础上,对北京东灵山苔藓植物区系成分及特点进行研究。结果显示,该地区共有苔藓植物27科64属150种(含2变种),其中32种为北京苔藓植物新记录。优势科为青藓科(Brachytheciaceae)、丛藓科(Pottiaceae)、灰藓科(Hypnaceae)、真藓科(Bryaceae)和绢藓科(Entodontaceae)5科;优势属为青藓属(Brachythecium)、绢藓属(Entodon)、紫萼藓属(Grimmia)、真藓属(Bryum)、金灰藓属(Pylaisia)、凤尾藓属(Fissidens)、美喙藓属(Eurhynchium)和毛口藓属(Trichostomum)8属。区系分析表明,东灵山苔藓植物区系地理成分以北温带成分为主(占36.7%)、东亚成分次之(占18.0%)。通过与我国15个地区苔藓植物区系谱主成分分析发现,东灵山苔藓植物区系成分与雾灵山、小五台山与贺兰山相近,与云蒙山最为接近。     

广西那坡县苔藓植物初步研究

通过对广西那坡县475份苔藓植物标本的整理和鉴定,获得该地区苔藓植物40科81属163种及亚种、变种,其中有34个广西新纪录种.在科属组成分析中,优势科有灰藓科、丛藓科、蔓藓科等,优势属为真藓属、灰藓属、青藓属等.区系成分以东亚成分为主,占总数的30.06％,温带和热带成分所占比例也较高,反映那坡县具南北过渡的特点;中...     

山东昆嵛山苔藓植物多样性及区系特征

通过对采自山东昆嵛山的4000余号苔藓植物标本进行鉴定,发现昆嵛山分布有苔藓植物56科131属318种,包括山东新记录种69种。在昆嵛山的苔藓植物中,苔类植物有22科28属41种,藓类植物有34科103属277种。昆嵛山苔藓植物区系主要由青藓科（45种）、真藓科（33种）、丛藓科（31种）、灰藓科（24种）、绢藓科（18种）、曲尾藓科（15种）等一些大科组成。单种属和寡种属数量较多,但特有现象不明显。土生和石生群落优势明显,水生和树生群落缺乏。区系地理成分复杂,分布类型多样,温带成分优势明显,但也具有一定的热带成分。本研究旨在进一步促进山东苔藓植物研究,并与现代苔藓植物学的发展接轨,也为中国苔藓植物的研究提供新的资料。     

贵州乌江东风水库库区消落带苔藓植物区系分析

采用典型调查与路线调查相结合的方法,对贵州乌江东风水库库区消落带苔藓植物区系的物种组成、生活型、分布区类型和丰富性进行了调查和分析.结果表明:该区域共有苔藓植物18科58属101种,其中,藓类植物有16科56属99种,苔类植物有2科2属2种;优势科为灰藓科(Hypnaceae)、青藓科(Brachytheciaceae)和丛藓科(Pottiaceae),优势属为青藓属(Brahchythecium B.S.G.)、真藓属(Bryum Hedw.)和小曲尾藓属〔Dicranella(Müll.Hal.)Schimp.〕;单属科和单种属所占比例均较高,分别占该区域苔藓植物总科数和总属数的500%和638%.该区域分布的苔藓植物生活型可分为交织型、丛集型、垫状和平铺型4类,以丛集型种数最多(48种),占该区域苔藓植物总种数的475%.该区域分布的苔藓植物可划分为12个分布区类型,其中,温带成分种类最多,所占比例为815%;热带成分所占比例仅为185%;中国特有种所占比例也较高,为207%.该区域苔藓植物的丰富性综合系数(Si)为-03608,低于相邻的六冲河下游流域.综合分析结果表明:该区域的苔藓植物多样性较为丰富,多数种类具有较强的抗逆性,且丛集型苔藓种类最多,与库区消落带的特殊生境相适应;地理成分以东亚成分和北温带成分为主,总体属温带性质,且中国特有种较多,反映出该区域苔藓植物区系的特殊性和复杂性.     

中国柴胡属药用植物的数量分类研究（Ⅰ）

综合研究了柴胡属植物的形态,解剖和花粉方面的性状,对中国柴胡属药用的１４种,２变种,１变型进行了数量分类研究。根据植物分类学的实际意义,首次提出了中国柴髹属植物的分类系统,分为２亚属,２组;探讨了柴胡属药用植物各种间的亲缘关系及种间,种下单位的分类关系;并通过对分类性状的主成分分析和排序,找出了柴胡属植物分类中所应依据的重要性状。     

陕西天华山自然保护区苔藓植物区系研究

根据调查和参考有关文献,对陕西天华山自然保护区苔藓植物区系进行了初步分析,结果表明：种类丰富,本区共有苔藓植物46科95属180种（包括种下类群）。其中苔类植物14科18属37种。藓类植物32科77属143种;优势科为青藓科、提灯藓科、丛藓科、羽藓科、真藓科、绢藓科和灰藓科,优势属为提灯藓属、真藓属、青藓属、光萼苔属、绢藓属、灰藓属、凤尾藓属、丛本藓属、缩叶藓属、羽藓属和仙鹤藓属;地理成分复杂、多样。区系联系广泛;种的地理成分统计分析表明。温带成分占绝对优势。热带成分也占有一定比例,反映了本区苔藓植物区系的热带亲缘性;东亚成分也占有重要地位,有42种,其中中国-日本分布有15种;中国特有种有16种,中国特有分布中以西南区系成分为主。本区苔藓植物区系隶属于华中地区,但兼有多种成分。体现本区苔藓植物区系的南北过渡特征。     

金川泥炭地两个尺度下苔藓植物间及其与维管植物间的种间关系分析

在金川泥炭地设置1m×1m和25 cm ×25 cm两个尺度的大、小样方进行植被调查,并利用方差比率法、x2检验、Pearson相关分析和Spearman秩相关分析方法,定量分析了9种苔藓植物和30种主要维管植物间的种间关系.偏叶泥炭藓、大泥炭藓和尖叶泥炭藓3种优势苔藓植物彼此呈显著负关联,占领着各自不同的生境;该3种优势苔藓植物同伴生苔藓及维管植物主要呈正相关,形成独特的群落,据此划分出4个生态种组.Spearman秩相关分析表明,大样方741个种对中,正联结种对数占49.53％,明显大于小样方666个种对中的41.14％.样方大小对分析植物个体大小差异悬殊的泥炭地植物群落的种间联结的影响巨大.苔藓与维管植物间以及苔藓植物间的种间联结甚至种间关系应在大、小样方中分别探讨更合适.     

Formation of 15alpha-hydroxyestriol and 15alpha-hydroxyestradiol from C19 15alpha-hydroxylated precursors in human pregnancy.

A mixture of 3H-15alpha-hydroxyandrostenedione and 14C-15alpha-hydroxydehydroisoandrosterone was injected intravenously into two subjects in the third trimester of pregnancy and, in a second study, directly into two fetuses in utero during transfusion for erythroblastosis fetalis. The urine was collected for 4-5 days and steroid conjugates in the urine were hydrolyzed into sulfate and glucosiduronate fractions. From the glucosiduronate fraction 15alpha-hydroxyestriol, 15alpha-hydroxyestradiol, 15alpha-hydroxyandrostenedione and 15alpha-hydroxydehydroisoandrosterone were isolated. No metabolites were identified in the sulfate fraction of the urine. A marked difference was observed in the metabolism of 15alpha-hydroxyandrostenedione and 15alpha-hydroxydehydroisoandrosterone which is dependent on the route of administration of the substrates. Both substrates were converted to 15alpha-hydroxyestriol and 15alpha-hydroxyestradiol, and the 3H/14C ratios and percentage conversions suggest that 15alpha-hydroxyandrostenedione seems to be a better precursor of the urinary 15alpha-hydroxylated estrogens than 15alpha-hydroxydehydroisoandrosterone. The 3H/14C ratios also suggest that 15alpha-hydroxydehydroisoandrosterone was converted to 15alpha-hydroxyestriol via 15alpha-hydroxyandrostenedione, and that the formation of 15alpha-hydroxyestradiol from 15alpha-hydroxydehydroisoandrosterone via 15alpha-hydroxyandrostenedione is a pathway of minor importance. Finally, 15alpha-hydroxydehydroisoandrosterone was recovered from the urine only when the precursors were injected into the maternal circulation. Also, an unknown metabolite containing only 14C was detected in the glucosiduronate fraction of the urine of each subject.     

MHC class I and TCR avidity control the CD8 T cell response to IL-15/IL-15Rα complex

IL-15 operates via a unique mechanism termed transpresentation. In this system, IL-15 produced by one cell type is bound to IL-15Rα expressed by the same cell and is presented to apposing cells expressing the IL-15Rβ/γC complex. We have shown that administering soluble IL-15Rα complexed with IL-15 can greatly enhance IL-15 activity. We now show that the naive CD8 T cell response to exogenous IL-15/IL-15Rα complex is MHC class I dependent. In the absence of β2 microglobulin, naive CD8 T cells scarcely proliferated in response to IL-15/IL-15Rα complex, whereas memory cells proliferated, although to a lesser extent, compared with levels in control mice. The loss of β2m or FcRn slightly reduced the extended half-life of IL-15/IL-15Rα complex, whereas FcRn deficiency only partially reduced the naive CD8 T cell proliferative response to IL-15/IL-15Rα complex. In addition, we demonstrated a link between TCR avidity and the ability of a T cell to respond to IL-15/IL-15Rα complex. Thus, T cells expressing low-avidity TCR responded poorly to IL-15/IL-15Rα complex, which correlated with a poor homeostatic proliferative response to lymphopenia. The inclusion of cognate peptide along with complex resulted in enhanced proliferation, even when TCR avidity was low. IL-15/IL-15Rα complex treatment, along with peptide immunization, also enhanced activation and the migratory ability of responding T cells. These data suggest that IL-15/IL-15Rα complex has selective effects on Ag-activated CD8 T cells. Our findings have important implications for directing IL-15/IL-15Rα complex-based therapy to specific Ag targets and illustrate the possible adjuvant uses of IL-15/IL-15Rα complex.     

Crystal Structure of the interleukin-15.interleukin-15 receptor alpha complex: insights into trans and cis presentation

Interleukin (IL)-15 is a pleiotropic cytokine that plays a pivotal role in both innate and adaptive immunity. IL-15 is unique among cytokines due to its participation in a trans signaling mechanism in which IL-15 receptor alpha (IL-15Ralpha) from one subset of cells presents IL-15 to neighboring IL-2Rbeta/gammac-expressing cells. Here we present the crystal structure of IL-15 in complex with the sushi domain of IL-15Ralpha. The structure reveals that the alpha receptor-binding epitope of IL-15 adopts a unique conformation, which, together with amino acid substitutions, permits specific interactions with IL-15Ralpha that account for the exceptionally high affinity of the IL-15.IL-15Ralpha complex. Interestingly, analysis of the topology of IL-15 and IL-15Ralpha at the IL-15.IL-15Ralpha interface suggests that IL-15 should be capable of participating in a cis signaling mechanism similar to that of the related cytokine IL-2. Indeed, we present biochemical data demonstrating that IL-15 is capable of efficiently signaling in cis through IL-15Ralpha and IL-2Rbeta/gammac expressed on the surface of a single cell. Based on our data we propose that cis presentation of IL-15 may be important in certain biological contexts and that flexibility of IL-15Ralpha permits IL-15 and its three receptor components to be assembled identically at the ligand-receptor interface whether IL-15 is presented in cis or trans. Finally, we have gained insights into IL-15.IL-15Ralpha.IL-2Rbeta.gammac quaternary complex assembly through the use of molecular modeling.     

Soluble Interleukin IL-15Ralpha is generated by alternative splicing or proteolytic cleavage and forms functional complexes with IL-15

Interleukin 15 (IL-15) is a pleiotropic cytokine that is hardly detectable in biological fluids. Here, we show that IL-15 forms functional heterocomplexes with soluble high affinity IL-15 receptor alpha (IL-15Ralpha) chain in mouse serum and cell-conditioned medium, which prevents IL-15 detection by ELISA. We also demonstrate that two soluble IL-15Ralpha (sIL-15Ralpha) sushi domain isoforms are generated through a novel alternative splicing mechanism within the IL-15Ralpha gene. These isoforms potentiate IL-15 action by promoting the IL-15-mediated proliferation of the CTLL cell line and interferon gamma production by murine NK cells, which suggests a role in IL-15 transpresentation. Conversely, a full-length sIL-15Ralpha ectodomain released by tumor necrosis factor-alpha-converting enzyme (TACE)-dependent proteolysis inhibits IL-15 activity. Thus, a dual mechanism of sIL-15Ralpha generation exists in mice, giving rise to polypeptides with distinct properties, which regulate IL-15 function.     

Preparation of specific antisera to 15alpha-hydroxyestrogen 15-N-acetylglucosaminides.

3-(1-Carboxypropyl) ether derivatives of 15alpha-hydroxyestradiol 15-N-acetylglucosaminide (15alpha-OHE2 15NAG) and 15alpha-hydroxyestriol (E4) 15NAG were synthesized and conjugated with bovine serum albumin. Antisera elicited in rabbits possessed high affinity and specificity for the 15alpha-hydroxyestrogen (15alpha-OHEs) 15NAG, exhibiting no significant cross-reactivity with 15alpha-OHEs and their positional isomers such as 16NAG and 17NAG. Enzyme immunoassay methods developed by using the purified antisera and horseradish peroxidase-labeled antigens were applied to the measurement of 15alpha-OHEs 15NAG and E4 15NAG in normal pregnancy urine. We demonstrated for the first time that the conjugation of N-acetylglucosamine to E4 occurs at the C-15alpha position.     

Transposition behavior of IS15 and its progenitor IS15-Δ: Are cointegrates exclusive end products?

We report that the major product of IS15-promoted transposition is a cointegrate. When present in the multicopy plasmid pBR322, IS15 and its progenitor IS15-delta mediate the formation of cointegrates at frequencies of 3.5 X 10(-4) and 2.9 X 10(-5), respectively. We have studied the stability of the cointegrates generated by IS15 and IS15-delta. While these structures are resolved in a rec+ host, they were stable in a rec- host. These observations suggest that neither IS15 nor IS15-delta encode a resolvase and that cointegration is an end product of their transposition process. These properties of IS15-delta and IS15 can explain the transitions from IS15-delta to IS15 and from IS15 to IS15-delta observed in vivo.     

Soluble interleukin-15 receptor alpha (IL-15R alpha)-sushi as a selective and potent agonist of IL-15 action through IL-15R beta/gamma. Hyperagonist IL-15 x IL-15R alpha fusion proteins

Interleukin-15 (IL-15) is crucial for the generation of multiple lymphocyte subsets (natural killer (NK), NK-T cells, and memory CD8 T cells), and transpresentation of IL-15 by monocytes and dendritic cells has been suggested to be the dominant activating process of these lymphocytes. We have previously shown that a natural soluble form of IL-15R alpha chain corresponding to the entire extracellular domain of IL-15R alpha behaves as a high affinity IL-15 antagonist. In sharp contrast with this finding, we demonstrate in this report that a recombinant, soluble sushi domain of IL-15R alpha, which bears most of the binding affinity for IL-15, behaves as a potent IL-15 agonist by enhancing its binding and biological effects (proliferation and protection from apoptosis) through the IL-15R beta/gamma heterodimer, whereas it does not affect IL-15 binding and function of the tripartite IL-15R alpha/beta/gamma membrane receptor. Our results suggest that, if naturally produced, such soluble sushi domains might be involved in the IL-15 transpresentation mechanism. Fusion proteins (RLI and ILR), in which IL-15 and IL-15R alpha-sushi are attached by a flexible linker, are even more potent than the combination of IL-15 plus sIL-15R alpha-sushi. After binding to IL-15R beta/gamma, RLI is internalized and induces a biological response very similar to the IL-15 high affinity response. Such hyper-IL-15 fusion proteins appear to constitute potent adjuvants for the expansion of lymphocyte subsets.     

Overexpression of 15-lipoxygenase-1 induces growth arrest through phosphorylation of p53 in human colorectal cancer cells

To investigate the function of 15-lipoxygenase-1 (15-LOX-1) in human colorectal cancer, we overexpressed 15-LOX-1 in HCT-116 human colorectal cancer cells. Clones expressing the highest levels of 15-LOX-1 displayed reduced viability compared with the HCT-116-Vector control cells. Further, by cell cycle gene array analyses, the cyclin-dependent kinase inhibitor p21WAF1/CIP1 and MDM2 genes were up-regulated in 15-LOX-1-overexpressing cells. The induction of p21(WAF1/CIP1) and MDM2 were linked to activation of p53 by 15-LOX-1, as there was a dramatic induction of phosphorylated p53 (Ser15) in 15-LOX-1-overesxpressing cells. However, the 15-LOX-1 metabolites 13(S)-hydroxyoctadecadienoic acid and 15(S)-hydroxyeicosatetraenoic acid failed to induce phosphorylation of p53 at Ser15, and the 15-LOX-1 inhibitor PD146176 did not inhibit the phosphorylation of p53 at Ser15 in 15-LOX-1-overexpressing cells. Nonetheless, the growth-inhibitory effects of 15-LOX-1 were p53 dependent, as 15-LOX-1 overexpression had no effect on cell growth in p53 (-/-) HCT-116 cells. Finally, treatment of HCT-116-15-LOX-1 cells with different kinase inhibitors suggested that the effects of 15-LOX-1 on p53 phosphorylation and activation were due to effects on DNA-dependent protein kinase. Collectively, these findings suggest a new mechanism to explain the biological activity of 15-LOX-1, where 15-LOX plays a stoichiometric role in activating a DNA-dependent protein kinase-dependent pathway that leads to p53-dependent growth arrest.     

The hinge region between two ubiquitin-like domains destabilizes recombinant ISG15 in solution

Interferon-stimulated gene (ISG) 15 mediates antiviral responses and also is upregulated within the endometrium in response to the developing embryo during early pregnancy. Structurally, ISG15 resembles two ubiquitin domains (30% identical) that are separated by a hinge region. Recombinant (r) bovISG15 is not stable in solution. It was hypothesized that the hinge region contributed to the instability of rbovISG15. Within 24 h of dialysis, rbovISG15 formed complexes as detected by reducing and denaturing SDS-PAGE. However, chemical perturbations of cysteine prevented formation of rbovISG15 complexes over time. Furthermore, a site-directed mutant of rbovISG15 (Cys80Ser) was isomeric and more stable than rbovISG15. Neither wild-type nor mutant rbovISG15 was able to interact with the ISG15 E1 initiating enzyme, UBE1L, in an in vitro pull-down assay. Ovine (ov) ISG15 has three additional amino acids within the hinge region that were hypothesized to increase stability and the degree of interaction with UBE1L because of increased separation of the ubiquitin-like domains. Over time in solution, rovISG15 the level of rovISG15 secondary structure was diminished, whereas the Cys80Ser rovISG15 structure did not change. A GST-Cys80Ser rovISG15 fusion protein had increased structural stability and enhanced protein-protein interaction with UBE1L after dialysis for 48 h, when compared to the GST-rovISG15 fusion protein or rbovISG15. Models of bovISG15, Cys80Ser bovISG15, and ovISG15 were constructed, which confirmed that the hinge region between the two ubiquitin domains destabilizes rbovISG15 in solution.     

Combined IL-15/IL-15Ralpha immunotherapy maximizes IL-15 activity in vivo

IL-15 has substantial potential as an immunotherapeutic agent for augmenting immune responses. However, the activity of IL-15 is mediated by a unique mechanism in which the cytokine is transpresented by cell-bound high-affinity IL-15Ralpha to target cells expressing the IL-15Rbeta and the common gamma-chain. Thus, the efficacy of administered IL-15 alone may be limited by the availability of free IL-15Ralpha. We now show that administration of soluble IL-15/IL-15Ralpha complexes greatly enhanced IL-15 half-life and bioavailability in vivo. Treatment of mice with this complex, but not with IL-15 alone, resulted in robust proliferation of memory CD8 T cells, NK cells, and NK T cells. The activity of the complex required IL-15Rbeta, but not IL-15Ralpha, expression by the responding cells and was IL-7-independent. Interestingly, IL-15/IL-15Ralpha immunotherapy also caused naive CD8 T cell activation and development into effector cells and long-term memory T cells. Lastly, complexed IL-15, as compared with IL-15 alone, dramatically reduced tumor burden in a model of B16 melanoma. These findings hold significant importance for the use of IL-15 as a potential adjuvant/therapeutic and inducer of homeostatic proliferation, without the necessity for prior immunodepletion.