S100A9参与介导具核梭杆菌促结肠癌细胞的增殖与迁移的作用

目的:探讨S100A9在具核梭杆菌(Fusobacterium nucleatum,Fn)促结肠癌HCT116和SW480细胞增殖与迁移中的作用。方法:Fn感染HCT116和SW480细胞后,分别采用CCK8实验及Transwell实验检测结肠癌细胞的增殖及迁移能力的变化,采用Real time PCR及Western blot分别检测S100A9基因及蛋白质水平的变化;用S100A9 siRNA转染HCT116和SW480细胞后感染Fn,采用CCK8实验及Transwell实验分别检测两株细胞增殖及迁移能力的变化;利用NF-κB抑制剂BAY 11-7082预处理HCT116和SW480细胞后再感染Fn,采用Real time PCR及Western blot分别检测S100A9基因及蛋白质水平的变化。结果:(1) Fn可促进HCT116和SW480细胞增殖及迁移(P 0.001);(2) Fn感染的HCT116和SW480细胞中S100A9基因及蛋白质水平均较相应对照组明显升高,差异均具有高度显著性(P 0. 01或P 0. 001),即Fn上调S100A9的表达;(3)用S100A9 siRNA下调HCT116和SW480细胞中S100A9表达后,Fn促进这两株细胞增殖及迁移的作用则明显减弱,差异均具有高度显著性(P 0. 001),表明S100A9参与介导Fn的促HCT116和SW480细胞增殖及迁移的作用;(4)在NF-κB通路抑制剂BAY 11-7082预处理的HCT116和SW480细胞组,Fn上调S100A9基因及蛋白质水平的作用明显被逆转,差异均具有高度显著性(P 0. 01或P 0. 001),表明激活NF-κB转录活性是Fn上调S100A9的机制之一。结论:Fn可上调S100A9表达且与NF-κB活化有关,上调S100A9是Fn促结肠癌细胞增殖及迁移的机制之一。     

S100A9参与乙型肝炎病毒X蛋白介导的HepG2细胞增殖与迁移

目的:探讨S100A9在乙型肝炎病毒X(HBx)介导的HepG2细胞增殖及迁移中的作用。方法:用表达HBx蛋白的重组腺病毒AdHBx感染HepG2细胞后,用CCK-8实验检测细胞增殖能力及划痕愈合实验检测细胞迁移能力;在HepG2/AdHBx细胞中转染S100A9-siRNA及其对照siRNA后,检测HepG2细胞增殖及迁移能力;在HepG2/Ad HBx和对照组HepG2/AdGFP细胞中,采用Real-time PCR及Western Blot检测S100A9基因及蛋白的表达情况;在HepG2/AdHBx细胞中,加入不同剂量的NF-κB抑制剂BAY11-7082后,检测各组中S100A9的基因及蛋白表达情况。结果:HBx促进HepG2细胞的增殖与迁移; S100A9-siRNA抑制S100A9的表达后,HBx促进HepG2细胞的增殖与迁移的作用降低,HBx介导的HepG2细胞的增殖与迁移部分依赖于S100A9; S100A9基因及蛋白表达在HepG2/AdHBx中较对照组HepG2/Ad GFP显著升高,HBx可致S100A9表达增加;抑制NF-κB转录活性后,AdHBx+BAY11-7082组S100A9基因及蛋白表达较对照组显著降低,阻断NF-κB转录活性可部分抑制HBx调控的S100A9表达。结论:HBx可调控S100A9的表达且与NF-κB活化有关,S100A9参与HBx介导的HepG2细胞的增殖与迁移。     

长链非编码RNA SNHG3对人乳腺癌细胞MCF-7增殖、迁移与侵袭的影响 *

目的:探讨长链非编码RNA SNHG3对人乳腺癌细胞MCF-7增殖、迁移与侵袭的影响。方法:构建SNHG3过表达质粒,实验分别设置阴性对照组(pcDNA-3.1+)与SNHG3基因过表达组(pcDNA-3.1+/SNHG3)。将MCF-7细胞转染对照组质粒和SNHG3过表达质粒,采用实时定量PCR 方法检测 SNHG3 mRNA 转录水平,Western blot 检测MMP9及EMT相关蛋白质水平;集落形成实验检测MCF-7细胞增殖能力;划痕愈合实验检测MCF-7细胞横向迁移能力; Transwell 小室实验检测MCF-7细胞纵向迁移能力及侵袭能力。结果:过表达SNHG3后,MCF-7细胞中SNHG3的mRNA水平显著增高(P<0.001);MCF-7细胞的体外增殖能力明显增加(P<0.01),迁移(P<0.01)与侵袭能力(P<0.001)也显著增强,实时定量PCR, Western blot 结果显示SNHG3可激活EMT相关通路。结论:过表达SNHG3可能通过激活EMT通路促进乳腺癌MCF-7细胞的增殖,迁移与侵袭。     

CXC趋化因子配体8促进结直肠癌微环境中M2型巨噬细胞趋化及浸润

CXC趋化因子配体8(CXC chemokine ligand 8,CXCL8)在结直肠癌等多种肿瘤中高表达,并促进肿瘤恶性进展。研究发现,结直肠癌微环境中有大量M2型巨噬细胞浸润,但CXCL8是否影响M2型巨噬细胞的浸润及其潜在机制尚未可知。本文旨在探讨CXCL8对结直肠癌中M2型巨噬细胞浸润及趋化作用的影响。本研究首先分析了TCGA数据库结直肠癌样本中CXCL8表达水平及免疫细胞浸润情况,并在临床组织中进行验证。随后Western 印迹及qRT-PCR检测5种结直肠癌细胞株CXCL8的表达情况。佛波酯(PMA)及IL-4诱导THP-1至M2型巨噬细胞后,与HCT116、SW480细胞及过表达CXCL8的HCT116(CXCL8/HCT116)、SW480(CXCL8/SW480)共培养,检测M2型巨噬细胞趋化情况。白细胞介素1β(IL-1β)处理HCT116、SW480细胞,检测CXCL8表达情况,与M2型巨噬细胞共培养,分析趋化结果。结果显示,患者癌组织CXCL8表达高于癌旁组织,CXCL8高表达癌组织中存在更多M2型巨噬细胞浸润;IL-1β作用于HCT116或SW480后,CXCL8的mRNA及蛋白质表达水平升高(P<0.05)。Transwell实验证实,CXCL8趋化M2型巨噬细胞(P<0.05)。综上所述,结直肠癌细胞中CXCL8可由IL-1β诱导产生,CXCL8表达增加能够促进M2型巨噬细胞的趋化,结直肠癌微环境中M2型巨噬细胞大量浸润可能与CXCL8表达升高有关。     

过表达miR-31对结肠炎模型小鼠TLR4/NF-κB信号通路及凋亡蛋白的调控

目的: 探讨miR-31对DSS诱发结肠炎小鼠TLR4/NF-κB信号通路和凋亡相关蛋白的影响。方法: ①小鼠结肠炎实验:用1%葡聚糖硫酸钠(DSS)诱发小鼠溃疡性结肠炎(UC)。14只FVB非转基因小鼠随机分为control组(n=6),DSS组(n=8),16只FVB miR-31转基因小鼠随机分为miR-31过表达组(n=8),miR-31过表达+DSS 组(n=8),DSS溶于水后通过饮水给予小鼠。DSS组和miR-31+DSS组第一周饮用1%DSS水,第二周饮用正常无菌水,第三周饮用1%DSS水,如此5周后造模完成,之后留取小鼠的结肠组织,通过Western blot和IHC检测小鼠结肠组织NF-κB p65、TLR4、Bax、Bcl-2蛋白的表达;TUNEL检测小鼠结肠组织细胞凋亡。②细胞培养实验:在人结肠上皮细胞系HCT 116细胞中通过脂质体转染的方法转染miR-31 mimic和inhibitor,使miR-31过表达或敲低,每组均进行三次重复,48 h后收取细胞,通过Western blot检测NF-κB p65、TLR4蛋白的表达。结果: ①动物实验中,与control组相比,小鼠结肠组织中DSS组和miR-31过表达组NF-κB p65、TLR4蛋白表达水平和凋亡细胞指数均显著升高(P＜0.05或P＜0.01),Bcl-2/Bax比值显著降低(P＜0.05或P＜0.01);且与DSS组相比,miR-31+DSS组NF-κB p65、TLR4蛋白表达水平和凋亡细胞指数也显著升高(P＜0.01),Bcl-2/Bax比值显著降低(P＜0.01)。②细胞实验中,与control组相比, HCT 116细胞过表达miR-31组的NF-κB p65、TLR4蛋白表达水平均显著升高(P＜0.05或P＜0.01),敲低miR-31组的NF-κB p65、TLR4蛋白表达水平下降(P＜0.05)。结论: miR-31通过促进TLR4/NF-κB信号通路和介导肠上皮细胞凋亡促进结肠炎的发展。     

S100A9对宫颈癌Hela细胞的增殖和迁移的影响及其机制探讨

为初步探讨S100A9在宫颈癌中的生物学作用,利用携带有人S100A9基因的腺病毒(Adh S100A9)感染于低表达S100A9的宫颈癌Hela细胞,通过MTT法检测细胞增殖能力,划痕愈合试验和Transwell试验检测细胞迁移能力,通过倒置显微镜观察细胞形态变化,采用RT-PCR和蛋白质印迹法检测上皮细胞标志蛋白E-钙黏着蛋白(E-cadherin,E-cad)、间质细胞标志蛋白波形蛋白(vimentin,Vim)及Wnt/β-联蛋白(β-catenin,β-cat)信号通路相关分子的表达。结果显示,与对照组相比,过表达S100A9的Hela细胞增殖活性增强、迁移率增高,细胞形态由"铺路石"样向"梭形"转变,排列紊乱,伴随E-cad降低(P0.05)和Vim升高(P0.01);同时,过表达S100A9的Hela细胞中Wnt/β-cat信号通路的关键分子β-联蛋白水平增加(P0.01),并且入核增多(P0.01),该通路的下游靶基因c-myc、Snail及Twist表达也明显上调。该研究结果提示,S100A9促进宫颈癌Hela细胞的增殖、迁移和上皮–间质转化(epithelial-mesenchymal transition,EMT),激活Wnt/β-cat信号通路;S100A9促进增殖和迁移作用的机制可能与其促进EMT和激活Wnt/β-cat信号通路相关。     

甘草次酸对大肠癌LoVo细胞增殖和侵袭的影响*

目的: 研究不同浓度的甘草次酸对大肠癌LoVo细胞增殖和侵袭的影响。方法: 将大肠癌LoVo细胞分为对照组,甘草次酸低、中、高剂量组(甘草次酸浓度分别为50, 100, 200 μmol/L)和5氟尿嘧啶组(5氟尿嘧啶浓度为100 μmol/L ),各组细胞经过药物孵育24和48 h后进行检测。通过四氮唑蓝试验检测甘草次酸对各组细胞增殖率的影响;通过Annexin V/PI双标流式细胞术检测各组细胞的凋亡率;Transwell 小室法检测各组细胞的侵袭能力;通过蛋白质印迹法检测检各组细胞的NF-κB蛋白表达。结果: 与对照组相比,甘草次酸中、高剂量组和5氟尿嘧啶组中细胞抑制率均显著降低(P＜0.05);甘草次酸中、高剂量组和5氟尿嘧啶组中细胞凋亡率均显著升高(P＜0.05);甘草次酸中、高剂量组和5氟尿嘧啶组中大肠癌LoVo 细胞侵袭能力显著降低(P＜ 0.05);甘草次酸中、高剂量组和5氟尿嘧啶组中NF-κB相对表达量均显著降低(P＜0.05)。结论: 浓度为100 μmol/L和200 μmol/L甘草次酸可以抑制大肠癌LoVo细胞的增殖,降低侵袭能力,这些作用的机制与抑制NF-κB蛋白的表达有关。     

S100A6通过招募和活化巨噬细胞促进血管形成*

目的:探讨S100A6经由巨噬细胞介导的促血管生成作用及机制。方法:(1)用重组蛋白 GST-hS100A6处理巨噬细胞后:①收集上清制备条件培养基(简称A6-Mφ-CM),并用之重悬人脐静脉内皮细胞(HUVEC),用体外血管形成试验检测各处理因素对血管形成的影响;②分别用实时荧光定量PCR和Western blot检测巨噬细胞的M2型标志物CD163及促血管形成因子CCL2、IL-6、VEGFA的mRNA和蛋白质水平,以及JAK2和STAT3的蛋白质及其磷酸化水平;③用Transwell迁移试验检测巨噬细胞迁移能力的变化。(2)使用JAK2抑制剂(XL019)预处理巨噬细胞后再加GST-hS100A6处理,检测S100A6促巨噬细胞迁移作用的变化。以重组蛋白GST为实验对照。 结果:(1)A6-Mφ-CM组的血管分支数和血管分支长度既明显高于GST-Mφ-CM组(P值均小于0.05),也明显高于GST-hS100A6直接处理的HUVEC组(P<0.001,P<0.01),提示S100A6处理后的巨噬细胞具有促进血管形成的作用;(2)GST-hS100A6处理后的巨噬细胞中,CD163、CCL2、IL-6、VEGFA的mRNA和蛋白质水平明显高于GST组(P值均小于0.05),提示S100A6诱导巨噬细胞向促血管表型(pro-angiogenic phenotype)转化;(3)GST-hS100A6处理后,巨噬细胞的迁移数是GST组的1.4倍(P<0.01),提示S100A6具有招募巨噬细胞的作用;(4)GST-hS100A6处理组巨噬细胞的JAK2和STAT3的蛋白质及其磷酸化水平都明显高于GST组(P值均小于0.05),而JAK2抑制剂XL019可部分抑制S100A6促进巨噬细胞迁移的作用(P<0.01),提示S100A6促进巨噬细胞迁移作用机制涉及JAK2/STAT3信号通路的激活。 结论:微环境中的S100A6可通过招募巨噬细胞并进一步诱导其向促血管表型转化,进而促进新生血管形成;其招募巨噬细胞的机制涉及JAK2/STAT3信号通路的激活。     

低氧促进肺腺癌A549细胞迁移的机制*

目的: 探讨低氧促进肺腺癌A549细胞迁移的机制。方法: 培养肺腺癌A549细胞,转染慢病毒获得稳定敲低ACC1的A549细胞株,转染si-RNA获得敲低SREBP-1的A549细胞。分别以低氧(5％ O₂)联合低氧诱导因子1α(HIF-1α)抑制剂PX-478(25 μmol)处理A549细胞,低氧联合亚油酸(LA)(20 μmol)处理敲低ACC1的A549细胞,低氧处理敲低胆固醇调节原件结合蛋白1(SREBP-1)的A549细胞。Transwell实验检测细胞迁移,蛋白质印迹法检测HIF-1α、ACC1及上皮-间质转化(EMT)相关波形蛋白(Vimentin)及E-钙黏蛋白(E-Cadherin)的表达与SREBP-1的表达,实时荧光定量聚合酶链反应(RT-qPCR)检测低氧联合HIF-1α抑制剂PX-478(25 μmol)处理A549细胞后ACC1及SREBP-1 mRNA水平变化。每项实验重复三次。结果: 与常氧组相比,低氧组A549细胞迁移数增加,ACC1与HIF-1α表达上调(P均＜0.01),SREBP-1表达上调(P＜0.05);与低氧对照组相比,PX-478(25 μmol)抑制A549细胞迁移,SREBP-1表达下调(P＜0.05);低氧处理A549细胞后ACC1 mRNA上升(P＜0.05),SREBP-1 mRNA水平上升(P＜0.01);低氧并使用PX-478(25 μmol)处理A549细胞24 h,ACC1 mRNA水平下降(P＜0.05),SREBP-1 mRNA 水平下降(P＜0.01);转染si-RNA获得敲低SREBP-1的A549细胞,Transwell 实验显示si-SREBP-1组细胞迁移数较常氧对照组减少(P＜0.01);低氧处理si-SREBP-1组与si-NC组,与对照组相比si-SREBP-1组细胞迁移数减少(P＜0.01)但与常氧组相比差异无统计学意义(P＞0.05);Western blot检测到si-SREBP-1组ACC1表达较对照组下降(P＜0.01);低氧处理si-SREBP-1组,ACC1表达较对照组下降(P＜0.01);敲低ACC1抑制A549细胞迁移(P＜0.05),敲低ACC1后A549细胞在常氧和5％ O₂条件下细胞迁移数目差异无统计学意义(P＞ 0.05);低氧处理敲低ACC1的A549细胞并给予LA(25 μmol)促进A549细胞迁移(P＜0.05)。结论: 低氧通过HIF-1α/SREBP-1/ACC1途径调节脂肪酸代谢进而促进肺腺癌A549细胞迁移。     

Agroinfiltration and PVX Agroinfection in Potato and Nicotiana benthamiana

Agroinfiltration and PVX agroinfection are two efficient transient expression assays for functional analysis of candidate genes in plants. The most commonly used agent for agroinfiltration is Agrobacterium tumefaciens, a pathogen of many dicot plant species. This implies that agroinfiltration can be applied to many plant species. Here, we present our protocols and expected results when applying these methods to the potato (Solanum tuberosum), its related wild tuber-bearing Solanum species (Solanum section Petota) and the model plant Nicotiana benthamiana. In addition to functional analysis of single genes, such as resistance (R) or avirulence (Avr) genes, the agroinfiltration assay is very suitable for recapitulating the R-AVR interactions associated with specific host pathogen interactions by simply delivering R and Avr transgenes into the same cell. However, some plant genotypes can raise nonspecific defense responses to Agrobacterium, as we observed for example for several potato genotypes. Compared to agroinfiltration, detection of AVR activity with PVX agroinfection is more sensitive, more high-throughput in functional screens and less sensitive to nonspecific defense responses to Agrobacterium. However, nonspecific defense to PVX can occur and there is a risk to miss responses due to virus-induced extreme resistance. Despite such limitations, in our experience, agroinfiltration and PVX agroinfection are both suitable and complementary assays that can be used simultaneously to confirm each other''s results.     

Mechanisms and ecological consequences of plant defence induction and suppression in herbivore communities

Background Plants are hotbeds for parasites such as arthropod herbivores, which acquire nutrients and energy from their hosts in order to grow and reproduce. Hence plants are selected to evolve resistance, which in turn selects for herbivores that can cope with this resistance. To preserve their fitness when attacked by herbivores, plants can employ complex strategies that include reallocation of resources and the production of defensive metabolites and structures. Plant defences can be either prefabricated or be produced only upon attack. Those that are ready-made are referred to as constitutive defences. Some constitutive defences are operational at any time while others require activation. Defences produced only when herbivores are present are referred to as induced defences. These can be established via de novo biosynthesis of defensive substances or via modifications of prefabricated substances and consequently these are active only when needed. Inducibility of defence may serve to save energy and to prevent self-intoxication but also implies that there is a delay in these defences becoming operational. Induced defences can be characterized by alterations in plant morphology and molecular chemistry and are associated with a decrease in herbivore performance. These alterations are set in motion by signals generated by herbivores. Finally, a subset of induced metabolites are released into the air as volatiles and function as a beacon for foraging natural enemies searching for prey, and this is referred to as induced indirect defence.Scope The objective of this review is to evaluate (1) which strategies plants have evolved to cope with herbivores and (2) which traits herbivores have evolved that enable them to counter these defences. The primary focus is on the induction and suppression of plant defences and the review outlines how the palette of traits that determine induction/suppression of, and resistance/susceptibility of herbivores to, plant defences can give rise to exploitative competition and facilitation within ecological communities “inhabiting” a plant.Conclusions Herbivores have evolved diverse strategies, which are not mutually exclusive, to decrease the negative effects of plant defences in order to maximize the conversion of plant material into offspring. Numerous adaptations have been found in herbivores, enabling them to dismantle or bypass defensive barriers, to avoid tissues with relatively high levels of defensive chemicals or to metabolize these chemicals once ingested. In addition, some herbivores interfere with the onset or completion of induced plant defences, resulting in the plant’s resistance being partly or fully suppressed. The ability to suppress induced plant defences appears to occur across plant parasites from different kingdoms, including herbivorous arthropods, and there is remarkable diversity in suppression mechanisms. Suppression may strongly affect the structure of the food web, because the ability to suppress the activation of defences of a communal host may facilitate competitors, whereas the ability of a herbivore to cope with activated plant defences will not. Further characterization of the mechanisms and traits that give rise to suppression of plant defences will enable us to determine their role in shaping direct and indirect interactions in food webs and the extent to which these determine the coexistence and persistence of species.     

The diversity,ecology and evolution of extrafloral nectaries: current perspectives and future challenges

Background

Plants in over one hundred families in habitats worldwide bear extrafloral nectaries (EFNs). EFNs display a remarkable diversity of evolutionary origins, as well as diverse morphology and location on the plant. They secrete extrafloral nectar, a carbohydrate-rich food that attracts ants and other arthropods, many of which protect the plant in return. By fostering ecologically important protective mutualisms, EFNs play a significant role in structuring both plant and animal communities. And yet researchers are only now beginning to appreciate their importance and the range of ecological, evolutionary and morphological diversity that EFNs exhibit.

Scope

This Highlight features a series of papers that illustrate some of the newest directions in the study of EFNs. Here, we introduce this set of papers by providing an overview of current understanding and new insights on EFN diversity, ecology and evolution. We highlight major gaps in our current knowledge, and outline future research directions.

Conclusions

Our understanding of the roles EFNs play in plant biology is being revolutionized with the use of new tools from developmental biology and genomics, new modes of analysis allowing hypothesis-testing in large-scale phylogenetic frameworks, and new levels of inquiry extending to community-scale interaction networks. But many central questions remain unanswered; indeed, many have not yet been asked. Thus, the EFN puzzle remains an intriguing challenge for the future.     

Unraveling regulation and new components of human P-bodies through a protein interaction framework and experimental validation

The cellular factors involved in mRNA degradation and translation repression can aggregate into cytoplasmic domains known as GW bodies or mRNA processing bodies (P-bodies). However, current understanding of P-bodies, especially the regulatory aspect, remains relatively fragmentary. To provide a framework for studying the mechanisms and regulation of P-body formation, maintenance, and disassembly, we compiled a list of P-body proteins found in various species and further grouped both reported and predicted human P-body proteins according to their functions. By analyzing protein-protein interactions of human P-body components, we found that many P-body proteins form complex interaction networks with each other and with other cellular proteins that are not recognized as P-body components. The observation suggests that these other cellular proteins may play important roles in regulating P-body dynamics and functions. We further used siRNA-mediated gene knockdown and immunofluorescence microscopy to demonstrate the validity of our in silico analyses. Our combined approach identifies new P-body components and suggests that protein ubiquitination and protein phosphorylation involving 14-3-3 proteins may play critical roles for post-translational modifications of P-body components in regulating P-body dynamics. Our analyses provide not only a global view of human P-body components and their physical interactions but also a wealth of hypotheses to help guide future research on the regulation and function of human P-bodies.     

Plant growth and architectural modelling and its applications. Preface

Over the last decade, a growing number of scientists around the world have invested in research on plant growth and architectural modelling and applications (often abbreviated to plant modelling and applications, PMA). By combining physical and biological processes, spatially explicit models have shown their ability to help in understanding plant–environment interactions. This Special Issue on plant growth modelling presents new information within this topic, which are summarized in this preface. Research results for a variety of plant species growing in the field, in greenhouses and in natural environments are presented. Various models and simulation platforms are developed in this field of research, opening new features to a wider community of researchers and end users. New modelling technologies relating to the structure and function of plant shoots and root systems are explored from the cellular to the whole-plant and plant-community levels.     

Design and Use of Multiplexed Chemostat Arrays

Chemostats are continuous culture systems in which cells are grown in a tightly controlled, chemically constant environment where culture density is constrained by limiting specific nutrients.^1,2 Data from chemostats are highly reproducible for the measurement of quantitative phenotypes as they provide a constant growth rate and environment at steady state. For these reasons, chemostats have become useful tools for fine-scale characterization of physiology through analysis of gene expression^3-6 and other characteristics of cultures at steady-state equilibrium.⁷ Long-term experiments in chemostats can highlight specific trajectories that microbial populations adopt during adaptive evolution in a controlled environment. In fact, chemostats have been used for experimental evolution since their invention.⁸ A common result in evolution experiments is for each biological replicate to acquire a unique repertoire of mutations.^9-13 This diversity suggests that there is much left to be discovered by performing evolution experiments with far greater throughput. We present here the design and operation of a relatively simple, low cost array of miniature chemostats—or ministats—and validate their use in determination of physiology and in evolution experiments with yeast. This approach entails growth of tens of chemostats run off a single multiplexed peristaltic pump. The cultures are maintained at a 20 ml working volume, which is practical for a variety of applications. It is our hope that increasing throughput, decreasing expense, and providing detailed building and operation instructions may also motivate research and industrial application of this design as a general platform for functionally characterizing large numbers of strains, species, and growth parameters, as well as genetic or drug libraries.     

红花水提取液对小鼠系统性硬皮病的防治作用及其机制*

目的:研究红花水提取液对系统性硬皮病(SSc)模型小鼠的防治作用及相关机制研究。方法:60只 BALB /C小鼠随机分为对照组、模型组、强的松组、红花低、中、高剂量组,每组10只。对照组背部注射生理盐水,其余5组均背部皮下注射100 μl浓度为 200 μg /ml的注射用盐酸博来霉素,每天1次,连续注射28 d,制备SSc模型;造模同时对照组和模型组给予生理盐水10 ml/kg灌胃,强的松组给予强的松溶液4.5 mg/kg (10 ml/kg)灌胃,红花低、中、高剂量组分别给予红花1.5、3、6 g/kg (10 ml/kg)灌胃,各组均连续灌胃28 d。给药28 d后,取各组小鼠背部注射博来霉素区皮肤组织切片测量真皮厚度,采用水解法检测皮肤组织羟脯氨酸(HYP)含量;采用ELISA法检测皮肤组织结缔组织生长因子(CTGF)、转化生长因子-β(TGF-β)含量及血清白细胞介素-6(IL-6)、白细胞介素-17(IL-17)水平。结果:与对照组比较,模型组皮肤真皮厚度,皮肤组织CTGF、TGF-β、HYP含量及血清 IL-6、IL-17 水平明显升高(P＜0.05);与模型组比较,强的松组、红花低、中、高剂量组皮肤真皮厚度,皮肤组织 CTGF、TGF-β、HYP含量及血清 IL-6、IL-17水平明显降低(P＜0.05)。结论:红花水提取液可改善SSc小鼠皮肤状况(或真皮厚度),其作用机制可能与减轻免疫炎症反应有关。     

Floral structure and development and systematic aspects of some 'lower' Asparagales

 Floral structure and development of representatives of Asteliaceae, Blandfordiaceae, Boryaceae, Doryanthaceae, and Hypoxidaceae, all members of the `lower' Asparagales, were studied comparatively. The results are discussed in the light of new molecular systematic studies, but also with regard to established morphological characters in related groups. Stamen shape varies considerably within and between taxa: the shape of anthers is from X-shaped, sagittate to non-sagittate, they are either latrorse or introrse, basifixed, centrifixed or dorsifixed. Gynoecia are syncarpous up to the stigmatic region in all taxa. Ovaries of Doryanthaceae and Hypoxidaceae are inferior, but they are superior in Asteliaceae, Blandfordiaceae and Boryaceae. All ovaries have at least a short synascidiate zone. With the exception of Astelia alpina (Asteliaceae), the ovaries are trilocular. Ovaries of Asteliaceae contain mucilage, which is secreted from trichomes on the funicle and on the placenta. Although flowers are polysymmetric at anthesis, they are monosymmetric in earliest stages with a developmental gradient from adaxial to abaxial. Perianth organs arise individually from either a concave (taxa with inferior ovary) or convex (taxa with superior ovary) apex. Hypoxidaceae have pollen flowers with free stamens. One species, Curculigo capitulata, has Solanum-type flowers with postgenitally united stamens. It is most probably pollinated by buzzing bees. All other taxa have nectariferous flowers with internal or external septal nectaries. Received February 5, 2001 Accepted June 20, 2001     

Efficacy and acceptability of pharmacological,psychosocial, and brain stimulation interventions in children and adolescents with mental disorders: an umbrella review

Top‐tier evidence on the safety/tolerability of 80 medications in children/adolescents with mental disorders has recently been reviewed in this jour­nal. To guide clinical practice, such data must be combined with evidence on efficacy and acceptability. Besides medications, psychosocial inter­ventions and brain stimulation techniques are treatment options for children/adolescents with mental disorders. For this umbrella review, we systematically searched network meta‐analyses (NMAs) and meta‐analyses (MAs) of randomized controlled trials (RCTs) evaluating 48 medications, 20 psychosocial interventions, and four brain stimulation techniques in children/adolescents with 52 different mental disorders or groups of mental disorders, reporting on 20 different efficacy/acceptability outcomes. Co‐primary outcomes were disease‐specific symptom reduction and all‐cause discontinuation (“acceptability”). We included 14 NMAs and 90 MAs, reporting on 15 mental disorders or groups of mental disorders. Overall, 21 medications outperformed placebo regarding the co‐primary outcomes, and three psychosocial interventions did so (while seven outperformed waiting list/no treatment). Based on the meta‐analytic evidence, the most convincing efficacy profile emerged for amphetamines, methylphenidate and, to a smaller extent, behavioral therapy in attention‐deficit/hyperactivity disorder; aripiprazole, risperidone and several psychosocial interventions in autism; risperidone and behavioral interventions in disruptive behavior disorders; several antipsychotics in schizophrenia spectrum disorders; fluoxetine, the combination of fluoxetine and cognitive behavioral therapy (CBT), and interpersonal therapy in depression; aripiprazole in mania; fluoxetine and group CBT in anxiety disorders; fluoxetine/selective serotonin reuptake inhibitors, CBT, and behavioral therapy with exposure and response prevention in obsessive‐compulsive disorder; CBT in post‐traumatic stress disorder; imipramine and alarm behavioral intervention in enuresis; behavioral therapy in encopresis; and family therapy in anorexia nervosa. Results from this umbrella review of interventions for mental disorders in children/adolescents provide evidence‐based information for clinical decision making.