Ovarian hormone level alterations during rat post-reproductive life-span influence CD8?+?T-cell homeostasis

The study examined the putative role of ovarian hormones in shaping of rat peripheral T-cell compartment during post-reproductive period. In 20-month-old rats ovariectomized (Ox) at the very end of reproductive period, thymic output, cellularity and composition of major TCRαβ + peripheral blood lymphocyte and splenocyte subsets were analyzed. Ovariectomy led to the enlargement of CD8 + peripheral blood lymphocyte and splenocyte subpopulations. This reflected: (i) a more efficient thymic generation of CD8 + cells as indicated by increased number of CD4+CD8 + double positive and the most mature CD4-CD8+TCRαβ^high thymocytes and CD8 + recent thymic emigrants (RTEs) in peripheral blood, but not in the spleen of Ox rats, and (ii) the expansion of CD8 + memory/activated peripheral blood lymphocytes and splenocytes. The latter was consistent with a greater frequency of proliferating cells among freshly isolated memory/activated CD8 + peripheral blood lymphocytes and splenocytes and increased proliferative response of CD8 + splenocytes to stimulation with plate-bound anti-CD3 antibody. The former could be related to the rise in splenic IL-7 and IL-15 mRNA expression. Although ovariectomy affected the overall number of CD4 + T cells in none of the examined compartments, it increased CD4+FoxP3 + peripheral blood lymphocyte and splenocyte counts by enhancing their generation in periphery. Collectively, the results suggest that ovariectomy-induced long-lasting disturbances in ovarian hormone levels (mirrored in diminished progesterone serum level in 20-month-old rats) affects both thymic CD8 + cell generation and peripheral homeostasis and leads to the expansion of CD4+FoxP3 + cells in the periphery, thereby enhancing autoreactive cell control on account of immune system efficacy to combat infections and tumors.     

MICROBIOLOGY Vo1 .2 8 2 0 0 1 CONTENTS

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Mutators in SACCHAROMYCES CEREVISIAE: MUT1–1, MUT1–2 and MUT2–1

The purpose of this study was to characterize two mutator stocks of yeast which were induced and selected on the basis of high spontaneous reversion rates of the suppressible "ochre" nonsense allele lys1-1. In the mutator stock VA-3, a single mutation, designated mut1-1, is responsible for the increase in the reversion rate of the ochre alleles lys1-1 and arg4-17. In stock VA-105, there are two separate mutator mutations. Tetrad analysis data showed these two loci are loosely linked. Based on complementation data, one of these mutations is at the same locus as mut1-1 and designated mut1-2. The second mutator of stock VA-105 was designated mut2-1. All three mutators are recessive. Both mut1-1 and mut1-2 give a high mutation rate for ochre nonsense suppressor (SUP) loci, but not for the ochre nonsense alleles. On the contrary, the mutation rates of the ochre alleles are greatly reduced. With the mutant mut2-1 there were mutations at both the lys1-1 site and its suppressors; mut2-1 is as effective as mut1-2 but not as effective as mut1-1 in inducing reversions of a missense mutant, his1-7. Neither mut1-1, mut1-2 nor mut2-1 were effective in inducing reversions of a putative frameshift mutation, hom3-10, or in inducing forward mutations to canavanine resistance.     

潘庆民1,于振文2,王月福2

采用盆栽和水泥池栽研究了追氮时期对小麦光合作用、14C同化物运转分配和硝酸还原酶(NR)活性的影响.结果表明,拔节(雌雄蕊原基形成)期较起身(二棱)期追施氮肥,显著提高了小麦开花后的旗叶叶绿素含量和单叶光合速率;灌浆期旗叶14C同化物向籽粒转移比例显著提高,而在营养器官的滞留比例显著降低;旗叶和根系中硝酸还原酶(NR)活性亦显著提高.小麦穗粒数、粒重和产量增加,蛋白质含量提高.     

Dynamic relocation of the TORC1–Gtr1/2–Ego1/2/3 complex is regulated by Gtr1 and Gtr2

TORC1 regulates cellular growth, metabolism, and autophagy by integrating various signals, including nutrient availability, through the small GTPases RagA/B/C/D in mammals and Gtr1/2 in budding yeast. Rag/Gtr is anchored to the lysosomal/vacuolar membrane by the scaffold protein complex Ragulator/Ego. Here we show that Ego consists of Ego1 and Ego3, and novel subunit Ego2. The ∆ego2 mutant exhibited only partial defects both in Gtr1-dependent TORC1 activation and Gtr1 localization on the vacuole. Ego1/2/3, Gtr1/2, and Tor1/Tco89 were colocalized on the vacuole and associated puncta. When Gtr1 was in its GTP-bound form and TORC1 was active, these proteins were preferentially localized on the vacuolar membrane, whereas when Gtr1 was in its GDP-bound form, they were mostly localized on the puncta. The localization of TORC1 to puncta was further facilitated by direct binding to Gtr2, which is involved in suppression of TORC1 activity. Thus regulation of TORC1 activity through Gtr1/Gtr2 is tightly coupled to the dynamic relocation of these proteins.     

Urotensin-2 promotes collagen synthesis via ERK1/2-dependent and ERK1/2-independent TGF-β1 in neonatal cardiac fibroblasts

U2 (urotensin-2) is the most potent vasoconstrictor in mammals which is involved in cardiac remodelling, including cardiac hypertrophy and cardiac fibrosis. Although the cellular mechanisms of the U2-induced vasoconstriction have been extensively studied, the signalling pathways involved in U2-induced TGF-β1 (transforming growth factor-β1) expression and collagen synthesis remain unclear. In this study, we show that U2 promoted collagen synthesis and ERK1/2 (extracellular signal-regulated kinase 1/2) activation in neonatal cardiac fibroblasts. The U2-induced collagen synthesis and TGF-β1 production were significantly but not completely inhibited by blocking ERK1/2. Both ERK1/2 inhibitor and TGF-β1 antibody could separately inhibit U2-induced collagen synthesis, and the synergistic inhibition effect was observed by blocking ERK1/2 and TGF-β1 simultaneously. These data suggest that U2 promotes collagen synthesis via ERK1/2-dependent and independent TGF-β1 pathway in neonatal cardiac fibroblasts.     

Expressivity of apomixis in 2n?+?n hybrids from an apomictic and a sexual parent: insights into variation detected in Pilosella (Asteraceae: Lactuceae)

Reproductive variation was studied in the tetraploid Pilosella aurantiaca, hexaploid P. rubra (both species with facultative autonomous apospory) and in their 2n + n hybrids, which were obtained by crossing with a sexual pollen parent (tetraploid P. officinarum). The different DNA content in P. aurantiaca and P. officinarum demonstrated the actual 2n + n origin, both spontaneous from the field and through experimental crosses, of their hexaploid hybrids. The octoploid 2n + n progeny were recovered from an experimental cross of P. rubra and P. officinarum. The reproductive pathways operating in two maternal facultatively apomictic species and in the hybrids were quantified using a flow cytometric analysis of seeds obtained from either open-pollinated or emasculated plants. Whereas both maternal species displayed a high penetrance of apomixis, the level of apomixis among the majority of 2n + n hybrids was much lower and variable. Some of the hexaploid hybrids had a reduced seed set. Compared to the respective maternal parents, the decrease in apomixis due to haploid parthenogenesis and/or n + n mating was evident in almost all unreduced hybrids, irrespective of their field/experimental origin and ploidy. Hence, the reproductive behaviour in the apomictic maternal parent was profoundly different from that of the 2n + n hybrids with a sexual parent in spite of the preservation of the complete maternal genome in the hybrids. The regulatory interactions in hybrid genomes, such as effects of modifiers, heterochrony, and epigenetic control, may be consistent with the different expressivity of apomixis observed under different genetic backgrounds.     

猪链球菌1、2、14、1/2型荚膜多糖的单糖组成比较

【目的】猪链球菌1、2、14和1/2型间存在单向或双向的交叉抗原性,这种交叉抗原性的产生原因至今未被揭示。【方法】采用Sephacryl S-300凝胶层析柱对猪链球菌14和1/2型荚膜多糖进行分离纯化,经苯酚-硫酸检测和dot-ELISA辅助鉴定,确定荚膜多糖成分。采用高效凝胶渗透色谱法测定14和1/2型猪链球菌荚膜多糖分子量分别为487.38 kDa和512.72 kDa。【结果】经柱前衍生高效液相色谱法、荧光标记液相色谱法和核磁共振测定14和1/2型猪链球菌荚膜多糖单糖组成分别为:Glc/Gal/GlcNAc/Rha/Neu5Ac(1∶2.94∶1.35∶0.24∶0.37)和Glc/Gal/GlcNAc/GalNAc/Rha/Neu5Ac(1∶1.67∶1.05∶0.93∶0.72∶0.7)。并与已知的猪链球菌1、2型荚膜多糖的单糖组成进行比较分析,发现4种血清型荚膜多糖都具有Glc、GlcNAc、Gal和Neu5Ac,但单糖组成和比列并无明显相似性,这种交叉抗原性可能是由于荚膜多糖的空间结构相似性和(或)细胞表面的其他成分引起的。     

ERK1/2和p-ERK1/2在肺癌组织中的表达及意义

目的研究细胞外信号调节激酶1/2（extracellular regulated kinase 1/2,ERK1/2）及其磷酸化状态（p-ERK1/2）在不同分化程度肺癌中的表达情况,探讨二者与肺癌侵袭、转移的关系。方法采用免疫组化（Envision）法,检测79例肺癌组织及l2例癌旁正常肺组织中ERK1/2和p-ERK1/2的表达。结果ERK1/2在高、中、低分化组表达率分别为13.6%,39.4%,66.7%,p-ERK1/2在高、中、低分化组表达率分别14.3%,27.3%,79.2%（P〈0.05）;无淋巴结转移者阳性率为20%,有淋巴结转移者阳性率为50.1%（P〈0.05）。ERK1/2和p-ERK1/2的表达在不同年龄、性别、肿瘤大小、肿瘤病理类型无显著性差异,而与分化程度有关,其中p-ERK1/2的表达还与有无淋巴结转移有关。结论ERK1/2和p-ERK1/2在肺癌组织中高表达且与分化程度有关。     

赵印泉1，2， 周斯建1， 彭培好1， 潘会堂2， 张启翔2*

以三轮玉蝶梅为材料,采用顶空-固相微萃取与气相色谱-质谱联用技术从5个开花阶段、24 h不同时段和花器官不同部位共鉴定了33种挥发性成分,其中苯基/苯丙烷类化合物占优,还有少量脂肪酸衍生物类和萜烯类化合物。三轮玉蝶梅花朵在不同开花阶段和24 h内不同时段释放的挥发性成分的种类、含量和出现频率有较大的差异。在不同开花阶段,苯基/苯丙烷类化合物数量从低 高 低的趋势,脂肪酸衍生物从低 高的趋势,萜烯类化合物从高 低的趋势;在24 h内不同时段,三类化合物释放的节律不同,苯基/苯丙烷类化合物、脂肪酸衍生物和萜烯类化合物分别在2：00、14：00和6：00达到最高,这表明梅花挥发物的释放具有多种调节模式。花器官不同部位挥发性成分的释放具有器官的特异性。     

赵印泉1，2， 周斯建1， 彭培好1， 潘会堂2， 张启翔2*

以三轮玉蝶梅为材料,采用顶空-固相微萃取与气相色谱-质谱联用技术从5个开花阶段、24 h不同时段和花器官不同部位共鉴定了33种挥发性成分,其中苯基/苯丙烷类化合物占优,还有少量脂肪酸衍生物类和萜烯类化合物。三轮玉蝶梅花朵在不同开花阶段和24 h内不同时段释放的挥发性成分的种类、含量和出现频率有较大的差异。在不同开花阶段,苯基/苯丙烷类化合物数量从低-高-低的趋势,脂肪酸衍生物从低-高的趋势,萜烯类化合物从高-低的趋势;在24 h内不同时段,三类化合物释放的节律不同,苯基/苯丙烷类化合物、脂肪酸衍生物和萜烯类化合物分别在2:00、14:00和6:00达到最高,这表明梅花挥发物的释放具有多种调节模式。花器官不同部位挥发性成分的释放具有器官的特异性。     

Regulation of α(2B)-adrenergic receptor-mediated extracellular signal-regulated kinase 1/2 (ERK1/2) activation by ADP-ribosylation factor 1

A number of signaling molecules are involved in the activation of the mitogen-activated protein kinase (MAPK) pathway by G protein-coupled receptors. In this study, we have demonstrated that α(2B)-adrenergic receptor (α(2B)-AR) interacts with ADP-ribosylation factor 1 (ARF1), a small GTPase involved in vesicle-mediated trafficking, in an agonist activation-dependent manner and that the interaction is mediated through a unique double Trp motif in the third intracellular loop of the receptor. Interestingly, mutation of the double Trp motif and siRNA-mediated depletion of ARF1 attenuate α(2B)-AR-mediated activation of extracellular signal-regulated kinases 1/2 (ERK1/2) without altering receptor intracellular trafficking, whereas expression of the constitutively active mutant ARF1Q71L and ARNO, a GDP-GTP exchange factor of ARF1, markedly enhances the activation of Raf1, MEK1, and ERK1/2. These data strongly demonstrate that the small GTPase ARF1 modulates ERK1/2 activation by α(2B)-AR and provide the first evidence indicating a novel function for ARF1 in regulating the MAPK signaling pathway.     

Pkh1/2-Pil1信号调节酵母细胞内吞

酵母细胞中Pkh1/2信号影响细胞内吞功能.Pil1因与Pkh1/2形成复合物并可被 其磷酸化引起关注.新近发现一个大分子复合物eisosome 是内吞的标志性位点， 而Pil1是其主要成分.前期研究发现，Pil1的磷酸化状态对eisosome结构完整至关 重要.本研究通过检测Pkh1/2突变菌的生长和萤光黄（LY） 染料在空泡聚集情况 ，发现由于Pkh1/2突变导致的生长抑制和液相内吞功能丧失,可部分由过表达的 Pil1补偿，得出结论是Pkh1/2-Pil1信号在一定程度上调节细胞内吞.     

Nod2 downregulates TLR2/1 mediated IL1β gene expression in mouse peritoneal macrophages

Nod2 is a cytosolic pattern recognition receptor. It has been implicated in many inflammatory conditions. Its signaling has been suggested to modulate TLR responses in a variety of ways, yet little is known about the mechanistic details of the process. We show in this study that Nod2 knockdown mouse peritoneal macrophages secrete more IL1β than normal macrophages when stimulated with peptidoglycan (PGN). Muramyl dipeptide (MDP, a Nod2 ligand) + PGN co-stimulated macrophages have lower expression of IL1β than PGN (TLR2/1 ligand) stimulated macrophages. MDP co-stimulation have similar effects on Pam3CSK4 (synthetic TLR2/1 ligand) mediated IL1β expression suggesting that MDP mediated down regulating effects are receptor dependent and ligand independent. MDP mediated down regulation was specific for TLR2/1 signaling as MDP does not affect LPS (TLR4 ligand) or zymosan A (TLR2/6 ligand) mediated IL1β expression. Mechanistically, MDP exerts its down regulating effects by lowering PGN/Pam3CSK4 mediated nuclear cRel levels. Lower nuclear cRel level were observed to be because of enhanced transporting back rather than reduced nuclear translocation of cRel in MDP + PGN stimulated macrophages. These results demonstrate that Nod2 and TLR2/1 signaling pathways are independent and do not interact at the level of MAPK or NF-κB activation.     

侯晨曦：附表1-附表2

本研究基于7个叶绿体DNA片段（cpDNA）和2个核DNA片段（ITS和PZ8）的测序数据,对龙血树柴胡（Bupleurum dracaenoides Huan C.Wang,Z.R.He&H.Sun）8个居群的153个样本进行了遗传多样性和分布式样研究。cpDNA片段分析结果显示：龙血树柴胡在物种水平具有较高的遗传多样性（H_d=0.862;P_i=0.00567）,但居群内遗传多样性低,遗传变异主要存在于居群间,遗传分化显著（F_st=0.959）;而核DNA片段ITS和PZ8的数据分析结果显示,其遗传多样性较低（H_d=0.532,P_i =0.00121和H_d=0.349,P_i=0.00060）,遗传变异主要存在于居群内,居群间仅存在一定程度的遗传分化。中性检验和失配分布分析结果发现龙血树柴胡没有经历过近期种群扩张事件,8个居群的153个样本从遗传成份上可被分为两组。研究结果将为龙血树柴胡的资源保护和发掘提供参考。     

REGIOSELECTIVE 1-ALKYLATION OF 2′-DEOXYGUANOSINE

ABSTRACT

A method has been found for the regioselective alkylation of the nitrogen at the 1-position of 2′-deoxyguanosine. This consists in the reaction, in tetrahydrofuran solution, of a fully protected form of dG, namely the 3′5′-O-bis(tert-butyldimethylsilyl)-N ²-dimethylaminomethylene derivative, with an alkyl halide in the presence of cesium carbonate. The yields of these previously unavailable derivatives of 2′-deoxyguanosine range from good to excellent. Confirmation of the structure of these substances comes from a comparison of their spectroscopic properties with those of the known 1-methyl homologue. In particular, the UV spectra of these new derivatives and the known 1-methyl homologue are essentially identical.     

应用TaqMan定量方法研究巴马香猪CYP1A1、2A19、2E1 mRNA表达

目的巴马香猪是我国具有特色和优势的实验用小型猪资源品系,用于药物评价具有广阔前景。方法 以β-actin作校正,利用TaqMan定量技术对巴马香猪肝、肾、肾上腺、小肠、皮肤、脑、肺、睾丸、前列腺、子宫和卵巢等组织中CYP1A1、2A19和2E1 mRNA的表达水平进行检测,检测结果与报道的人体对应酶CYP1A2、2A6、2E1进行比较。结果巴马香猪CYP1A1、2A19、2E1 mRNA均以肝脏中最高,肝外组织明显较低,并且巴马香猪肝脏CYP1A1、2A19、2E1 mRNA均低于报道的人肝对应酶。结论巴马香猪CYP1A1、2A19、2E1与人体对应酶CYP1A2、2A6、2E1的mRNA组织表达存在一定差异,提示在其作为相应CYP亚型代谢的药物评价时应考虑这种种属差异对实验结果推广到人的影响。     

Action of phospholipase A2 of rabbit neuronal and glial cells on 1,2-diacyl-, 2-acyl-1-alk-1′-enyl-, and 2-acyl-1-alkyl-glycerophosphatides

Pronounced differences in the phospholipase A₂ activities were found in neurons and glia, the enzyme activity being two- to threefold higher in neurons than in glial cells. Both phospholipases A₂ hydrolyzed the 1,2-diacylglycerophosphatides more rapidly than the acylalkyl and acylalkenyl compounds. Choline plasmalogen and the corresponding alkyl derivative were cleaved at similar rates by the phospholipase A₂ from both glia and neurons. There was a tendency by the neuronal phospholipase A₂ to release arachidonic acid faster than linolenic acid from both phosphatidylcholine and ethanolamine, while arachidonic acid was removed less actively from phosphatidylethanolamine by the glial enzyme. The glial phospholipase A₂ showed a lag period of 10 or 20 min. Norepinephrine, injected into the lateral ventricle of the rabbit brain, stimulated the hydrolysis of the various 1,2-diacyl-, acylalkyl-, and acylalkenyl-glycerophosphatides by the phospholipase A₂ from both glia and neurons.