Identification of microRNAs in Wool Follicles during Anagen,Catagen, and Telogen Phases in Tibetan Sheep

Background

Wool quality is one of the most important economic traits in sheep. The wool fiber is derived from specialized skin cells that are referred to as wool follicles. To understand the roles of microRNAs (miRNAs) in wool fiber growth, we detected the expression patterns of miRNAs in wool follicles at the anagen, catagen, and telogen stages from Tibetan sheep through Solexa sequencing.

Results

A total of 244 mature miRNAs were identified. Of these, only five miRNAs are listed in the database of sheep miRNAs (miRBase Database V19), and the other 239 miRNAs have not been previously described in this species. Further analyses indicated that 204 miRNAs are evolutionarily conserved among mammal species, whereas 35 of the identified miRNAs were first found specifically in sheep. The expression pattern analyses showed that the expression levels of 39, 34, and 20 of the miRNAs significantly change between anagen and catagen, between anagen and telogen, and between catagen and telogen, respectively. The results of the bioinformatics analysis show that these differentially expressed miRNAs might regulate wool follicle development by targeting genes in many different pathways, such as the MAPK and Wnt pathways, as well as the pathways that regulate the actin cytoskeleton, focal adhesion, and tight junctions. Furthermore, we identified six differentially expressed miRNAs (oar-miR-103-3P, oar-miR-148b-3P, oar-miR-320-3P, oar-miR-31-5P, oar-novel-1-5P, and oar-novel-2-3P) that might target the key genes of the Wnt pathway. It has been reported that the Wnt pathway is critical for wool follicle development. Therefore, these miRNAs may regulate wool development through the Wnt pathway.

Conclusions

Our results provide new information on the identification and expression pattern of miRNAs in wool follicles. Our data might therefore aid in the understanding of the mechanisms of wool follicle development in sheep.     

东北红豆杉(Taxus cuspidata)中四种紫杉烷类化合物抑制妇科肿瘤细胞增殖活性研究及其作用机制的初步探讨

通过四唑盐(MTT)比色法检测东北红豆杉中四种紫杉烷类化合物对人子宫颈癌细胞、人子宫内膜癌细胞、人卵巢透明癌细胞和人卵巢囊腺癌细胞增殖的影响,不仅探讨了这些化合物作用的构效关系,还初步探讨了其抑制肿瘤细胞增殖的作用机制。结果发现9,10-去乙酰紫杉宁对人子宫颈癌细胞、人子宫内膜癌细胞和人卵巢囊腺癌细胞的增殖均有明显的抑制作用,与紫杉宁和1β-羟基紫杉宁即使在100μmol/L的高浓度下,对五种妇科肿瘤细胞的增殖不显示抑制活性;流式细胞学检查发现9,10-去乙酰紫杉宁作用后的HeLa细胞出现凋亡现象。因此推测:1.C-9,10位的羟基和C-5位的肉桂酰基侧链是该类化合物抑制肿瘤细胞增殖所必需的;2.9,10-去乙酰紫杉宁有可能是通过诱导肿瘤细胞凋亡来实现其抑制HeLa细胞的增殖作用。     

HCV编码病毒结构蛋白的DNA免疫研究

应用HCVC (pC)和HCVCE1E2 (pCE1E2 )重组体转染真核细胞并且通过肌注免疫BALB/C小鼠 ,对其体液免疫和细胞免疫进行检测。所用的 pC和 pCE1E2 均可在真核细胞内表达出特异性HCVC蛋白 ;肌注DNA免疫后均可诱导出BALB/C小鼠的体液和细胞免疫反应 ,抗体反应的A值 :pC组为 0 .358± 0 .0 96 ,pCE1E2 组为 0 .4 15± 0 .12 7;CTL活力 pC组为 18.6 5%± 5.72 % ,pCE1E2 组为 2 0 .0 7%± 11.11% ;通过免疫鼠荷瘤检测体内CTL反应 ,可观察到免疫组鼠发瘤时间滞后 ,发瘤部位减少和存活时间延长。在开发和研制HCV疫苗的过程中 ,DNA免疫是在快速构建、评价和筛选免疫原方面的有效办法。     

1.42A crystal structure of mini-IGF-1(2): an analysis of the disulfide isomerization property and receptor binding property of IGF-1 based on the three-dimensional structure

Insulin and insulin-like growth factor 1 (IGF-1) share a homologous sequence, a similar three-dimensional structure and weakly overlapping biological activity, but IGF-1 folds into two thermodynamically stable disulfide isomers, while insulin folds into one unique stable tertiary structure. This is a very interesting phenomenon in which one amino acid sequence encodes two three-dimensional structures, and its molecular mechanism has remained unclear for a long time. In this study, the crystal structure of mini-IGF-1(2), a disulfide isomer of an artificial analog of IGF-1, was solved by the SAD/SIRAS method using our in-house X-ray source. Evidence was found in the structure showing that the intra-A-chain/domain disulfide bond of some molecules was broken; thus, it was proposed that disulfide isomerization begins with the breakdown of this disulfide bond. Furthermore, based on the structural comparison of IGF-1 and insulin, a new assumption was made that in insulin the several hydrogen bonds formed between the N-terminal region of the B-chain and the intra-A-chain disulfide region of the A-chain are the main reason for the stability of the intra-A-chain disulfide bond and for the prevention of disulfide isomerization, while Phe B1 and His B5 are very important for the formation of these hydrogen bonds. Moreover, the receptor binding property of IGF-1 was analyzed in detail based on the structural comparison of mini-IGF-1(2), native IGF-1, and small mini-IGF-1.     

日本血吸虫虫卵、童虫和雌雄成虫膜蛋白的双向电泳

血吸虫寄生生活复杂,中间宿主和终末宿主转换,有性繁殖和无性繁殖交替,由其感染引起的血吸虫病仍严重危害人畜健康(陈贤义等,2002).研究表明,血吸虫不仅能利用宿主的免疫信号分子伪装自己获得宿主的免疫兼容(Salzet et al.,2000),而且还在血吸虫中克隆到宿主信号分子的受体(Ahmed et al.,2001).     

Wistar大鼠实验性变态反应性脑脊髓炎病理探讨

目的研究缓解-复发型EAE大鼠的基本病理改变。方法对缓解-复发型EAE大鼠进行HE染色、Weil髓鞘染色和改良的Bielschowsky染色,以免疫组化方法标记GFAP和MMP-2、MMP-9。结果组织内可见多个血管周围淋巴细胞浸润呈袖套样分布,伴有大片状脱髓鞘,部分脱失的髓鞘内有炎细胞浸润,轴索呈空泡样缺失,GFAP染色证明在旧病灶的周围部分可见星形胶质细胞增生,MMP-2、MMP-9在血管内皮细胞、细胞外基质、炎细胞及脑(脊)膜内呈阳性表达。结论①缓解-复发型EAE大鼠的组织病理学改变与临床表现一致,有活动性和非活动性病灶并存的现象。②MMP-2、MMP-9在活动性病灶内表达,参与疾病的发生。③缓解-复发型EAE病理改变与MS相似,是理想的动物模型。     

Co-expression of P450 BM3 and glucose dehydrogenase by recombinant <Emphasis Type="Italic">Escherichia coli</Emphasis> and its application in an NADPH-dependent indigo production system

P450 BM3 mutant can catalyze indole to indoxyl, and indoxyl can dimerize to form indigo. But the reaction catalyzed by P450 BM3 requires NADPH, as coenzyme regeneration is very important in this system. As we know, when glucose dehydrogenase oxidizes glucose to glucolactone, NADH or NADPH can be formed, which can contribute to NADPH regeneration in the reaction catalyzed by P450 BM3. In this paper, a recombinant Escherichia coli BL21 (pET28a (+)-P450 BM3-gdh0310) was constructed to co-express both P450 BM3 gene and glucose dehydrogenase (GDH) gene. To improve the expression level of P450 BM3 and GDH in E. coli and to avoid the complex and low-efficiency refolding operation in the purification procedure, the expression conditions were optimized. Under the optimized conditions, the maximum P450 BM3 and GDH activities amounted to 8173.13 and 0.045 U/mg protein, respectively. Then bioconversion of indole to indigo was carried out by adding indole and glucose to the culture after improved expression level was obtained under optimized conditions, and 2.9 mM (760.6 mg/L) indigo was formed with an initial indole concentration of 5 mM.     

Multiple effects of differentiation-inducing factor on prespore differentiation and cyclic-AMP signal transduction in Dictyostelium

Abstract. The effects of the differentiation inducing factor (DIF) on several CAMP-induced responses in Dictyostelium were investigated. It was found that DIF reduces the apparent affinity of cell-surface cAMP receptors. DIF does not affect the CAMP-induced cGMP response, but it is a potent inhibitor of the CAMP-relay response. DIF also inhibits the induction of prespore differentiation by cAMP in aggregation-competent cells. We also compared the effects of DIF on CAMP-induced responses with those of the relay inhibitor, caffeine, and the morphogen, adenosine.     

外源氮输入和水分变化对荒漠草原凋落物分解的影响

全球气候变化背景下,大气氮沉降和降水变化日益显著,其对荒漠草原凋落物分解的影响存在很大的不确定性.采用裂区设计,设置主区为自然降雨、增雨30%和减雨30% 3个水分处理,副区为0(N₀)、30(N₃₀)、50(N₅₀)和100 kg·hm^-2·a^-1(N₁₀₀)4个氮素水平,经过21个月(2016年1月—2017年10月)水氮处理,研究水氮共同作用对荒漠草原常见物种猪毛菜、短花针茅和木地肤3种植物凋落物分解的影响.结果表明: 3种凋落物干物质残留率随时间增加而减少,用Olson负指数衰减模型拟合效果较好,凋落物分解系数(k)大小为猪毛菜>短花针茅>木地肤.增雨30%N₁₀₀处理分解系数最高,为0.028.单因素处理下,增雨30%和N₅₀的凋落物分解最快.水氮共同作用下,增雨 30%N₁₀₀处理凋落物分解最快.3种凋落物初始化学全氮含量大小为猪毛菜>短花针茅>木地肤,猪毛菜和短花针茅k值与全氮含量呈显著正相关;全碳含量、纤维素含量、木质素含量、C/N、木质素/N和纤维素/N大小为木地肤>短花针茅>猪毛菜,猪毛菜k值与各指标均呈显著负相关,短花针茅和木地肤k值与C/N、木质素/N和纤维素/N均呈显著负相关.猪毛菜分解最快,木地肤分解最慢.适量的水、氮添加有利于荒漠草原凋落物的分解,可以促进土壤养分循环,对荒漠草原可持续发展及生态平衡有积极作用.