P63和TTF-1在肺癌组织中的表达及意义

目的研究P63和TTF-1在肺癌各种类型组织中的表达及其意义。方法随机收集原发性肺癌组织标本53例（其中鳞癌16例,腺癌16例,小细胞癌14例,大细胞癌7例,均为中低分化程度癌组织）,采用免疫组织化学S-P法分别检测P63蛋白和TTF-1蛋白在各种类型肺癌组织中的表达并结合二者表达的结果进行分析。结果二者的表达在肺癌细胞中定位于细胞核,呈棕黄色。在肺鳞癌、肺腺癌、肺小细胞癌和肺大细胞癌中,P63的阳性表达率分别为93.8%（15/16）、37.5%（6/16）、21.4%（3/14）、28.6%（2/7）;TTF-1的阳性表达率分别为18.8%（3/16）、75%（12/16）、78.6%（11/14）、0%（0/7）。结论①P63在肺鳞癌中的表达水平较高,可以作为鉴别低分化鳞癌与低分化腺癌,小细胞癌的指标;②TTF-1在低分化腺癌和小细胞癌中的表达水平较高,对于肺癌组织类型和非癌组织的鉴别具有一定意义;③根据P63和TTF-1在肺癌组织的特异性表达,将二者联合起来有助于对低分化鳞癌和低分化腺癌以及低分化鳞癌和小细胞癌的鉴别诊断。     

Intragenic spatial patterns of codon usage bias in prokaryotic and eukaryotic genomes

To study the roles of translational accuracy, translational efficiency, and the Hill-Robertson effect in codon usage bias, we studied the intragenic spatial distribution of synonymous codon usage bias in four prokaryotic (Escherichia coli, Bacillus subtilis, Sulfolobus tokodaii, and Thermotoga maritima) and two eukaryotic (Saccharomyces cerevisiae and Drosophila melanogaster) genomes. We generated supersequences at each codon position across genes in a genome and computed the overall bias at each codon position. By quantitatively evaluating the trend of spatial patterns using isotonic regression, we show that in yeast and prokaryotic genomes, codon usage bias increases along translational direction, which is consistent with purifying selection against nonsense errors. Fruit fly genes show a nearly symmetric M-shaped spatial pattern of codon usage bias, with less bias in the middle and both ends. The low codon usage bias in the middle region is best explained by interference (the Hill-Robertson effect) between selections at different codon positions. In both yeast and fruit fly, spatial patterns of codon usage bias are characteristically different from patterns of GC-content variations. Effect of expression level on the strength of codon usage bias is more conspicuous than its effect on the shape of the spatial distribution.     

Preliminary results on nematicidal activity from culture filtrates of Basidiomycetes against the pine wood nematode,Bursaphelenchus xylophilus (Aphelenchoididae)

One hundred and eighty one fungal species that were isolated from the fresh fruiting bodies collected in the Mountains of Pu Er County of Yunnan Province, China were tested on the pine wood nematode,Bursaphelenchus xylophilus in vitro. Each fungal species was grown in Czapek broth and potato dextrose broth (PDB). Fifteen filtrates fromAmauroderma austrosinense, Amauroderma macer, Filoboletus sp.,Laccaria tortilis, Lactarius gerardii, Lentinula edodes, Oudemansiella longipes, Oudemansiella mucida, Peziza sp.,Pleurotus sp.,Sinotermitomyces carnosus (two strains),Strobilomyces floccopus, Termitomyces albuminosus, Tylopilus striatulus grown on PDB were found to be pathogenic to the tested nematodes. Eleven filtrates fromAmanita junguillea, Amanita sp.,Daedalea sepiaria, Fistulina hepatica, Omphalotus olearius, Oudemansiella mucida, Peziza sp.,Pleurotus pulmatus, Ramaria sp.,Tricholoma conglobatum, Tylopilus striatulus grown on Czapek broth were also pathogenic to the nematodes. When screening for nematicidal potential of fungi, it is important to study the growth medium conditions necessary to obtain the optimal nematicidal effect as fungal filtrates growing on different liquid media showed a very inconsistent toxicity towards nematodes.     

Alpha-melanocyte stimulating hormone protects retinal pigment epithelium cells from oxidative stress through activation of melanocortin 1 receptor–Akt–mTOR signaling

Patients with age related macular degeneration (AMD) will develop vision loss in the center of the visual field. Reactive oxygen species (ROS)-mediated retinal pigment epithelium (RPE) cell apoptosis is an important contributor of AMD. In this study, we explored the pro-survival effect of α-melanocyte stimulating hormone (α-MSH) on oxidative stressed RPE cells. We found that α-MSH receptor melanocortin 1 receptor (MC1R) was functionally expressed in primary and transformed RPE cells. RPE cells were response to α-MSH stimulation. α-MSH activated Akt/mammalian target of rapamycin (mTOR) and Erk1/2 signalings in RPE cells, which were inhibited by MC1R siRNA knockdown. α-MSH protected RPE cells from hydrogen peroxide (H₂O₂)-induced apoptosis, an effect that was almost abolished when MC1R was depleted by siRNA. α-MSH-mediated S6K1 activation and pro-survival effect against H₂O₂ was inhibited by Akt inhibitors (perifosine, MK-2206 and LY294002). Further, mTOR inhibition by rapamycin, or by mTOR siRNA knockdown, diminished α-MSH’s pro-survival effect in RPE cells. Thus, Akt and its downstream mTOR signaling mediates α-MSH-induced survival in RPE cells. In summary, we have identified a new α-MSH–MC1R physiologic pathway that reduces H₂O₂-induced RPE cell damage, and might minimize the risk of developing AMD.     

菠菜BADH单抗细胞株及其应用初探

用菠菜甜菜碱醛脱氢酶 ( BADH)免疫巴比西 ( BALB/c)小鼠 ,将其脾细胞与骨髓瘤细胞 SP2 /O-Ag1 4融合 ,在 1 92孔中 ,有约 1 4 %孔生长的杂交瘤细胞 ,用间接酶联免疫方法 ( ELISA)检测表现为阳性。选择其中 2 G3和 2 D10 细胞系 ,用有限稀释法进行克隆化培养 ,约 2 0 %克隆化细胞为强阳性。选择其中 2 G3- H3细胞株注射到 BALB/c小鼠腹腔中诱导腹水 ,腹水的单抗效价为 1∶ 1 0 3。应用 BADH单抗检查了大麦、水稻、高粱、小麦幼苗的叶片和根的粗提物 ,均呈阳性反应 ,表明 BADH除在光合组织中存在外 ,在非光合组织中也可能存在。讨论了非光合组织 BADH的意义     

休眠期和营养期包囊游仆虫的纤毛器骨架及其微管蛋白

应用光镜和透射电镜术 ,显示了包囊游仆虫休眠细胞中纤毛器骨架的形态 ,并对该纤毛虫休眠细胞和营养细胞的纤毛器及其α、β -微管蛋白进行了免疫荧光定位的比较研究。由免疫荧光显微术显示 ,包囊游仆虫形成休眠包囊后 ,背部毛基体完整地按原有模式保存下来 ;纤毛杆解聚后微管蛋白多集中在细胞皮层 ,小部分均匀散布在细胞质中。据所得结果认为 ,包囊游仆虫形成包囊后 ,微管蛋白主要有 3个去向 ,即 :①处于自噬泡内被逐步消化 ;②以“微管蛋白库”的形式分布于细胞皮层及细胞质中 ;③保留在残留的基体中。此外 ,以往研究中发现的棘毛基部纤维网络未被标记上 ,提示这些纤维体系可能不属于微管系统     

树载脂蛋白CIcDNA序列及其组织表达

分离提取树（ｔｒｅｅｓｈｒｅｗ,ＴＳ）肝组织ｍＲＮＡ,以此为模板,反转录构建了ＴＳ肝组织ｃＤＮＡ文库。利用制备的兔抗ＴＳ载脂蛋白ＣＩ多抗血清为探针筛选该文库,获得两个阳性克隆。测序及序列分析表明：两个克隆均为ＴＳａｐｏＣＩｃＤＮＡ基因。大的克隆由３８０个核苷酸构成,包括２１ｂｐ、９５ｂｐ组成的５′和３′非编码区,２６４ｂｐ组成的一个完整开放阅读框架,编码８８个氨基酸的ａｐｏＣＩ前体,含２６个氨基酸构成的信号肽和６２肽的成熟蛋白。其成熟肽长度与大鼠、小鼠和狗的相应结构一致,但较人和狒狒ａｐｏＣＩ的成熟肽多５个氨基酸。并对其功能域进行了初步预测。ＲＮＡ印迹显示ＴＳａｐｏＣＩｍＲＮＡ不仅主要在肝组织表达;而且不同于人和其他哺乳类动物,亦可少量在小肠表达。上述结果为进一步研究ＴＳａｐｏＣＩ基因组结构、功能和体外表达奠定了基础。     

BMP2/7 heterodimer enhances osteogenic differentiation of rat BMSCs via ERK signaling compared with respective homodimers

Bone morphogenetic protein (BMP)2/7 heterodimer shows greater efficacy in enhancing bone regeneration. However, the precise mechanism and the role of mitogen-activated protein kinase (MAPK) signaling network in BMP2/7-driven osteogenesis remain ambiguous. In this study, we evaluated the effects of BMP2/7 heterodimers on osteoblastic differentiation in rat bone marrow mesenchymal stem cells (BMSCs), with the aim to elaborate how MAPKs might be involved in this cellular process by treatment of rat BMSCs with BMP2/-7 with a special signal-pathway inhibitor. We found that BMP2/7 heterodimer induced a much stronger osteogenic response in rat BMSCs compared with either homodimer. Most interestingly, extracellular signal-regulated kinase (ERK) demonstrated a highly sustained phosphorylation and activation in the BMP2/7 heterodimer treatment groups, and inhibition of ERK cascades using U0126 special inhibitor that significantly reduced the activity of ALP and calcium mineralization to a substantial degree in rat BMSCs treated with BMP2/7 heterodimers. Collectively, we demonstrate that BMP2/7 heterodimer shows a potent ability to stimulate osteogenesis in rat BMSCs. The activated ERK signaling pathway involved in this process may contribute partially to an increased osteogenic potency of heterodimeric BMP2/7 growth factors.     

ELK-1/JNK/c-Fos对左归降糖解郁方(ZGJTJYF)体外培养的模拟糖尿病并发抑郁症(DD)大鼠海马神经元凋亡中的作用

目的:研究信号通路ELK-1/JNK/c-Fos在左归降糖解郁方(ZGJTJYF)对模拟糖尿病并发抑郁症(DD)环境下抗海马神经元凋亡中的作用。方法:原代培养海马神经元,加入高糖(150 mmol/L)+皮质酮(200μmol/L),构建DD体外细胞模型;将培养的海马神经元细胞随机分为5组:空白血清组、正常组、左归降糖解郁方含药血清组、阳性药(二甲双胍+氟西汀)含药血清组和模型组(每组3个复孔)。模型组和正常组给予等量培养液,其余组加入相应体积分数10%的相应血清,均干预18 h。分别采用Hoechst染色、高内涵细胞成像分析技术和RT-PCR技术分别检测海马神经元凋亡情况及检测凋亡相关ELK-1、JNK和c-Fos蛋白和基因的表达。结果:与空白组比较,模型组细胞凋亡明显,其海马神经元凋亡数量明显增多,ELK-1、JNK和c-Fos的mRNA及蛋白表达水平均显著升高(P<0.05);与模型组比较,左归降糖解郁方含药血清组和阳性药含药血清组大鼠海马神经元可见局部亮点明显减少,凋亡细胞数显著减少,ELK-1、JNK和c-Fos蛋白及mRNA表达均明显下调(P<0.05),且神经网络及树突连接情况得到明显改善。结论:左归降糖解郁方可以减低DD环境下海马神经元中Elk-1、JNK、c-fos表达而起到抗凋亡的效果。