琼脂糖平板制备聚焦电泳法进一步提纯胸腺素组分五

用琼脂糖平板等电聚焦电泳法,由胸腺素组分五中分离出三种在聚焦电泳谱上是单一谱线的多肽成份——CP1、CP2和CP3,等电点分别为4.3、4.9和5.6。测定了这些多肽对脐带血中淋巴细胞形成羊红细胞玫瑰花的影响。与对照相比,CP1(2微克/0.6毫升),和CP3(0.2-2微克/毫升)分别在统计学上呈显著和非常显著差异。在相同测定条件下,这三种多肽成份的活性均高于化学合成的胸腺多肽——胸腺素α_1。     

Mechanism of erythromycin-induced ermC mRNA stability in Bacillus subtilis.

In Bacillus subtilis, the ermC gene encodes an mRNA that is unusually stable (40-min half-life) in the presence of erythromycin, an inducer of ermC gene expression. A requirement for this induced mRNA stability is a ribosome stalled in the ermC leader region. This property of ermC mRNA was used to study the decay of mRNA in B. subtilis. Using constructs in which the ribosome stall site was internal rather than at the 5' end of the message, we show that ribosome stalling provides stability to sequences downstream but not upstream of the ribosome stall site. Our results indicate that ermC mRNA is degraded by a ribonucleolytic activity that begins at the 5' end and degrades the message in a 5'-to-3' direction.     

胀果甘草化学成分的研究（Ⅲ）

继前文工作,本文报道从胀果甘草(Glycyrrhiza inflata Bat)根及根茎用95％乙醇渗滤后的提取部分中获得的另外五个化合物的结构鉴定。经理化性质及波谱分析,分别鉴定为胡萝卜甙(Daucosterol)、甘草查尔酮甲(Licochalcone A)、β-谷甾醇(β-Sitosterol),异芒柄花甙(Isoononin)和4＇,7一二羟基黄酮(4＇,7-Dihydroxy-flavone)。其中胡萝卜甙和异芒柄花甙为首次从该植物中获得。药理实验表明,甘草查尔酮甲对H_2O_2诱异的溶血有极好的抑制作用(97.3％),但甘草甙在三个体外氧化体系中都没有明显的活性。     

风速变化对草原生态系统的影响研究进展

在全球风速呈下降趋势的大背景下,研究风速变化对生态系统的影响具有重要意义,尤其是其重要组成部分——草原生态系统。近年来大量学者开始研究风速变化对草原生态系统的影响,主要集中在以下几个方面并得出相关的结论,(1)风速变化会影响植物的生长速率和叶片形态,适当的风速能够促进植物生长发育、提高植被初级生产力,而强风或持续大风不仅会对植物产生破坏作用,还会影响其生长发育;(2)风会最先带走地表细小颗粒,从而导致土壤质地变粗、水分下降、营养成分重新分配;(3)风引起地表边界层和大气边界层物质和能量的转移和交换,热量和水汽的交换导致地表微气候发生变化,如风速降低会导致地表温度升高;(4)风力作用使得土壤水分亏缺、营养成分变化,导致草原生态系统结构变化、草地覆盖度降低、物种生活型复杂化、耐旱植物增加;(5)大气稳定性、CO2交换速率和碳排放都会随着风速的增加而增加,碳吸收则相反,碳通量也因此发生变化。综上,风速降低对于草原生态系统的影响复杂且利弊相当,未来的发展趋势会更加侧重于以下几个方面的发展:研究对象的多样化、加强控制实验的定量化研究、综合多要素的相互作用机理研究、整体结构和功能性的研究。     

Site-directed mutagenesis and functional analysis of active site acidic amino acid residues D142, D144 and E146 in Manduca sexta (tobacco hornworm) chitinase

Chitinases (EC 3.2.1.14) are glycosyl hydrolases that catalyze the hydrolysis of beta-(1, 4)-glycosidic bonds in chitin, the major structural polysaccharide present in the cuticle and gut peritrophic matrix of insects. Two conserved regions have been identified from amino acid sequence comparisons of family 18 glycosyl hydrolases, which includes Manduca sexta (tobacco hornworm) chitinase as a member. The second of these regions in M. sexta chitinase contains three very highly conserved acidic amino acid residues, D142, D144 and E146, that are probably active site residues. In this study the functional roles of these three residues were investigated using site-directed mutagenesis for their substitutions to other amino acids. Six mutant proteins, D142E, D142N, D144E, D144N, E146D and E146Q, as well as the wild-type enzyme, were produced using a baculovirus-insect cell line expression system. The proteins were purified by anion-exchange chromatography, after which their physical, kinetic and substrate binding properties were determined. Circular dichroism spectra of the mutant proteins were similar to that of the wild-type protein, indicating that the presence of mutations did not change the overall secondary structures. E146 was required for enzymatic activity because mutants E146Q and E146D were devoid of activity. D144E retained most of the enzymatic activity, but D144N lost nearly 90%. There was a shift in the pH optimum from alkaline pH to acidic pH for mutants D142N and D144E with minimal losses of activity relative to the wild-type enzyme. The pH-activity profile for the D142E mutation resembled that of the wild-type enzyme except activity in the neutral and acidic range was lower. All of the mutant proteins bound to chitin. Therefore, none of these acidic residues was essential for substrate binding. The results indicate that E146 probably functions as an acid/base catalyst in the hydrolytic mechanism, as do homologous residues in other glycosyl hydrolases. D144 apparently functions as an electrostatic stabilizer of the positively charged transition state, whereas D142 probably influences the pKa values of D144 and E146.     

MicroRNA-30a sensitizes tumor cells to cis-platinum via suppressing beclin 1-mediated autophagy

Autophagy is activated in cancer cells during chemotherapy and often contributes to tumor chemotherapy resistance. In this study, we characterized the role of microRNA-30a (miR-30a) in the coordination of cancer cell apoptosis and autophagy, which determines the sensitivity of cancer cells to chemotherapy. First, the autophagy activity in cancer cells increased after cis-dichloro-diamine platinum (cis-DDP) or Taxol treatment, as indicated by the enhanced expression of beclin 1, a key regulator of autophagy, and increased number of LC3-positive autophagosomes. Second, miRNA screening using a TaqMan probe-based quantitative RT-PCR assay identified that miR-30a, a miRNA that targets beclin 1, was significantly reduced in tumor cells by cis-DDP treatment. Forced expression of miR-30a significantly reduced beclin 1 and the autophagy activity of tumor cells induced by cis-DDP. Third, the blockade of tumor cell autophagy activity by miR-30a expression or 3-methyladenine significantly increased tumor cell apoptosis induced by cis-DDP treatment. Finally, an in vivo tumor implantation mouse model clearly showed that elevation of miR-30a in implanted tumor cells by administration of the recombinant lentivirus expressing miR-30a strongly enhanced cis-DDP-induced apoptosis of tumor cells. In conclusion, our results demonstrate for the first time that miR-30a can sensitize tumor cells to cis-DDP via reducing beclin 1-mediated autophagy and that increasing miR-30a level in tumor cells represents a novel approach to enhance the efficacy of chemotherapy during cancer treatment.     

Screening and identification of extracellular lipase-producing thermophilic bacteria from a Malaysian hot spring

Seven lipase-producing thermophilic bacteria (ST 1, ST 4, ST 6, ST 7, ST 8, ST 9 and ST 10) were isolated from the Setapak hot spring in Malaysia. The crude extracellular lipases recovered by ultrafiltration of cell-free culture supernatant were reacted in an olive oil mixture and their lipolytic activities were compared. Identification of the bacteria was carried out using the Biolog system and biochemical tests. Strain ST 7 that exhibited the highest lipolytic activity of 4.58 U/ml was identified as belonging to the Bacillus genus. Strain ST 6 with an activity of 3.51 U/ml, was identified as Ralstonia paucula. The lipolytic activities of strains ST 1, ST 4, ST 8, ST 9 and ST 10 were 2.39, 1.84, 2.38, 1.80 and 2.62 U/ml respectively. Strains ST 1, ST 4, and ST 10 were identified as Ralstonia paucula while strains ST 8 and ST 9 were Bacillus spp. Strains ST 7 and ST 9 were tentatively identified as Bacillus thermoglucosidasius, Bacillus stearothermophilus or Bacillus coagulans, whereas strain ST 8 was tentatively identified as Bacillus subtilis.     

LncRNA MEG3 inhibits the progression of prostate cancer by modulating miR‐9‐5p/QKI‐5 axis

This study was designed to detecting the influences of lncRNA MEG3 in prostate cancer. Aberrant lncRNAs expression profiles of prostate cancer were screened by microarray analysis. The qRT‐PCR and Western blot were employed to investigating the expression levels of lncRNA MEG3, miR‐9‐5p and QKI‐5. The luciferase reporter assay was utilized to testifying the interactions relationship among these molecules. Applying CCK‐8 assay, wound healing assay, transwell assay and flow cytometry in turn, the cell proliferation, migration and invasion abilities as well as apoptosis were measured respectively. LncRNA MEG3 was a down‐regulated lncRNA in prostate cancer tissues and cells and could inhibit the expression of miR‐9‐5p, whereas miR‐9‐5p down‐regulated QKI‐5 expression. Overexpressed MEG3 and QKI‐5 could decrease the abilities of proliferation, migration and invasion in prostate cancer cells effectively and increased the apoptosis rate. On the contrary, miR‐9‐5p mimics presented an opposite tendency in prostate cancer cells. Furthermore, MEG3 inhibited tumour growth and up‐regulated expression of QKI‐5 in vivo. LncRNA MEG3 was a down‐regulated lncRNA in prostate cancer and impacted the abilities of cell proliferation, migration and invasion, and cell apoptosis rate, this regulation relied on regulating miR‐9‐5p and its targeting gene QKI‐5.     

三维技术对安徽白鹭洲战国墓M566墓主容貌的复原

2011年安徽白鹭洲发现战国时期保存完整的楚国贵族墓葬,因墓主人头发和发簪保存完好、墓主身份显赫而备受关注。该头骨保存完整,具有亚洲蒙古人种的特征,根据骨骼的形态推测其为女性,年龄为35-39岁。为展现该地区战国贵族妇女的容貌、丰富该地区考古多样性提供研究材料。本文首先使用高分辨率CT对该个体的头骨及下颌骨进行了扫描和重建,然后采用基于偏最小二乘回归的颅面复原方法实现生前容貌基本形态的复原。最后,结合考古资料,利用三维模型处理软件对面貌复原模型及其发饰进行了三维建模和纹理贴图等处理,提高了颅面复原模型的真实感,生动形象地再现战国贵族女性面部的形态特征。