Inducing dopaminergic differentiation of expanded rat mesencephalic neural stem cells by ascorbic acid in vitro

Ascorbic acid (AA) induced differentiation of neural stem cells (NSCs) into dopaminergic (DAergic) neurons is reported.NSCs derived from rat mesencephalon were maintained and expanded in a defined medium containing mitogens of basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF).Compared with the control, ascorbic acid treatment led to more DAergic neuronal differentiation as indicated by the expression of tyrosine hydroxylase (TH) and dopamine transporter (DAT), which are specific markers of dopamine neurons.AA induction also enhanced expression of Nurr1 and Shh.PD98059, an inhibitor of mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) pathway, could block AA-induced Nurr1, TH and DAT mRNA expression.The results might suggest a new strategy to provide enough dopaminergic cells for the therapy of Parkinson's disease (PD), and Nurr1 and ERK signaling pathway might participate in the AA-induced DAergic differentiation.     

酿酒酵母蛋白酶A外泌与酒类酿造

蛋白酶A(EC 3.4.23.6)是酿酒酵母中重要的蛋白酶,与酶原激活、生孢等多种生理功能密切相关.在酒类酿造过程中往往会发生蛋白酶A外泌进而影响发酵产品质量的现象.研究胁迫条件下蛋白酶A的外泌及其调控机制,对于酿酒酵母菌种选育和酒类产品质量控制具有重要的理论指导意义.本文主要对蛋白酶A外泌机制和蛋白酶A外泌对酒类酿造影响的相关研究进行综述.     

Salicylic acid and ethylene signaling pathways are involved in production of rice trypsin proteinase inhibitors induced by the leaf folder Cnaphalocrocis medinalis （Guenee）

The roles of signaling pathways in the production of trypsin proteinase inhibitors (TrypPIs) in rice infested by the leaf folder (LF) Cnaphalocrocis medinalis were studied. Infestation by LF increased TrypPI levels in the leaves of rice plants at the tillering, booting and flowering stages but decreased TrypPI levels at the ripening stage; TrypPI levels in rice stems did not increase at any developmental stage. Infestation by LF at the tillering stage systemically increased TrypPI levels in leaves but not in stems; it also enhanced salicylic acid (SA) levels in leaves and stems, and the ethylene level released from plants. However, LF infestation did not increase JA concentrations. Exogenous application of SA or ethylene enhanced TrypPI levels in the leaves and stems of plants at the tillering stage, whereas treatment with both SA and ethylene induced lower levels of TrypPIs than treatment with SA or ethylene alone, suggesting an antagonistic effect of SA and ethylene on TrypPIs induction. The results suggest that both SA and ethylene signaling pathways are involved in the production of TrypPIs in rice induced by LF; moreover, the antagonistic effect of SA and ethylene may explain the changes in TrypPI levels seen at different plant developmental stages and in different organs.     

水稻胰蛋白酶抑制剂对褐飞虱产卵刺激的响应

 昆虫产卵过程对寄主植物的直接刺激包括产卵器的机械损伤和卵表面物质带来影响,这些刺激是启动植物诱导抗虫性的重要途径。通过对褐飞虱产卵处理后水稻植株内胰蛋白酶抑制剂（BBPI）合成酶基因的表达量及物质含量的测定,对比褐飞虱取食和机械损伤处理,发现褐飞虱产卵能够诱导水稻BBPI合成酶基因的表达和BBPI物质的生成。处理后6 h,12 h,水稻 BBPI 表达量显著高于取食和机械损伤处理;产卵诱导的BBPI物质含量显著高于正常水稻。褐飞虱产卵诱导水稻启动〖WTBX〗BBPI〖WTBZ〗表达和物质合成表明,与取食危害类似,水稻同样会对褐飞虱产卵刺激产生响应,启动相应的植物防御体系,进而达到阻滞褐飞虱危害的目的。     

In vivo matrix-guided human mesenchymal stem cells

Microfracture of subchondral bone results in intrinsic repair of cartilage defects. Stem or progenitor cells from bone marrow have been proposed to be involved in this regenerative process. Here, we demonstrate for the first time that mesenchymal stem (MS) cells can in fact be recovered from matrix material saturated with cells from bone marrow after microfracture. This also introduces a new technique for MS cell isolation during arthroscopic treatment. MS cells were phenotyped using specific cell surface antibodies. Differentiation of the MS cells into the adipogenic, chondrogenic and osteogenic lineage could be demonstrated by cultivation of MS cells as a monolayer, as micromass bodies or mesenchymal microspheres. This study demonstrates that MS cells can be attracted to a cartilage defect by guidance of a collagenous matrix after perforating subchondral bone. Protocols for application of MS cells in restoration of cartilage tissue include an initial invasive biopsy to obtain the MS cells and time-wasting in vitro proliferation and possibly differentiation of the cells before implantation. The new technique already includes attraction of MS cells to sites of cartilage defects and therefore may overcome the necessity of in vitro proliferation and differentiation of MS cells prior to transplantation. Received 3 November 2005; received after revision 15 December 2005; accepted 4 January 2006     

鲨鱼肠道蛋白酶的分离纯化及性质的初步研究

以鲨鱼肠为材料，经柠檬酸缓冲液抽提，硫酸铵分级分离，ＣＭ－纤维素离子交换柱层析纯化，获得蛋白酶（Ⅰ）和蛋白酶（Ⅱ），研究了蛋白酶Ⅱ性质，酶经聚丙烯酰胺凝胶电泳鉴定为单一蛋白纯，酶的紫外吸收特征峰在２７５ｎｍ处，测得酶的分子量为１８ｋｄ，在ＰＨ８．０、３７℃下酶催化酪蛋白水解的Ｋｍ值为６．９×１０＾－５ｍｏｌ／Ｌ。酶水解酪蛋白的最活温度为５２℃，活化能为７６．５３ｋＪ／ｍｏｌ。几种金属离子对酶活力的     

鲨鱼肠蛋白酶活性必需基团的研究

用化学修饰法结合酶的紫外吸收光谱变化研究灰星鲨（ＭｕｓｔｅｔｕｓＧｒｉｓｅｕｓ）肠蛋白酶的功能基团性质，结果表明：色氨酸残基、组氨酸残基、赖氨酸残基及精氨酸残基均与酶活性有关，而羧基、巯基与酶活性无关。     

菠萝蛋白酶的基本性质研究

本文介绍了菠萝蛋白酶的紫外吸收光谱、分子量、米氏常数 K_■、激活剂等抟影响,以及最适的 pH 和温度、热失活情况等基本性质.实验表明菠萝蛋白酶为非金属酶;发现不同来源的菠萝蛋白酶的最适 pH 为中性,都具有一定的耐热性.     

Profiling of the secreted proteins during 3T3-L1 adipocyte differentiation leads to the identification of novel adipokines

Adipose tissue is an endocrine organ capable of secreting a number of adipokines with a role in the regulation of adipose tissue and whole-body metabolism. We used two-dimensional gel electrophoresis combined with mass spectrometry to profile the secreted proteins from (pre)adipocytes. The culture medium of 3T3-L1 cells during adipocyte differentiation was screened, and 41 proteins that responded to blocking of secretion by 20°C treatment and/or brefeldin A treatment were identified. Prohibitin, stress-70 protein, and adhesion-regulating molecule 1 are reported for the first time as secreted proteins. In addition, procollagen C-proteinase enhancer protein, galectin-1, cyclophilin A and C, and SF20/IL-25 are newly identified as adipocyte secreted factors. Secretion profiles indicated a dynamic environment including an actively remodeling extracellular matrix and several factors involved in growth regulation.Received 15 June 2004; received after revision 26 July 2004; accepted 2 August 2004     

Specific interactions of the adenovirus proteinase with the viral DNA,an 11-amino-acid viral peptide,and the cellular protein actin

The adenovirus proteinase (AVP) is synthesized in an inactive form that requires cofactors for activation. The interaction of AVP with two viral cofactors and with a cellular cofactor, actin, is characterized by quantitative analyses. The results are consistent with a specific model for the regulation of AVP. Late in adenovirus infection, inside nascent virions, AVP becomes partially activated by binding to the viral DNA, allowing it to cleave out an 11-amino-acid viral peptide, pVIc, that binds to AVP and fully activates it. Then, about 70 AVP-pVIc complexes move along the viral DNA, via one-dimensional diffusion, cleaving virion precursor proteins 3200 times to render a virus particle infectious. Late in adenovirus infection, in the cytoplasm, the cytoskeleton is destroyed. The amino acid sequence of the C terminus of actin is homologous to that of pVIc, and actin, like pVIc, can act as a cofactor for AVP in the cleavage of cytokeratin 18 and of actin itself. Thus, AVP may also play a role in cell lysis.Received 14 November 2002; received after revision 28 April 2003; accepted 30 April 2003