时由Morris水迷宫分得的青年、老年记忆正常和记忆减退鼠的脑组织分别进行突触，AChE纤维、突触素、小白蛋白神经元以及突触体钙离子浓度（[Ca2+]）、膜流动性的定量分析。结果表明老年记忆减退鼠新皮质、海马结构突触素含量、突触、胆碱能纤维、小白蛋白阳性神经元密度及突触体膜流动性较老年记忆正常和青年鼠明显降低。[Ca2+]明显增加。老年记忆正常鼠与青年鼠各项均乇显著差异。本研究提示各研究指标的异常与老年学习记忆减退密切相关。

目的和方法：X射线照射结合无血清培养诱导小鼠脾细胞凋亡，采用流式细胞术定量分析碱性成纤维细胞生长因子（bFGF）对睥细胞凋亡的抑制作用，井采用淋巴细胞转化试验[3H]-TdR掺人法检测bFGF对脾细胞的增殖效应。结果：小鼠经过不同剂量X射线照射、脾细胞再经过不同时程无血清培养，都出现了典型的亚二倍体Ap峰，1μg/mL和2μg/mL的bFCF均可降低Ap峰中的细胞数（细胞凋亡率）。并能显著增强睥细胞的增殖反应能力。结论：bFCF可抑制外周免疫器官脾细胞的凋亡、井促进其增殖。增强机体免疫力。

AIM: To obtain high-level expression of nonfusion recombinant human basic fibroblast growth factor (rhbFGF). METHODS: hbFGF cDNA was prepared from the total RNA of embryonic brain tissue. As a template, the obtained gene was used to clone nonfusion rhbFGF. New primers were employed to alter the translation initiation region (TIR) and reduce the G+C content through nucleotide change. Using pET-3C as vector, the cloned rhbFGF was expressed in BL21 (DE3). RESULTS: rhbFGF was expressed in E coli up to 30% of the total cellular protein. Cation exchange and heparin affinity chromatography were employed to purify the target protein from the supernatant of bacteria lysate. The bioactivity of the purified rhbFGF was identical with the standard bFGF. CONCLUSION: Modification of TIR is an effective means to increase nonfusion expression rate of recombinant proteins, such as rhbFGF, in E coli.

Therapeutic neovascularization for cardiovascular ischemia is a promising avenue in spite of disappointing early clinical trial results. The concept of three different mechanisms of neovascularization has served to define potential therapeutic targets such as vascular remodeling and stem cell recruitment, but it is anticipated that this will lose significance as the pleiotropic nature of angiogenic cytokines becomes fully understood. With the rapidly growing body of data on growth factors and pro-angiogenic strategies, approaches will emerge that are more effective than the ones that have been tested clinically thus far. Combinations of growth factors, for instance to stabilize vessels, or growth factors combined with cell transplants deserve more attention but will make the design of preclinical and clinical studies increasingly complex. Recent developments suggest that when using the appropriate dose and treatment regimens, even single growth factor therapy can result in stable and functional vessels. Whether gene therapy or protein therapy will be optimal for this purpose depends mainly on technical developments in vector design and production and on progress in the engineering of slow release matrix formulations for proteins. With the increasing complexity of therapeutic strategies, it remains imperative that these approaches are rationally based on fundamental and preclinical data.

In order to obtain the recombinant VPAC2 agonist efficiently by intein-mediated single column purification, a gene encoding 32-amino acids peptide was designed, synthesized and cloned into Escherichia coli expression vector pKYB. The recombinant vector pKY-ROM was transferred into E. coli ER2566 cells and the target protein was over-expressed as a fusion to the N-terminus of a self-cleavable affinity tag. After the rMROM-intein-CBD fusion protein was purified by chitin-affinity chromatography, the self-cleavage activity of the intein was induced by β-mercaptoethanol and the rMROM with the homogeneity over 95% was released from the chitin-bound intein tag. The recombinant linear rMROM competitively displaced [125I] PACAP38 on VPAC2 with a half-maximal inhibitory concentration (IC50) of 60±5 nM, whereas the IC50 of rMROM at human VPAC1 was observed up to 10 mMand no binding was detected at PAC1. rMROM stimulated the cAMP accumulation in Chinese hamster ovary (CHO) cells expressing the human VPAC2 with a half-maximal stimulatory concentration (EC50) of 0.6 nM, which was 500-fold less potent at VPAC1and had no activity on PAC1. An efficient production procedure of a novel recombinant VPAC2-selective agonist was established.

Fibroblast growth factor-2 (FGF-2) is a multifunctional polypeptide that affects many cellular functions and phenomena. The wild-type recombinant human fibroblast growth factor rhFGF-2W and the mutant C78SC96S rhFGF-2M were expressed in Escherichia coli and their products were purified. The results by the means of fluorescence spectroscopy and CD spectrums, suggested that due to its decreased hydrophobicity rhFGF-2 is not deposited as an inclusion body. The mitogenic activity of the expressed rhFGF-2M on 3T3 fibroblasts was shown to be 10-fold more than the expressed rhFGF-2W of which the biological activity was a little less than that of the standard rhbFGFW, indicating that the increased biological activity was due to the change of its secondary structure, dimerization and affinity binding to FGF receptor (FGFR).

包涵体的形成与外源基因在大肠杆菌中的表达量高度相关，在适当的范围内，降低hbFCF在表达宿主BL21（DE3）plysS中的表达，成为实现高可溶性表达的关键。在不改变氨基酸序列的条件下，对hbFCF高表达菌株突变重组子起始密码ATC下游前3个密码子的摇摆碱基进行随机回复突变，共有7种组合，合成引物PCR扩增后，克隆至表达载体pET-3e，将重组子转导BL21（DE3）plysS后，IPTC诱导表达，发现其中1株有较高可溶性和活性的菌株。可见部分降低外源蛋白的表达量可以避免与减少包涵体的形成。

AIM: To investigate the condition necessay for high-level expression of basic fibrob-last growth factor (hbFGF) in Escherichia coli, adicistron system was constructed. METHODS:1he phage T7 gene φ10 N-tenninal sequence, present in the expression plasmid pET-3c, was incolporated into the first cistron, the coding region for bFCF was incorporated into the second, and boLh were under the control of a transcnption promotor recogruzed by the bacteriophage T7 RNA polymerase. RESULTS: Recambinants were transfonned into BI21(DE3) and induced by IPTG for expression, accumulation of up to 20% of the total ceU protein of bFGF were obtained, showing bioactivity indistingushable from standard protein. CONCLUSION: It suggested that such dicistron system was effective for enhancing hbFCF expression.

采用两性离子胶体沉淀和乙醇沉淀相结合的粗提方法，经离子交换、疏水层析、亲和层析及凝胶过滤4个步骤有效地将人尿激肽原酶（hk-l）粗提物纯化，比活提高了1755倍，总得率为70%.用以慈菇蛋白酶抑制剂为配体的亲和层析纯化hk-l，效果理想，整个工艺路线适合产业化生产。纯化产物在SDS-聚丙烯酰胺凝胶电泳上为单带，高压液相色谱（HPLC）上为单峰，基质辅助激光解析电离飞行时间质谱测得分子质量为33 450u，等电聚焦测得p14.3附近，为含糖蛋白。同时测定了该酶的热稳定性和pH稳定性。纯化过程中同时分离得到另一种药用蛋白—人尿胰蛋白酶抑制剂（HUTI）。