L-硝基精氨酸对大鼠急性脑缺血损伤后炎症因子及细胞凋亡的影响

目的：观察L-硝基精氨酸（L—NA）对局灶性脑缺血损伤后炎症因子和神经细胞凋亡的影响。探讨L—NA保护脑缺血损伤组织的作用机制。方法：健康雄性SD大鼠,体重250—280g,随机分为3组（n=10）：假手术组（SH组）、缺血组（IS组）、L—NA治疗组（L—NA组）。IS、L—NA组采用线栓法制备大鼠局灶性脑缺血损伤模型。L-NA组每次腹腔注射L—NA20mg／kg,每日2次,连续3d。IS组给予等量的生理盐水。将大鼠断头取脑,采用免疫组化法检测脑组织中TNF—α表达变化,放免法检测IL-1β水平变化,流式细胞仪测定脑组织神经元凋亡率、Bcl-2蛋白、Bax蛋白表达及Bcl-2蛋白与Bax蛋白比值（Bcl-2／BaX）。结果：与SH组比较,IS组脑缺血灶范围内TNF-α表达明显增强,IL-1β水平显著升高,神经凋亡率及Bax蛋白表达升高,Bcl-2／BaX降低;与IS组比较,L—NA组脑缺血灶范围内TNF-α表达及IL-1β水平显著降低,神经凋亡率降低,Bcl-2蛋白表达及Bcl-2／BaX升高,Bax蛋白表达降低结论：L—NA通过抑制TNF-α和IL-1β的升高,增加Bcl-2蛋白表达,降低Bax蛋白表达,调节Bcl-2／Bax平衡,对脑缺血大鼠脑神经元产生一定程度的保护作用。     

A novel heterodimeric cytokine consisting of IL-17 and IL-17F regulates inflammatory responses

CD4＋ helper T （TH） cells play crucial roles in immune responses. Recently a novel subset of TH cells, termed THIL-17, TH 17 or inflammatory TH （THi）, has been identified as critical mediators of tissue inflammation. These cells produce IL-17 （also called IL-17A） and IL-17F, two most homologous cytokines sharing similar regulations. Here we report that when overexpressed in 293T cells, IL-17 and IL-17F form not only homodimers but also heterodimers, which we name as IL-17A/F. Fully differentiated mouse THi cells also naturally secrete IL-17A/F as well as IL-17 and IL-17F homodimeric cytokines. Recombinant IL-17A/F protein exhibits intermediate levels of potency in inducing IL-6 and KC （CXCL 1） as compared to homodimeric cytokines. IL-17A/F regulation of IL-6 and KC expression is dependent on IL-17RA and TRAF6. Thus, IL-17A/F cytokine represents another mechanism whereby T cells regulate inflammatory responses and may serve as a novel target for treating various immune-mediated diseases.     

Deficiency of DNA fragmentation factor 45 results in reduced oocyte apoptosis in response to doxorubicin

Dear Editor： Apoptosis plays a prominent role in ovarian development and function. During follicle development, the vast majority of follicles undergo atresia as a consequence of apoptosis of constituent oocyte or follicular cells, or both, failing to complete the maturation process. Atresia occurs at all stages of follicular development during the growth and development of follicles.[第一段]     

Targeted gene disruption by use of a group 11 intron （targetron） vector in Clostridium acetobutylicum

Dear Editor： Clostridium acetobutylicum, a gram-positive, anaerobic, spore-forming bacterium, is capable of using a wide variety of carbon sources to produce acetone, butanol and ethanol. To improve solvent productivity of C. acetobutylicum, metabolic engineering is considered as a useful tool in developing strains with industrially desirable character-istics. However, to date, there are few useful methods for genetic manipulation of C. acetobutylicum, especially for gene disruption. To our knowledge, two types of vectors, including non-replicative and replicative integrative plasmids, have been developed for gene-inactivation in C. acetobutylicum. By using non-replicative integrative plasmids, buk and solR genes of C. acetobutylicum were inactivated [1,2]. However, due to their low frequencies of transformation and recombination, the non-replicative integrative plasmids are usually transformed at less than 1 integrative transformant per mg plasmid DNA. To obtain the integrative mutant, it may require higher transformation frequencies up to 10^5, but the typical transformation fre-quencies were reported at 10^3 [3].[第一段]     

Cancer research in China： a Special Focus by Cell Research

The year of 2007 is the 100th anniversary of the American Association for Cancer Research （AACR）, ＂marking a century of progress in saving lives through cancer research＂ （quote from the AACR website）. Despite the great progress, cancer remains a top threat to human health. Continued research will be essential to winning the battle against cancer, as only through continued research would it be possible to gain better understandings, generate deeper insights, uncover new preventive and therapeutic strategies, design and test more effective treatments, and ultimately, save more lives.     

Generation and selection of immunized Fab phage display library against human B cell lymphoma

The approval of using monoclonal antibodies as a targeted therapy in the management of patients with B cell lymphoma has led to new treatment options for this group of patients. Production ofmonoclonal antibodies by the traditional hybridoma technology is costly, and the resulting murine antibodies often have the disadvantage of triggering human anti-mouse antibody （HAMA） response. Therefore recombinant Fab antibodies generated by the phage display technology can be a suitable alternative in managing B cell lymphoma. In this study, we extracted total RNA from spleen cells of BALB/c mice immunized with human B lymphoma cells, and used RT-PCR to amplify cDNAs coding for the κ light chains and Fd fragments of heavy chains. After appropriate restriction digests, these cDNA fragments were successively inserted into the phagemid vector pComb3H-SS to construct an immunized Fab phage display library. The diversity of the constructed library was approximately 1.94× 10^7. Following five rounds of biopanning, soluble Fab antibodies were produced from positive clones identified by ELISA. From eight positive clones, FabC06, FabC21, FabC43 and FabC59 were selected for sequence analysis. At the level of amino acid sequences, the variable heavy domains （VH） and variable light domains （VL） were found to share 88-92% and 89-94% homology with sequences coded by the corresponding murine germline genes respectively. Furthermore, reactivity with membrane proteins of the B cell lymphoma was demonstrated by immunohistochemistry and western blotting. These immunized Fab antibodies may provide a valuable tool for further study of B cell lymphoma and could also contribute to the improvement of disease therapy.     

Centromere cohesion： regulating the guardian

Mitosis, a process in which duplicated chromosomes are segregated into two daughter cells, is the most spectacular event in cell cycle. In mitosis, cells undergo drastic rearrangement of cytoskeleton, lipid and chromosomes under amazingly strict temporal and spatial control so that genetic information is faithfully transmitted to daughter cells.[第一段]     

Niche regulation of corneal epithelial stem cells at the limbus

Among all adult somatic stem cells,those of the corneal epithelium are unique in their exclusive location in a definedlimbai structure termed Palisades of Vogt.As a result,surgical engraftment oflimbal epithelial stem cells with or withoutex vivo expansion has long been practiced to restore sights in patients inflicted with limbal stem cell deficiency.Neverthe-less,compared to other stem cell examples,relatively little is known about the limbal niche,which is believed to play apivotal role in regulating self-renewal and fate decision of limbal epithelial stem cells.This review summarizes relevantliterature and formulates several key questions to guide future research into better understanding of the pathogenesis oflimbal stem cell deficiency and further improvement of the tissue engineering of the corneal epithelium by focusing onthe limbal niche.     

bFGF对卵巢癌CAOV3细胞Bcl-2、Bcl-xl、Bax、Bad表达的影响

目的 研究bFGF调控卵巢癌CAOV3凋亡的信号通路及对Bcl-2、Bcl-xl、Bax、Bad表达的影响.方法 无血清饥饿诱导细胞凋亡.分为饥饿对照、bFGF、bFGF ＋ PD98059、bFGF ＋ Wortmannin组.流式细胞术、DNA Ladder检测细胞凋亡;Western印迹法检测ERK、PKB、Bad活性以及Bcl-2、Bax表达,RTPCR检测Bcl-2、Bcl-xl mRNA变化.结果 bFGF促进p-ERK、p-PKB、p-Bad、Bcl-2表达,抑制Bax表达及饥饿诱导的细胞凋亡.激酶抑制剂PD98059可抑制bFGF对ERK、Bcl-2、Bax的调节作用,Wortmannin可抑制bFGF对PKB、Bad、Bax的调控作用,二者均可阻断bFGF对凋亡的抑制作用.bFGF对Bcl-xl表达无影响.结论 bFGF可能通过激活MEK/ERK、P13K/PKB信号途径通路调节Bcl-2、Bax、Bad表达,抑制饥饿诱导的卵巢癌CAOV3细胞凋亡.     

南方红豆杉和东北红豆杉中的单体化合物对乳腺癌细胞增殖的影响

本研究采用MTT法,研究南方红豆杉和东北红豆杉中的10种不同结构类型的单体化合物对乳腺癌MCF-7细胞增殖的影响。结果表明,化合物1~10(10-9~10-5mol/L)处理MCF-7细胞48和72 h后,仅化合物4在10-7、10-6和10-5mol/L浓度对MCF-7细胞增殖均有明显抑制作用,抑制率分别为29.8%、46.4%、51.8%和43.6%、61.2%、63.2%,与紫杉醇抑制细胞增殖的活性相近,且在24~72 h范围内具有时间依赖性;化合物2仅在10-5mol/L具有明显抑制细胞增殖的作用,抑制率为44.4%和49.6%。因此,10种不同结构类型的单体化合物中,仅baccatin III类化合物2、4对MCF-7细胞增殖具有较强的抑制作用,其中化合物4作用最强,活性与紫杉醇相近。