Long-term fate of human telencephalic progenitor cells grafted into the adult mouse brain: effects of previous amplification in vitro

We assessed the developmental potential of human telencephalic progenitor cells, with and without previous amplification in vitro, following grafting into the nonlesioned adult mouse CNS. Cell suspensions, shown to contain neuroepithelium-like and neuroblast-like cells, were injected into the subventricular zone (SVZ) and the striatum. These two regions were selected for comparative studies because one, the SVZ, is mitotically active, whereas the other, the striatum, is mitotically inactive. In situ hybridization with a human-specific Alu probe showed that the cells survived for up to 30 weeks in both targets and migrated away from the injection site. Fresh cells continued to proliferate and gave rise to very extended grafts before differentiating into neurons and glia. We further show that, when grown in vitro prior to grafting, human cells acquired new properties: Their proliferation was very limited, and they differentiated more rapidly. This study therefore provides new information about the use of these cells, which are a potential tool for both cellular and gene therapy.     

Oestrogens,via transforming growth factor alpha,modulate basic fibroblast growth factor synthesis in hypothalamic astrocytes: in vitro observations

The data presented here show that, in cultures of type 1 astrocytes obtained from the hypothalamus of neonatal female rat, 17beta-oestradiol is able to increase both the mRNA and the protein levels of basic fibroblast growth factor (bFGF). In particular, after 24 h of exposure to 17beta-oestradiol (10(-9) and 10(-10) m), an increase of messenger levels of bFGF appears in hypothalamic type 1 astrocytes. Similarly, an induction of bFGF protein is also evident at this time of exposure. The effect on the mRNA and protein levels of bFGF is blocked by the presence in the medium of an antibody raised against the transforming growth factor alpha (TGFalpha) receptor. This observation indicates that, TGFalpha, whose synthesis is modulated by oestrogens in hypothalamic astrocytes and which is able to increase, both the mRNA and the protein levels of bFGF in our experimental model, may act as the mediator of the oestrogenic induction of bFGF. Hypothalamic astrocytes, together with hypothalamic neurones synthesizing and secreting luteinizing hormone-releasing hormone (LHRH), form the LHRH network in conjunction with other neuronal systems. Gonadal steroids in general, and oestrogens in particular, play an important role in the control of the activity of this network. In addition, bFGF and TGFalpha, two growth factors released from astrocytes, are able to influence the activity of LHRH neurones. The present observations suggest that oestrogens may also act on LHRH neurones in an indirect fashion (i.e. by modulating the expression of bFGF and TGFalpha in glial cells).     

Postischemic administration of basic fibroblast growth factor improves sensorimotor function and reduces infarct size following permanent focal cerebral ischemia in the rat

Basic fibroblast growth factor (bFGF) is a polypeptide with potent trophic and protective effects on the brain. bFGF has been reported to exert neuroprotection against a wide variety of insults, including ischemic neuronal injury. To date, animal models of focal ischemia have not been translated to efficacy in stroke clinically with respect to testing of neuroprotective agents. Because functional outcome is the measurement of efficacy for putative neuroprotective agents in the clinic, we sought to evaluate the functional consequences of bFGF administration in rats subjected to focal ischemia. In this study, we assessed the effects of bFGF on functional outcome as well as infarct size in rats subjected to severe cerebral ischemia by permanent occlusion of the middle cerebral artery (MCAO). Male Sprague-Dawley rats were subjected to permanent MCAO by the intraluminal filament technique. Two hours following occlusion, rats were infused intravenously with either bFGF, at a dose of 150 microg/kg, or vehicle alone. Functional sensorimotor impairment, which was assessed by the accelerating rotarod test, was recorded at baseline and compared to performance assessed at 24 h after MCAO. Permanent occlusion of the MCA caused marked impairment in rotarod performance in both groups. Treatment of rats with bFGF showed a significant 46% improvement in rotarod fall latency when compared with that from the animals treated with vehicle alone. The volume of cortical infarction was significantly reduced by 32% as a function of bFGF treatment. These results suggest that the delayed intravenous administration of bFGF improves sensorimotor function as well as reduces infarct size following permanent focal ischemia in rat.     

Immunohistochemical profile of cystosarcoma phyllodes of the breast: a study of 23 cases

Cystosarcoma phyllodes (CP) of the breast is a rare biphasic tumor composed of benign epithelium and a spindle cell stroma. Biologic behavior of CP cannot be predicted with certainty on the basis of morphologic criteria only. We studied immunohistochemical expression of basic fibroblastic growth factor (bFGF), urokinase, Ki67, p53 protein, and microvessel density in stromal and epithelial components of 14 low-grade CP (LCP) and 9 high-grade CP (HCP). bFGF was more often positive in LCP than in HCP. The stroma was positive for bFGF in 86% of LCP and 67% of HCP, and the epithelium was positive in 64% of LCP and 14% of HCP. Urokinase was positive in stromal cells of 86% of LCP and 93% of HCP. The epithelial positivity for urokinase in both groups resembled closely that of the stroma. p53 protein was more often positive in stromal cells of HCP (67%) than in LCP (50%). Ki67 was positive in the stroma of 43% of LCP and 89% of HCP and in the epithelium of 14% of LCP and 33% of HCP. There was no significant difference in microvessel density (MVD) in low- and high-grade lesions. Our study demonstrates that stromal Ki67 and p53 immunohistochemical positivity are more often associated with high-grade tumors. The positive immunostaining for bFGF, urokinase, Ki67, and p53 in stroma and epithelium of the majority of CP supports the existence of epithelial-stromal interactions and recognizes epithelium as an integral part of this tumor.     

EGF,bFGF单独及联合应用对原代培养的兔角膜缘干细胞增殖的影响

目的探讨EGF,bFGF及二者共同作用对培养的兔角膜缘干细胞增殖的影响。方法用DMEM和F12(1:1)培养基,20%胎牛血清,加入不同浓度EGF,bFGF和二者的混合液,采用MTT法检测细胞的OD值。结果EGF的5~160ng/mL各浓度组增殖效应均明显大于对照组(P<0.05),10ng/mL组明显大于其它各浓度组(P<0.01),bFGF各浓度组从5~80ng/mL各组均明显大于对照组(P<0.025),20ng/mL各组明显大于与其它各组(P<0.025);EGF与bFGF混合液各浓度组从5~80ng/mL各组增殖效应均明显大于对照组及单独使用EGF或bFGF组(P<0.05),EGF10ng/mL+bFGF20ng/mL组促增殖效果最强(P<0.025)。结论EGF,bFGF对培养的兔角膜缘干细胞有促增殖作用,此作用均呈剂量依赖性,EGF和bFGF共同表现为明显的促增殖作用。     

贝复舒在治疗真菌性角膜溃疡中的应用

目的观祭治疗具酉性角膜溃疡中应用贝复舒的效果。方法真菌性角膜溃疡患者100例100眼随机分戍治疗组70眼和对照组30眼，治疗组应用贝复舒，对照组应用角膜宁，疗程10d。对比观察2组角膜创面愈合与角膜缘血管翳形成情况。结果治疗组局部创面1周内全部愈合，术后6月3眼形成角膜缘血管翳；对照组1周内22眼愈合，术后6月20眼形成角膜缘血管翳。2者比较差异显著（P〈0．01）。结论贝复舒在治疗真菌性角膜溃疡中具有显著效果。[眼科新进展2005；25（5）：454—455]     

Basic fibroblast growth factor expression in isolated small human ovarian follicles

Basic fibroblast growth factor (bFGF) is involved in cell proliferation, differentiation, and angiogenesis. It has long been known that bFGF acts as a powerful mitogen for various mammalian granulosa cells in culture. To investigate the possible involvement of bFGF expression in follicle initiation and growth in vivo, we performed nested RT-PCR on ovarian cortical biopsies and quantitative PCR on human follicle populations isolated by laser capture microdissection. Using morphological criteria, follicles were characterized as putative non-growing, primary, or small secondary. RNA was extracted from samples, reverse-transcribed, and relative gene expression levels determined with TaqMan real-time PCR, using 18S rRNA as the endogenous control. Results confirmed bFGF expression in human adult ovarian cortex, and in the isolated follicles a down-regulation of bFGF mRNA was evident as small follicles develop. This study demonstrates a possible relationship between bFGF mRNA expression and follicle development.