银杏提取液对随意皮瓣bFGF表达的影响

目的研究银杏提取液能否通过刺激随意皮瓣碱性成纤维细胞生长因子(bFGF)的表达来促进皮瓣的成活。方法取健康SD大鼠24只分为2组,构建大鼠背部缺血随意皮瓣,实验组大鼠每天ip银杏提取液100 mg/kg,对照组大鼠每天ip等量生理盐水。术后7 d通过大体观察皮瓣成活,血管造影术观察皮瓣血管显影程度,HE染色观察皮瓣病理改变情况,免疫组化法检测bFGF的表达情况。结果实验组皮瓣的存活率为(67.07±2.66)%,明显高于对照组皮瓣的存活率(48.64±2.09)%(P<0.01)。实验组皮瓣远端的血管显影程度、bFGF的表达均高于对照组(P<0.01)。结论银杏提取液能通过刺激bFGF的表达来促进局部血管形成,从而在大体上促进高皮瓣的成活。     

GDF‐5/7 and bFGF activate integrin α2‐mediated cellular migration in rabbit ligament fibroblasts

Cellular activities responding to growth factors are important in ligament healing. The anterior cruciate ligament (ACL) has poor healing potential compared to the medial collateral ligament (MCL). To assess the differences, we investigated the proliferation, migration, adhesion, and matrix synthesis responding to growth factors in rabbit ACL and MCL fibroblasts. ACL cell proliferation to basic fibroblast growth factor (bFGF), bone morphogenetic protein‐2, growth and differentiation factor (GDF)‐5, and GDF‐7 treatment was similar to that of MCL cells. GDF‐5 enhanced Col1a1 expression in ACL and MCL fibroblasts up to 4.7‐ and 17‐fold levels of control, respectively. MCL fibroblasts showed stronger migration activities in response to bFGF and GDF‐5 than ACL cells. GDF‐5/7 and bFGF also changed the stress fiber formation and cellular adhesion by modulating the distribution of integrin α2. Functional blocking analyses using anti‐integrin α2 antibodies revealed that cellular migration responding to growth factors depended on the integrin α2‐mediated adhesion on type I collagen. The expression of integrin α2 was also increased by growth factors in both cells. Our results demonstrate that GDF‐5/7 and bFGF stimulate cellular migration by modulating integrin α2 expression and integrin α2‐dependent adhesion, especially in MCL fibroblasts. These findings suggest that the different healing potential between ACL and MCL may be caused by different cellular behavior in the integrin α2‐mediated cellular migration in response to growth factors. © 2009 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 28:225–231, 2010     

Expression of bFGF/FGFR-1 and vascular proliferation related to clinicopathologic features and tumor progress in localized prostate cancer

Microvessel density (MVD) has been associated with progression of prostate cancer. Although basic fibroblast growth factor (bFGF) is a known endothelial mitogen, the prognostic role of bFGF and its receptor FGFR-1 in prostate cancer has been controversial. The aim of our study was to examine the tissue distribution and prognostic significance of bFGF, FGFR-1, and microvascular proliferation. Sections from 104 radical prostatectomy specimens were examined by factor VIII/Ki-67 staining for proliferating capillary index (PCI) and MVD, and tissue microarray sections were immunostained for bFGF and FGFR-1. Increased PCI (median 0.49%) was related to strong stromal expression of bFGF (P=0.003) but was without prognostic impact. Strong bFGF staining was associated with well-differentiated tumors, no capsular penetration, low serum-prostate-specific antigen (s-PSA), low tumor cell proliferation, and increased time to biochemical failure (P=0.007), and was of independent prognostic importance in multivariate survival analysis. bFGF expression in vessels was associated with low MVD (P=0.0003). In contrast, strong tumor cell FGFR-1 expression was related to high preoperative s-PSA. Thus, increased stromal and vessel bFGF was associated with less aggressive tumors. Our findings indicate a complex relationship between bFGF/FGFR-1 expression and prognosis of prostate cancer. Vascular proliferation revealed no prognostic impact in this study.     

bFGF与VEGF抗原表位筛选及复合肽的表达鉴定

目的筛选bFGF与VEGF抗原表位,构建复合肽进行原核表达,并对复合肽免疫原性及抑瘤效果进行研究。方法生物信息学方法分别预测bFGF和VEGF抗原表位,筛选优势表位,通过柔性Linker连接各表位构建复合肽,合成引物,重叠延伸PCR(gene splicing by overlap extension PCR,SOE PCR)方法合成复合肽cDNA序列,克隆入原核表达载体pET32a进行表达,表达产物进行SDS-PAGE和Western-Blot鉴定,Ni-NTA柱纯化复合肽并免疫小鼠,间接ELISA法测定抗bFGF抗体和抗VEGF抗体滴度。构建黑色素移植瘤C57BL/6小鼠模型,研究复合肽免疫后肿瘤生长情况。结果生物信息学方法筛选获得3个bFGF表位和2个VEGF表位连同一个噬菌体多肽库筛选表位构建成复合肽,SOEPCR方法成功合成复合肽cDNA序列,构建原核表达载体表达bFGF与VEGF复合肽,纯化的复合肽免疫小鼠产生抗bFGF抗体和抗VEGF抗体;并且体内能有效抑制黑色素瘤生长。结论生物信息学方法筛选bFGF和VEGF获得的抗原表位构建的复合肽在体内同时激发抗bFGF抗体和抗VEGF抗体,并且能抑制肿瘤...     

Time- and dose-dependent mitogenic effect of basic fibroblast growth factor combined with different bone graft materials: an in vitro study

OBJECTIVES: In periodontal regeneration, the growth factor concentrations and the delivery system used are of great importance. In an attempt to assess the mitogenic effect of basic fibroblast growth factor (bFGF) on periodontal ligament (PDL) cells combined with different bone replacement materials, two allografts of cortical (DFDBA) and cancellous (DFBA) bone and an anorganic bovine material with a synthetic peptide (ABM P-15) were used. The purpose of this study was to evaluate the in vitro mitogenic effect of different doses of bFGF alone or in combination with DFDBA, DFBA and ABM P-15 on human PDL cells in a time-dependent mode. MATERIAL AND METHODS: PDL cell cultures were derived from the mid-root of four maxillary premolars. Cells were grown and reached confluence. On day 2 of quiescence, new medium was added along with (1) 1, 5, 10 and 25 ng/ml of bFGF alone, (2) 10 mg of DFDBA, DFBA and ABM P-15 alone and (3) their combination. The mitogenic effect was determined at 24 and 48 h of culture by using a hemocytometer chamber. The cells were counted under a phase contrast microscope. RESULTS: The results revealed that bFGF at the highest concentrations and after 48 h exerted a significant mitogenic effect on PDL cells, and also DFDBA and DFBA supported cell proliferation. Furthermore, DFDBA and DFBA enriched with bFGF had a significant mitogenic effect after 48 h of culture. ABM P-15 with 10 and 25 ng/ml of bFGF up-regulated PDL cell proliferation after 48 h of incubation. CONCLUSIONS: The findings of this study demonstrate the beneficial role of bFGF combined with DFDBA and DFBA as carriers in periodontal repair.     

Multifunctional implantable particles for skin tissue regeneration: preparation, characterization, in vitro and in vivo studies

The transplantation of cell-polymer constructs has been developed as a novel approach to curing tissue defects. However, a number of methodological problems remain to be solved, including the loss of a proper cellular milieu, the relatively long period of culture time and the complexity of the application. The aim of the present article is to evaluate the feasibility of porous gelatin-based implantable particles as a novel strategy for delivery of cultured cells and bioactive molecules to correct dermal defects. For this purpose, implantable porous gelatin particles (100-230 microm) encapsulating proliferative growth factors were prepared and characterized, and their influence on fibroblasts was assessed. In vivo examinations were undertaken to observe guided dermal tissue regeneration after the transplantation of the implantable particles. Our results indicate the feasibility of transplanting multifunctional implantable particles as a culture substrate, as a protein transplantation vehicle or as a biodegradable implant for skin regeneration, thus giving an indication of the possible applications in tissue engineering.     

rhFGF与rhEGF分期使用修复深Ⅱ度烧伤创面的临床观察

目的:观察分期使用重组牛碱性成纤维细胞生长因子(rhFGF)与重组人表皮细胞生长因子(rhEGF)治疗深Ⅱ度烧伤创面愈合的临床疗效。方法:随机选择同一患者4处深度相同、部位相应的创面进行自身对照,4处创面分别标记为A、B、C、D,20例深Ⅱ度烧伤患者共80个创面,相应归为A、B、C、D组:A组创面采用rhEGF治疗,B组创面采用rhFGF治疗,C组创面先使用rhFGF,10d后使用rhEGF治疗,D组创面为0.9%氯化钠对照组。观察创面完全愈合所需要的时间、创面愈合质量,以及全身和局部反应。结果:A组的平均愈合时间为(18.20±1.67)d,B组为(19.15±2.01)d,C组为(16.45±1.57)d,D组为(23.65±2.41)d。A、B、C组的平均愈合时间较D组短,C组的平均愈合时间较A、B组短,差异均具有统计学意义(P<0.05);A组与B组相比,愈合时间无显著性差异(P>0.05)。A、B、C组的平均愈合质量比D组好。各组治疗期间未见明显的全身或局部不良反应。结论:分期使用rhFGF和rhEGF能有效促进深Ⅱ度烧伤创面的愈合和修复,缩短愈合时间。     

脑络通方对光化学诱导脑缺血大鼠脑内bFGF影响的实验研究

目的研究脑络通方对光化学诱导脑缺血大鼠碱性成纤维细胞生长因子(bFGF)表达的影响.方法用免疫组化方法,观察光化学诱导的脑缺血大鼠第5天时,各组脑内缺血灶、缺血灶周围、大脑皮层、海马、小脑等部位bFGF的表达.结果脑缺血5天后,脑内bFGF主要在缺血灶周围、海马部位表达,阳性细胞主要以神经胶质细胞、神经元细胞为主,治疗组表达比模型组明显增加.结论脑络通方可通过提高缺血灶周围和海马bFGF的表达,从而促进缺血后神经元的存活,参与脑内的损伤修复和神经再生.     

电针对脾虚型幼鼠侧脑室下区碱性成纤维细胞生长因子(bFGF)表达的影响

目的:探讨电针对脾虚证大鼠侧脑室下区碱性成纤维细胞生长因子表达的影响。方法:健康SD雄性大鼠96只,体重(50±5)g,鼠龄为4周龄,随机选取32只作为正常组,其余64只造模。造模成功后随机分为模型组、电针组,每组各32只。3组再分别随机分为7天组、14天组、28天组和49天组4个亚组,每亚组8只。用利血平腹腔注射联合大黄灌胃造SD雄性幼鼠脾虚证模型,造模期14天。造模结束后,电针组大鼠针刺双侧足三里和三阴交穴,以疏密波刺激20min,每天1次,治疗7、14、28、49天后处死,采用免疫组织化学法观察侧脑室下区碱性成纤维细胞生长因子数量表达的变化。结果:电针治疗7天,针刺组碱性成纤维细胞生长因子数量明显较模型组和正常组升高(P0.01);电针治疗14天,其数量有所回落,与同期模型组相当,但仍比正常组高(P0.05),这一变化可持续至28天;电针治疗49天,针刺组碱性成纤维细胞生长因子数量仍较同期模型组和正常组高(P0.01)。结论:早期介入电针治疗可以更早地能对脾虚证所造成对脑功能损害起到保护作用。同时,持续的电针治疗可以在相当长的一段时期内保持碱性成纤维细胞生长因子数量有较高的表达。