Effect of electro-acupuncture on basic fibroblast growth factor protein and mRNA expression in hippocampal dentate gyrus of spleen deficiency rats

BACKGROUND: Spleen deficiency in traditional Chinese medicine refers to the functional disorder of spleen,pancreas,intestines,and nervous system in modern medicine.OBJECTIVE: To test whether electro-acupuncture could alter basic fibroblast growth factor (bFGF) protein and mRNA expression in the hippocampal dentate gyrus of spleen deficiency rats.DESIGN,TIME AND SETTING: The randomized,controlled,in vivo animal experiment was performed at the National Level-B Laboratory of Clinical Cell Molecule and Biology in Shenzhen Hospital of Traditional Chinese Medicine,between March and November in 2008.MATERIALS: Reserpine injection was produced by Guangdong Bangmin Pharmaceutical Co.Rhubarb extract granule preparation was produced by Guangdong Yifang Pharmaceutical.Huanqiu Brand sterile acupuncture pin was provided by Suzhou Acupuncture Supplies,China.Huatuo Brand electroacupuncture instrument (type SDZ-II) was purchased from Suzhou Medical Appliance Factory,China.METHODS: A total of 96 male Sprague Dawley rats were randomly assigned to control (n = 32) and induction (n = 64) groups.Spleen deficiency was induced via intraperitoneal injection of reserpine and intragastric administration of rhubarb.The successful models were randomized into two groups: model and electro-acupuncture,with 32 rats in each group.Elec-tro-acupuncture was administered at Zusanli (ST 36) and Sanyinjiao (SP 6) acupoints using a condensation wave and rarefaction (condensation wave 15 Hz) at a strength of 6–15 V for 20 minutes,once per day.The appearance of a slight shiver in the corresponding locus was taken as the standard.According to electro-acupuncture time points,each group was assigned to four subgroups at 7,14,28,and 49 days,respectively,with eight rats in each subgroup.Immunohistochemical staining,image analysis,and reverse-transcription polymerase chain reaction were performed at different time points.MAIN OUTCOME MEASURES: bFGF protein and mRNA expression in the hippocampal dentate gyrus of spleen deficiency rats.RESULTS: After 7 days of electro-acupuncture therapy,bFGF protein and mRNA expression significantly increased compared with the model and control groups (P < 0.05).After 14 days,bFGF protein and mRNA expression decreased until 28 days,where levels were then equal to the model group and greater than the control group (P < 0.05).After 49 days,the above indices remained increased in the electro-acupuncture group compared to the model and control groups (P < 0.05).CONCLUSION: Continuous electro-acupuncture maintained a high level of bFGF protein and mRNA expression in the hippocampal dentate gyrus of spleen deficiency rats.

Effect of charge at an amino acid of basic fibroblast growth factor on its mitogenic activity

The amino acid at the 119th position of human basic fibroblast growth factor(hbFGF),lysine(K119),is a critical component for its mitogenic activity.However,little is known about the effects of the characteristics of this residue including charge on the mitogenic activity of hbFGF.Herein,this basic residue was replaced with neutral glutamine residue and acidic glutamic acid residue to construct mutants hbFGF^(K119Q) and hbFGF^(K119E),respectively.The mutants were produced by BL21(DE3)/pET3c expression sys...

mRNA Expression of Basic Fibroblast Growth Factor from A Single Intratracheal Instillation of Papain induced Emphysema in Rats

The relations between mRNA expression of basic fibroblast growth factor (bFGF) and the changes in collagen Ⅰand collagen Ⅲ in pulmonary tissues from a single intratracheal instillation of papain induced emphysema in rats were investigated. Wistar rats ( n =42) were randomly divided into normal group and emphysema model 1, 3, 5, 7, 15, 30 day groups ( n =6 in each group). The rat model of emphysema was induced by a single intratracheal instillation of papain. The results of immunohistochemistry SABC and in situ hybridization with bFGF probe were quantitatively analyzed to examine the changes of collagen Ⅰand collagen Ⅲ and bFGF mRNA expression in lung tissues and the percent of positive expression of bFGFmRNA in alveolar macrophages. The results were as follows: (1) In the emphysema model groups the optical densities of collagen Ⅰand collagen Ⅲ began to increase after 3 days, reached the highest at the 7 th day, and began to reduce at the 15 th day; (2) No expression of bFGFmRNA in pulmonary tissues was detectable in the normal group. The positive expression of bFGFmRNA was detectable in lung tissues one day after the intratracheal instillation of papain. The average optical densities reached the peak (41.895±7.017) at the 7th day, significantly higher than in the normal group (0.581±0.139, P <0.01). The positive expression of bFGFmRNA in lung tissues began to reduce at the 15th day; (3) Positive expression of bFGFmRNA in alveolar macrophages of instillation papain rats was detectable 3 days after the intratracheal instillation of papain, and reached the highest at the 7th day with the percent of positive expression of bFGF mRNA in alveolar macrophages being 70.13±11.21, higher than in the normal group (5.12±0.18, P <0.01); (4) The expression of bFGF mRNA in the lung tissues and macrophages was postively related with the changes in collagen Ⅰ and collagen Ⅲ ( P <0.01 or P <0.05) respectively. It was suggested that the up regulation of bFGF mRNA expression during the development of emphysema can lead pulmonary interstitial fibrosis, which may take part in the injury and repair and the lung tissue reconstruction.

Successful Management of Wound Dehiscence after Total Knee Arthroplasty by Topically Using Recombinant Basic Fibroblast Growth Factor

Wound complications are estimated to be affected in about 9% of TKA patients, which may increase the risk of deep periprosthetic infection and results in re-operation, joint fusion, or amputation. Here we have reported a female patient who suffered wound rupture due to early post-operation mobilization and weight-bearing. The wound dehiscence was successfully managed by applying recombinant basic fibroblast growth factor-2 and anti-infective treatment without removing prosthetic joint.

Effect of the implant composite of poly lactide-co-glycolide and bone mesenchymal stem cells modified by basic fibroblast growth factor on injured spinal cord in rats

Objective To investigate the effect of the implant composite of poly lactide-co-glycolide(PLGA)and bone mesenchymal stem cells (BMSCs) modified by basic fibroblast growth factor (bFGF) on injured spinal cord in rats.Methods Two hundred and

Effect of intestinal ischemia-reperfusion on expressions of endogenous basic fibroblast growth factor and transforming growth factor β in lung and its relation with lung repair

AIM To study the changes of endogenoustransforming growth factor β(TGFβ)and basicfibroblast growth factor(bFGF)in lung followingintestinal ischemia and reperfusion injury andtheir effects on lung injury and repair.METHODS Sixty Wistar rats were divided intofive groups,which underwent sham-operation,ischemia(45 minutes),and reperfusion(6,24and 48 hours,respectively)after ischemia(45minutes).Immunohistochemical method wasused to observe the localization and amounts ofboth growth factors.RESULTS Positive signals of both growthfactors could be found in normal lung,mainly inalveolar cells and endothelial cells of vein.Afterischemia and reperfusion insult,expressions ofboth growth factors were increased and theiramounts at 6 hours were larger than those ofnormal control or of 24 and 48 hours after insult.CONCLUSION The endogenous bFGF and TGF βexpression appears to be up-regulated in thelung following intestinal ischemia andreperfusion,suggesting that both growth factorsmay be involved in the process of lung injury andrepair.

Effect of basic fibroblast growth factor(bFGF) on the treatment of exposure of the orbital implants

Objective: To evaluate the efficacy and the indication of basic fibroblast growth factor (bFGF) in the treatment of exposure of orbital implants. Design: Retrospective and observational case series. Methods: We reviewed 41 patients (41 eyes) suffering exposure of orbital implants from Jan. 2000 to June 2006. The study group patients with mild exposure received com-bined treatment with bFGF and antibiotic drops, and while the control group patients with mild exposure were treated with anti-biotic drops only. The study group patients with moderate and severe exposure received combined treatment with bFGF and antibiotic drops, and after 2 months they were subjected to amniotic membrane transplantation, while the control group patients with moderate and severe exposure underwent amniotic membrane transplantation after using antibiotic drops. Observation of the growth of conjunctival epithelium and comparison of the healing rate of the two groups. Results: The healing rates of the mild, moderate and severe exposure study group were 100% and 92.3%. The healing rates of the mild, moderate and severe exposure control group were 55.6% and 66.7% respectively. The difference of the healing rates of the mild exposure study group and the control group was significant (P=0.033). And the difference of the healing rates of the moderate and severe exposure study group and the control group was not significant (P=0.167). Conclusion: bFGF may promote obviously the healing of orbital implant exposure, particularly it can be the first choice for the treatment of mild degree exposure. For the moderate and severe cases, it can be administered before surgical repair to enhance neovascularization and will tend to increase the success rate of surgical repair.

Basic fibroblast growth factor attenuates the degeneration of injured spinal cord motor endplates

The distal end of the spinal cord and neuromuscular junction may develop secondary degeneration and damage following spinal cord injury because of the loss of neural connections. In this study, a rat model of spinal cord injury, established using a modified Allen's method, was injected with basic fibroblast growth factor solution via subarachnoid catheter. After injection, rats with spinal cord injury displayed higher scores on the Basso, Beattie and Bresnahan locomotor scale. Motor function was also well recovered and hematoxylin-eosin staining showed that spinal glial scar hyperplasia was not apparent. Additionally, anterior tibial muscle fibers slowly, but progressively, atrophied. Immunohistochemical staining showed that the absorbance values of calcitonin gene related peptide and acetylcholinesterase in anterior tibial muscle and spinal cord were similar, and injection of basic fibroblast growth factor increased this absorbance. Results showed that after spinal cord injury, the distal motor neurons and motor endplate degenerated. Changes in calcitonin gene related peptide and acetylcholinesterase in the spinal cord anterior horn motor neurons and motor endplate then occurred that were consistent with this regeneration. Our findings indicate that basic fibroblast growth factor can protect the endplate through attenuating the decreased expression of calcitonin gene related peptide and acetylcholinesterase in anterior horn motor neurons of the injured spinal cord.

The effect of basic fibroblast growth factor on regeneration in a surgical wound model of rat submandibular glands

This study developed an animal model of surgically wounded submandibular glands(SMGs)and investigated the effects of collagen gel with basic fibroblast growth factor(b FGF)on tissue regeneration of surgically wounded SMGs in vivo.The animal model was produced by creating a surgical wound using a 3-mm diameter biopsy punch in SMGs.The wound was filled with collagen gel with b FGF(b FGF group)or without b FGF(control group).In the animal model of surgically wounded SMGs,salivary glands without scar tissue around the wound area were observed with smaller areas of collagen gel.Small round and spindle-shape cells invaded the collagen gel in both groups after operation day(AOD)5,and this invasion dramatically increased at AOD 7.Host tissue completely replaced the collagen gel at AOD 21.The invading immune cells in the group treated with collagen gel with b FGF were positive for vimentin,a-smooth muscle actin(a SMA),CD49f,c-kit and AQP5 at AOD 7.Similarly,the m RNA expression of vimentin,a SMA,CD49f,keratin19 and AQP5 was also increased.This study suggests that the use of collagen gels with b FGF improves salivary gland regeneration.

Experimental research of curing sciatic nerve injury using nerve growth factor and basic fibroblast growth factor combination

AIM:To explore the co effect of nerve growth factor (NGF) and basic fibroblast growth factor (bFGF) on the sciatic nerve after injury. METHODS: 96 rats were divided into normal saline group, NGF group, bFGF group and NGF+bFGF group. Sciatic nerve function index (SFI), nerve conduction velocity, regeneration axon counting were detected in the 3 rd, 6 th, 9 th, 12 th week postinjury. RESULTS:The end of SFI, nerve conduction velocity, regeneration axon counting in NGF+bFGF group were higher than those in the other groups. CONCLUSION:The co effect of NGF and bFGF on sciatic nerve is better than the effect of NGF or bFGF alone.

THE EFFECT OF BASIC FIBROBLAST GROWTH FACTOR SLOW-RELEASE MICROCAPSULES ON ANGIOGENESIS IN INFARCTED RABBIT MYOCARDIUM

Objectives To observe the effect of basic fibroblast growth factor (bFGF) slow-release microcapsules on angiogenesis in infarcted myocardial regions. Methods.Myocardial infarction was induced in 24 New Zealand rabbits by ligating the root of left anterior descending coronary artery.Group Ⅰ(n=8) served as control, group Ⅱ(n=8) as a blank microcapsule group, group Ⅲ(n=8, each microcapsule contains 1μg bFGF) as micrpcapsule group.In group Ⅱ and Ⅲ, 5 blank microcapsules or bFGF slow-release microcapsules were implanted into myocardium underneath the epicardium between the left anterior descending coronary artery and left circumflex artery.Infarct size was evaluated by infarcted weight/left ventricle weight ratio and angiogenesis was evaluated by immunohistochemical examinations 5 weeks later. [WT5”BX] Results.As compared with group Ⅰ and Ⅱ, rabbits treated with bFGF slow-release microcapsules showed higher microvessel counts (group Ⅰ3775±450, group Ⅱ3837±498,vs.group Ⅲ 13550±481,P<0001) and less infarcted weight /left ventricle weight (group Ⅰ168%±04%,group Ⅱ167%±05%,vs.group Ⅲ 70%±02%,P<0001). Conclusions.Subepicardial administration of bFGF slow-release microcapsule in the infarcted rabbit model results in effective angiogenesis and reduction in infarct size.

Effects of 530 nm monochromatic light on basic fibroblast growth factor and transforming growth factor-β1 expression in Müller cells

AIM: To expose rat retinal Müller cells to 530 nm monochromatic light and investigate the influence of varying light illumination times on basic fibroblast growth factor(b FGF) and transforming growth factor-β1(TGF-β1) expression.·METHODS: Three groups of rat retinal Müller cells cultured in vitro under a 530 nm monochromatic light were divided into 6, 12 and 24 h experimental groups,while cells incubated under dark conditions served as the control group. The b FGF and TGF-β1 m RNA expression, protein levels and fluorescence intensity of the Müller cells were analyzed.·RESULTS: The b FGF m RNA expression and protein levels were significantly upregulated in Müller cells in all three experimental groups compared with the control group(P <0.05), while that of TGF-β1 was downregulated(P <0.05). Also, b FGF expression was positively correlated,but TGF-β1 expression was negatively correlated with illumination time. The largest changes for both cytokines were seen in the 24 h group. The changes in b FGF and TGF-β1 fluorescence intensity were highest in the 24 h group, and significant differences were observed among the experimental groups(P <0.05).·CONCLUSION: The expressions of b FGF and TGF-β1changed in a time-dependent manner in Müller cells exposed to 530 nm monochromatic light with 250 lx illumination intensity. Müller cells might play a role in the development of myopia by increasing b FGF expression or decreasing TGF-β1 expression. Changes in cytokine expression in retinal Müller cells may affect monochromatic light-induced myopia.

Effect of basic fibroblast growth factor on cat corneal endothelial cell proliferation

AIM: To investigate the function of basic fibroblast growth factor (bFGF) on cat corneal endothelial cells proliferation. METHODS: Cat corneal endothelial cells were primarily cultured, stimulated with bFGF for different period, the proliferation of cells was assayed by modified tertrozalium salt (MTT) method, and the morphologic changes were observed with inverted phase contrast microscope and transmission electron microscope. RESULTS: At 1, 3 and 5 days after bFGF was added to cat corneal endothelial cells, the result of MTT in 490nm showed significant difference than that in control group, and the difference was most significant in 10ng/mL group. CONCLUSION: bFGF can promote proliferation of cat corneal endothelial cells. 10ng/mL is the relatively most effective dose.

BASIC FIBROBLAST GROWTH FACTOR GENE TRANSFECTION TO ENHANCE THE REPAIR OF AVASCULAR NECROSIS OFTHE FEMORAL HEAD

Objective To explore a new method for the therapy of avascular necrosis of the femoral head. Method The recombinant plasmid pCD-rbFGF was mixed with collagen and was implanted in the necrotic femoral head. Expression of basic fibroblast growth factor (bFGF) was examined by RT-PCR and immunohistochemical method. Re-pair of the femoral head was observed by histological and histomorphometric analysis. Result Expression of bFGF was detected in the femoral head transfected with bFGF gene, indicating significant increase of angiogenesis 2 weeks after gene transfection and increased new bone formation 8 weeks after gene transfection on histom-orphometric analysis (P< 0.01). Conclusion Transfection of bFGF gene enhances bone tissue angiogenesis. Repair in osteonecrosis would be accelerated accordingly.

Expression of Basic Fibroblast Growth Factor in Rat Liver Fibrosis and Hepatic Stellate Cells

Summary:The expression of basic fibroblast growth factor (bFGF) in rat liver fibrosis and hepatic stellate cells (HSCs) and the relationship between the expression of bFGF and rat liver fibrogenesis were studied. Sixty male SD rats (230-260 g) were divided into 4 groups randomly (the 0 week group, 1 week group, 4 week group and 8 week group). Liver fibrosis was induced by subcutaneous injection of carbon tetrachloride. The sections of rats’ liver in each group were tested by Van-Gieson (V-G) staining and immunohistochemistry. The expression of bFGF mRNA was detected by reverse transcription polymerase chain reaction (RT-PCR). HSCs were isolated by the combined methods of collagenase IV perfusion and density gradient centrifugation. The expression of bFGF protein in cultured HSCs was detected by Western blot. Images of immunohistochemistry detection, agarose gel electrophoresis of RT-PCR and SDS-polyacrylamide gel electrophoresis of Western blot were analyzed semiquantitatively by image-analyzing system. The results were analyzed by statistics. The results showed that the fibers were gradually increased in the sections of rat liver with the prolongation of the model induction. At the end of the 8th weeks, liver fibrosis was formed. The expression of bFGF detected by immunohistochemistry showed a similar tendency of gradual increase. At the end of the 8th weeks, the bFGF expression could be observed in many regions in sections and the strongest expression was in interstitial cells including HSCs and some hepatocytes in regions around the portal area and central veins. Also there was moderate expression widely in extracellular matrix (ECM). In RT-PCR detection and Western blot detection of HSCs cultured in vitro, the similar tendency of gradual increase was evident either. It is suggested that bFGF is related with liver fibrosis of rats closely and may be a fibrogenesis factor of liver. bFGF possibly regulates liver fibrogenesis through regulating metabolism of extracellular matrix (ECM) by autocrine and paracrine stimulation.

Noggin versus basic fibroblast growth factor on the differentiation of human embryonic stem cells

The difference between Noggin and basic fibroblast growth factor for the neural precursor differen-tiation from human embryonic stem cells has not been studied. In this study, 100 μg/L Noggin or 20 μg/L basic fibroblast growth factor in serum-free neural induction medium was used to differen-tiate human embryonic stem cells H14 into neural precursors using monolayer differentiation. Two weeks after induction, significantly higher numbers of neural rosettes formed in the Noggin-induced group than the basic fibroblast growth factor-induced group, as detected by phase contrast micro-scope. Immunofluorescence staining revealed expression levels of Nestin, β-III Tubulin and Sox-1 were higher in the induced cells and reverse-transcription PCR showed induced cells expressed Nestin, Sox-1 and Neurofilament mRNA. Protein and mRNA expression in the Noggin-induced group was increased compared with the basic fibroblast growth factor-induced group. Noggin has a greater effect than basic fibroblast growth factor on the induction of human embryonic stem cell differentiation into neural precursors by monolayer differentiation, as Noggin accelerates and in-creases the differentiation of neural precursors.

Increased glycated basic fibroblast growth factor in diabetic skin reduces the cell viability and angiogenesis of human dermal microvascular endothelial cells

Objective To explore the glycation of basic fibroblast grow th factor( bFGF) in diabetic skin.Methods The abdom inal full-thickness skin tissues from 58 patients( 29 diabetic and 29 non-diabetic) aged 40 to69 years and granulation tissues from 15 patients( 8 diabetic and 7 non-diabetic) aged 50 to 59 were analyzed.The proportion of advanced glycation end products( AGEs)-b FG F intotal b FGF was measured with co-immunoprecipitation and the histological characteristics of wound skin were detected with hem atoxylin and eosin staining. The cell viability, apoptosis, and angiogenesis of human derm al m icrovascular endothelial cells( HDMECs) after exposure to AG Es-b FG For bFGF were measured with cell counting kit-8, flow cytom etry,and tube form ation assay, respectively. Results The proportion of AG Es-b FG F in total b FG F showed agedependent increaseboth in diabetic and non-diabetic skin. As com pared with non-diabetic skin, the constituent ratio in diabetic skin increased significantly in the equal age-group, and the same result could be obtained in granulation tissues from patients aged 50 to 59. The proportion of AG Es-b FG F in diabetic granulation was low er than that in diabetic skin from patients aged 50 to 59. H istological analysis show ed few er vessels in diabetic skin wound. In vitro, the viability and vascularization of H D M EC s were promoted by b FG F and inhibited after exposure to AGEs-bFGF for 7d. Conclusion The present study indicates that one cause for im paired wound healing in diabetic skin could be the glycated b FG F and its changed angiogenic function.

Effects of leukemia inhibitory factor and basic fibroblast growth factor on free radicals and endogenous stem cell proliferation in a mouse model of cerebral infarction

The present study established a mouse model of cerebral infarction by middle cerebral artery occlusion,and monitored the effect of 25 μg/kg leukemia inhibitory factor and(or) basic fibroblast growth factor administration 2 hours after model establishment.Results showed that following administration,the number of endogenous neural stem cells in the infarct area significantly increased,malondialdehyde content in brain tissue homogenates significantly decreased,nitric oxide content,glutathione peroxidase and superoxide dismutase activity significantly elevated,and mouse motor function significantly improved as confirmed by the rotarod and bar grab tests.In particular,the effect of leukemia inhibitory factor in combination with basic fibroblast growth factor was the most significant.Results indicate that leukemia inhibitory factor and basic fibroblast growth factor can improve the microenvironment after cerebral infarction by altering free radical levels,improving the quantity of endogenous neural stem cells,and promoting neurological function of mice with cerebral infarction.

Basic Fibroblast Growth Factor and Fibroblast Growth Factor Receptor-1 in Human Meningiomas

The expression of basic fibroblast growth factor (bFGF) and fibroblast growth factor receptor-1 (FGFR-1) in human meningiomas and the relationships between their expression and the tumors’ histological features and angiogenesis were investigated by means of immunohistochemical technique. The expression of bFGF and FGFR-1 was detected by antibody of bFGF or FGFR-1. The tumors’ angiogenesis was evaluated by microvascular density (MVD) and, which was observed by use of CD34-antibody immunohistochemically. The results showed that there were varied degrees of the expression of bFGF and FGFR-1 proteins in meningiomas. The expression was correlated with the tumors’ histological characters and angiogenesis. It was concluded that bFGF and FGFR-1 might play important roles in meningiomas’ angiogenesis and proliferation. The expression positive rate of bFGF and FGFR-1 may provide an indication of evaluating the histological and malignant degree of the tumor.

Effects of basic fibroblast growth factor on beta-catenin protein and mRNA expression during the proliferation of endogenous neural stem cells following focal cerebral ischemia

BACKGROUND:The Wnt/β-catenin signaling pathway plays an important role in neural development.β-catenin is an important component of the Wnt/β-catenin signaling pathway.The Wnt signaling pathway has been shown to regulate the interaction of neural stem cells with the extracellular matrix. OBJECTIVE:To investigate the effects of basic fibroblast growth factor(bFGF) onβ-catenin protein and mRNA expression,and on hippocampal neural stem cell proliferation in a rat model of cerebral ischemia/reperfusion. DESIGN,TIME AND SETTING:A randomized,controlled,neurobiology experiment was performed in Shenyang Medical College between August 2006 and August 2008. MATERIALS:A total of 72 healthy male Wistar rats,aged 3 months,were used in this study. bFGF was provided by Beijing SL Pharmaceutical Co.,Ltd.,China. METHODS:Rats were randomly divided into 3 groups:sham-operated,ischemia/reperfusion,and bFGF-treated(n=24 per group).Focal cerebral ischemia/reperfusion was induced in rats from the ischemia/reperfusion group and the bFGF-treated group by 2 hour right middle cerebral artery occlusion and 2 hour restoration of blood flow using the suture method.The ischemia/reperfusion and bFGF-treated groups were intraperitoneally administered 500 IU/mL of bFGF,or the same volume of physiological saline,once a day at postoperative days 1-3,and once every 3 days thereafter.Simultaneously,the sham-operated group underwent experimental procedures identical to the ischemia/reperfusion and bFGF-treated groups,with the exception of ischemia/reperfusion induction and drug administration.At 2 hours,2,6,13,and 20 days after ischemia/reperfusion induction,50 mg/kg bromodeoxyuridine(BrdU) was administered to each group,twice daily,to label proliferating neural stem cells. MAIN OUTCOME MEASURES:The effects of bFGF on BrdU labeling,andβ-catenin mRNA and protein expression,in neural stem cells were examined by immunohistochemistry,Western blot, RT-PCR,and in situ hybridization techniques. RESULTS:In the sham-operated group,only a few BrdU-immunoreactive neural stem cells were found.In the ischemia/reperfusion group,BrdU-immunoreactive cells began to increase from 3 days after ischemia/reperfusion induction,reached a peak level at 7 days,and gradually reduced from 21 days.At 3,7,14,and 21 days after ischemia/reperfusion induction,the numbers of BrdU-immunoreactive cells were significantly greater in the bFGF-treated group than in the ischemia/reperfusion group.The sham-operated group exhibited slight expression ofβ-catenin andβ-catenin mRNA.In the ischemia/reperfusion group,the expression ofβ-catenin andβ-catenin mRNA gradually increased with reperfusion time,peaked at 14 days after reperfusion, and gradually decreased thereafter;by 21 days,the expression was markedly lower.Following bFGF injection,the expression of hippocampal BrdU,β-catenin,andβ-catenin mRNA had apparently increased in each group. CONCLUSION:bFGF promotes neural stem cell proliferation,and the expression ofβ-catenin andβ-catenin mRNA in the ischemic brain tissue.These findings indicate that bFGF promotion of neural stem cell proliferation may be mediated by Wnt/β-catenin signaling pathway.