Effect of lentivirus encoding acidic fibroblast growth factor on cycle and proliferation of adipose-derived stem cells / 中华医学美学美容杂志

Objective To study the effect of the lentivirus encoding acidic fibroblast growth factor transfecting human adipose-derived stem cells (ADSCs) on the cell cycle and proliferation of ADSCs.Methods ADSCs were isolated and extracted by enzymatic digestion from the liposuction aspirate.ADSCs were cultured,identified and osteogenic induced reagent was used to induce differentiation of ADSCs towards bone cells.To obtain lentivirus encoding FGF-1,the plasmid PWPXLd FGF-1 was co-transfected with plasmid psPAX2,pMD2.G in 293T cells.ADSCs were infected with lentivirus encoding FGF-1.Expression of green fluorescent protein (GFP) in infected FGF-1 was observed by fluorescence microscope and expression of FGF-1 in ADSCs was verified by Western blot analysis.Flow cytometry was used to detect the cell cycle of ADSCs infected with lentivirus encoding FGF-1.EDU assay was performed to examine cell viability.Results Lentivirus encoding FGF-1 was obtained.After ADSCs being infected green fluorescence was found in about 70％ ADSCs,and overexpression of FGF-1 protein was detected in infceted cells by Western blot analysis.The percentage of G2/M phase cells was significantly increased compared with the control group,and the proliferation of ADSCs infected with lentivirus encoding FGF-1 was promoted as compared with the control group.Conclusions FGF-1 can enhance G2/M phase of ADSCs and promote the proliferation of ADSCs.

Alleviation of aortic vascular dysfunction in STZ/HFD-induced type 2 diabetic rats by nFGF1 / 中国病理生理杂志

AIM:To investigate the protective effect of non-mitogenic fibroblast growth factor 1 (nFGF1) on the aortic vascular function in streptozotocin (STZ)/high-fat diet (HFD)-induced type 2 diabetic rats and its underlying mechanisms. METHODS:Five-week-old male SD rats (n=30) were randomly divided into 3 groups (n=10 in each group),including normal control group,type 2 diabetic group and nFGF1 treatment group(type 2 diabetic rats were intra-peritoneally injected with 0.5 mg/kg nFGF1 every other day for 4 weeks). After the rats were sacrificed, blood glucose, cholesterol and triglyceride levels,aorta diastolic function and superoxide dismutase(SOD) level in the aorta of each group were measured. Besides,the protein levels of cyclooxygenase-2(COX-2),phosphorylated extracellular signal-regulated ki-nase (p-ERK) and endothelial nitric oxide synthase (eNOS) in the aorta were determined by Western blot. RESULTS:nFGF1 markedly lowered blood glucose, cholesterol and triglyceride levels, enhanced aorta SOD activity and upregulated protein level of eNOS in the type 2 diabetic rats. Furthermore,the increased protein levels of COX-2 and p-ERK in the type 2 diabetic rats were largely abrogated by nFGF1. CONCLUSION:nFGF1 effectively attenuates aortic vascular dysfunction in the type 2 diabetic rats,which may be associated with decreasing blood glucose,cholesterol and triglyceride levels,re-ducing inflammation and oxidative stress response,and activating eNOS signaling pathway.

Therapeutic effect of combined use of FGF1-loaded nano-liposomes and ultrasound-targeted microbubble destruction technique on treating rats with experimental diabetic cardiomyopathy / 中华心血管病杂志

Objective@#The therapeutic effect of acid fibroblast growth factor 1(FGF1) on rats with diabetic cardiomyopathy (DCM) was evaluated by using nano-liposomes combined with ultrasound-targeted microbubble destruction technique (UTMD).@*Methods@#The FGF1-loaded nano-liposomes were prepared by water-in-water emulsion method combined with lyophilization technique.TypeⅠdiabetes model was induced by intraperitoneal injection of streptozotocin (STZ, 70 mg/kg) in 60 male SD rats.Sixteen weeks later, diabetic rats were randomly divided into: placebo group (saline treatment), FGF1 group, FGF1-loaded nano-liposomes group, and FGF1-loaded nano-liposomes plus UTMD group (n=15 each). After two weeks of intervention followed by 2 weeks intervention stop, all rats underwent cardiac catheterization, and the left ventricular end-systolic pressure (LVESP), left ventricular end-diastolic pressure (LVEDP) and the maximal increase/decrease rate of left ventricular pressure (LV±dp/dtmax) were measured.Then, the rats were sacrificed and myocardial tissue were obtained for Masson trichrome staining, TUNEL apoptotic staining and CD31 immunohistochemistry staining to quantify myocardial collagen fraction (CVF), cardiac myocyte apoptotic index and myocardial microvascular density (MVD).@*Results@#(1)Scanning electron microscope results revealed good morphology and FGF1 encapsulation efficiency (84.3±2.8)% with high stability and dispensability of FGF1 loaded nano-liposomes.(2)The hemodynamic evaluation showed that LVESP, LV + dp/dtmax and LV -dp/dtmax were all significantly higher, while LVEDP was significantly lower in the FGF1-loaded nano-liposome+ UTMD group than in DCM group, FGF1 solution group, and FGF1 nano-liposome group(all P<0.05). (3)The Masson trichrome staining demonstrated that CVF was significantly higher in all DCM groups than in control group and was significantly lower in the FGF1-loaded nano-liposome+ UTMD group than in DCM group, FGF1 solution group, and FGF1 nano-liposome group (all P<0.05). (4)The CD31 immunohistochemical staining results showed that MVD was significantly lower in all DCM groups than in control group and was significantly higher in the FGF1-loaded nano-liposome+ UTMD group than in DCM group, FGF1 solution group, and FGF1 nano-liposome group (all P<0.05). (5)The TUNEL results showed that apoptotic index was significantly higher in all DCM groups than in control group and was significantly lower in the FGF1-loaded nano-liposome + UTMD group than in DCM group, FGF1 solution group, and FGF1 nano-liposome group (all P<0.05).@*Conclusion@#FGF1 nano-liposomes combining with UTMD technique can significantly improve cardiac functions and attenuate myocardial CVF and apoptosis and enhance myocardial MVD in DCM rats.

Effect of fibroblast growth factor 1 on the proliferating cell nuclear antigen expression in submandibular gland of diabetic mice / 中华口腔医学杂志

Objective@#To examine the proliferating cell nuclear antigen (PCNA) expression in submandibular gland of diabetic mice and to investigate the influence of fibroblast growth factor 1 (FGF-1) on PCNA expression and its possible mechanism.@*Methods@#Sixteen db/db diabetic male mice were randomly divided into diabetic group and diabetic-FGF-1 group (n=8). Eight age-matched db/m mice served as a control group. After FGF-1 was administered intraperitoneally to diabetic-FGF-1 group continuously for 16 weeks, blood glucose and body weight of each mouse in the three groups were detected at 0, 4, 8, 12, 16 weeks. Then the flow rate of saliva in three groups was compared at 0, 8, 16 weeks. At 16 week, bilateral submandibular glands were resected. Then HE staining was performed to observe the histological morphology of submandibular gland and PCNA expression was examined by immunohistochemical staining.@*Results@#Four weeks after administration, the blood glucose in diabetic-FGF-1 group decreased markedly, close to the control group (P>0.05). Weight loss in diabetic-FGF-1 group was noticeable at 8 weeks after administration, but still higher than that in the control group (P<0.05). The flow rate of saliva in diabetic-FGF-1 group increased gradually after administration, which was higher at 8, 16 weeks ([260.1±43.3], [308.5±34.0] mg·min-1·kg-1) respectively than that in the diabetic group at the same time point ([181.8±37.5], [194.9±49.8] mg·min-1·kg-1) (P<0.05). Compared with the control group, submandibular glands in diabetic group significantly atrophied and the glandular atrophy in diabetic-FGF-1 group was alleviated. The submandibular gland index in the control group, diabetic group and diabetic-FGF-1 group were (7.45±0.63), (2.23±0.26), (3.97±0.15) mg/g, respectively (P<0.05). HE staining showed that the histological morphology of submandibular gland in diabetic-FGF-1 group was clearer, and acinar and ductal atrophy were less significant than diabetic group. Immunohistochemistry showed that the rate of PCNA-positive cells in the control group, diabetic group and diabetic-FGF-1 group were (45.23±7.78)%, (11.50±1.69)%, (36.98±6.53)% respectively (P<0.05).@*Conclusions@#FGF-1 can up-regulate the expression of PCNA in submandibular gland of diabetic mice. This effect may be one of the important mechanisms of FGF-1 reversing the structural atrophy and dysfunction of submandibular gland caused by diabetes mellitus.

Mechanisms underlying interleukin-22-induced expression of heparin-binding epidermal growth factor-like growth factor in HaCaT cells / 中华皮肤科杂志

Objective To investigate the mechanisms underlying intedeukin-22 (IL-22)-induced expression of heparin-binding epidermal growth factor-like growth factor (HB-EGF) in HaCaT cells.Methods Some HaCaT cells were divided into several inverention groups treated with IL-22 at concentrations of 12.5,25,50,100 μg/L,respectively and a control group treated with phosphate buffer saline (PBS).After 24-hour culture,total proteins were extracted from the HaCaT cells,and Western blot was performed to measure the expression of phosphorylated extracellular signalregulated kinase 1/2 (P-ERK1/2) in the mitogen-activated protein kinase (MAPK)-ERK1/2 pathway,as well as phosphorylated-JAK2 (P-JAK2) and phosphorylated-signal transducer and activator of transcription 3 (P-STAT3) in the JAK2/STAT3 pathway.In a blocking experiment,some HaCaT cells were divided into 4 groups to be treated with PBS,IL-22,PD98059 (an inhibitor of MAPK-ERK1/2) combined with IL-22 (PD98059 group),AG490 (an inhibitor of JAK2/STAT3) combined with IL-22 (AG490 group),respectively.After 24-hour treatment,total proteins and mRNAs were extracted from the HaCaT cells followed by Western blot and real-time quantitative reverse transcription-PCR for the measurement of protein and mRNA expressions of HB-EGF respectively.Statistical analysis was carried out with the software SPSS 16.0 by one-way analysis of variance (ANOVA) for intergroup comparisons and by Bonferroni's test for multiple comparisons.Results After treatment with IL-22 at the above 4 concentrations,the expressions of P-ERK1/2,P-JAK2 and P-STAT3 in HaCaT cells were all increased compared with the control group (all P ＜ 0.05).The protein and mRNA expression levels (expressed as the HB-EGF/β-actin ratio and 2-△△Cr respectively) of HB-EGF were both significantly decreased in the PD98059 group and AG490 group than in the IL-22 group (protein:0.183 ± 0.020 and 0.199 ± 0.011 vs.0.924 ± 0.032,F =37.700,36.400,respectively,both P ＜ 0.05; mRNA:1.034 ± 0.072 and 0.989 ± 0.038 vs.1.844 ± 0.135,F =11.271,13.429,respectively,both P ＜ 0.05).Conclusions IL-22 can activate the MAPK-ERK1/2 and JAK2/STAT3 signaling pathways in HaCaT cells,which may contribute to IL-22-induced expression of HB-EGF in HaCaT cells.

Impact of Modified Bazhen Decoction on Bone Marrow Fibroblast Growth Factor-1 in Chronic Aplastic Anemia Patients / 广州中医药大学学报

Objective To observe the effect of modified Bazhen Decoction ( MBD) on bone marrow fibroblast growth factor-1 ( FGF-1) in chronic aplastic anemia ( CAA) patients. Methods Sixty cases of CAA were randomly divided into treatment group and control group, 30 cases in each group. Both groups were given conventional western medicine such as oral use of Stanozolol and Ciclosporin, and additionally, the treatment group received oral use of MBD. Six months constituted one treatment course for both groups. After treatment, the clinical efficiency in western medicine field and on Chinese medical syndromes was evaluated in both groups. The changes of peripheral hemogram, adverse reaction, and FGF-1 in the bone marrow of both groups were also monitored. Results ( 1) Clinical efficacy in western medicine field was 80.0% in the treatment group, and was 66.7%in the control group (P<0.05). (2) The total effective rate on Chinese medical syndromes was 100.0% in the treatment group, and was 83.3%in the control group (P<0.05). (3) After treatment, the scores of symptoms were decreased in both groups (P<0.05 or P<0.01 compared with those before treatment), and the decrease in the treatment group was superior to that in the control group (P<0.05). (4) Peripheral white blood cell (WBC) count, hemoglobin (HGB) content and platelet (PLT) count as well as bone marrow FGF-1 were increased in both groups after treatment (P<0.01 compared with those before treatment), and the increase of WBC, HGB, PLT and FGF-1 in the treatment group was superior to that in the control group (P<0.05). (5) In the aspect of the adverse reaction, the treatment group had 2 cases of hepatic fucntion injury, and one case of acne and hairiness; the control group had 2 cases of gum hypertrophy, 5 cases of hepatic function injury, and 4 cases of acne and hairiness. The control group had higher incidence of adverse reaction ( P<0.05). Conclusion MBD exerts certain therapeutic effect for CAA, and one of the possible mechanisms is related with the regulation of FGF-1 level.

Immunolocalization of FGF-2 and VEGF in rat periodontal ligament during experimental tooth movement

OBJECTIVE: This article aimed at identifying the expression of fibroblast growth factor-2 (FGF-2) and vascular endothelial growth factor (VEGF) in the tension and pressure areas of rat periodontal ligament, in different periods of experimental orthodontic tooth movement. METHODS: An orthodontic force of 0.5 N was applied to the upper right first molar of 18 male Wistar rats for periods of 3 (group I), 7 (group II) and 14 days (group III). The counter-side first molar was used as a control. The animals were euthanized at the aforementioned time periods, and their maxillary bone was removed and fixed. After demineralization, the specimens were histologically processed and embedded in paraffin. FGF-2 and VEGF expressions were studied through immunohistochemistry and morphological analysis. RESULTS: The experimental side showed a higher expression of both FGF-2 and VEGF in all groups, when compared with the control side (P < 0.05). Statistically significant differences were also found between the tension and pressure areas in the experimental side. CONCLUSION: Both FGF-2 and VEGF are expressed in rat periodontal tissue. Additionally, these growth factors are upregulated when orthodontic forces are applied, thereby suggesting that they play an important role in changes that occur in periodontal tissue during orthodontic movement. .

OBJETIVO: o objetivo desse estudo foi identificar a expressão do fator de crescimento de fibroblastos 2 (FGF-2) e do fator de crescimento vascular endotelial (VEGF) nos lados de tensão e pressão do ligamento periodontal de ratos, durante movimento ortodôntico experimental, em diferentes períodos de tempo. MÉTODOS: uma força ortodôntica de 0,5N foi aplicada no primeiro molar superior direito de 18 ratos Wistar machos, por períodos de 3 (grupo I), 7 (grupo II) e 14 dias (grupo III). O primeiro molar do lado oposto foi utilizado como controle. Os animais foram sacrificados nos períodos de tempo mencionados, sendo a arcada superior removida e fixada. Após a desmineralização, os espécimes foram processados histologicamente e embebidos em parafina. A expressão do FGF-2 e do VEGF foram estudadas por meio de análise imuno-histoquímica. RESULTADOS: o ligamento periodontal dos dentes submetidos à movimentação ortodôntica mostraram maior expressão tanto de FGF-2 quanto de VEGF, em todos os grupos experimentais, quando comparados com os dentes do lado controle (p < 0,05). Diferenças estatisticamente significativas entre os lados de tensão e pressão também foram encontradas nos dentes submetidos à movimentação ortodôntica. CONCLUSÕES: tanto o FGF-2 quanto o VEGF são expressos no tecido periodontal de ratos, e esses fatores de crescimento são aumentados quando forças ortodônticas são aplicadas, sugerindo que esses desempenham um papel importante na reorganização do periodonto durante o movimento ortodôntico. .

Continuous irrigation with recombinant human acid fibroblast growth factor combined with negative-pressure drainage for the treatment of refractory skin ulcer / 中华皮肤科杂志

Objective To evaluate the clinical efficacy of continuous irrigation with recombinant human acid fibroblast growth factor (rh-aFGF) combined with negative-pressure drainage in the treatment of refractory skin ulcer.Methods A clinical trial was performed from January 2010 to December 2012.Sixty patients with refractory skin ulcer were randomly and equally classified into three groups:group A receiving continuous irrigation with rhaFGF combined with negative-pressure drainage,group B receiving irrigation of raw surfaces with rh-aFGF followed by bandaging after debridement,group C receiving covering of raw surfaces with Vaseline gauze and absorbent gauze followed by bandaging after debridement.Dressings were changed once daily in every group.The treatment lasted 21 days.The efficacy of these regimens was evaluated on day 7,14 and 21.Results At the above time points for observation,both the granulation and epithelium tissues of ulcers grew well in group A,with a decrease in the volume of wounds and area of exposed deep tissues.There was a significant increase in the cure rate and reduction rate of wound volume in group A compared with the other two groups (all P ＜ 0.01).Conclusion Refractory dermal ulcer can be treated with continuous irrigation with rh-aFGF combined with negative-pressure drainage.

Resposta tecidual induzida por Biodentine™ e MTA Branco em subcutâneo de ratos / Tissue reaction induced by Biodentine™ and White MTA in rat subcutaneous

Biodentine™ é um biomaterial à base de silicato de cálcio produzido, segundo o fabricante, com o intuito de apresentar propriedades físico-químicas e biológicas superiores ao MTA. Nosso objetivo foi avaliar a resposta tecidual promovida por Biodentine™ (BDT) e MTA Angelus Branco em subcutâneo de ratos. Foram utilizados ratos adultos distribuídos em 3 grupos (n=20/grupo), segundo o material preenchendo os tubos de polietileno implantados no subcutâneo: BDT, MTA e GC (controle, tubos vazios). Após 7, 15, 30 e 60 dias, os implantes e os tecidos adjacentes foram processados para parafina. Cortes longitudinais das cápsulas adjacentes aos implantes foram corados com H&E, tricrômico de Masson, Picrosirius e submetidos ao Alcian Blue (AB) e reações imuno-histoquímicas para IL-6 e FGF-1. Obteve-se a densidade numérica de células inflamatórias (CI), de células imunomarcadas para IL-6 e FGF-1 e de mastócitos AB-positivos, além da porcentagem de colágeno birrefringente. Os dados foram submetidos à ANOVA e teste Tukey (p≤0,05). O número de CI e de células imunomarcadas para IL-6 e FGF-1 foi significantemente maior aos 7 dias em todos os grupos. Diferenças significantes no número de células IL-6-positivas não foram observadas entre BDT e MTA aos 30 e 60 dias; aos 60 dias, diferenças significantes no número de CI também não foram detectadas. O número de células FGF-1- positivas foi significantemente maior no grupo BDT em comparação ao grupo MTA, em todos os períodos. A densidade numérica de mastócitos e a porcentagem de colágeno aumentaram significantemente ao longo do tempo. Aos 60 dias, o número de mastócitos e o conteúdo de colágeno foram significantemente maiores nos grupos BDT e MTA, respectivamente. A redução significante do processo inflamatório, concomitante à redução na imunoexpressão para IL-6, indica que ambos os materiais são biocompatíveis. A acentuada imunoexpressão de FGF-1 aos 7 dias sugere que este fator deve ser responsável, pelo menos em parte, pela proliferação de fibroblastos e, consequentemente, estimula a formação de colágeno nas cápsulas. Os mastócitos devem ter um papel importante na remodelação das cápsulas, pois uma forte correlação foi detectada entre o aumento significante de mastócitos e de colágeno.

Biodentine™ is a new calcium silicate-based biomaterial which presents improved physicochemical and biological properties compared to MTA. The aim of this study was to evaluate the tissue reaction promoted by Biodentine™ (BDT) and MTA Angelus White in rat subcutaneous. Adult rats were distributed into 3 groups (n=20/group) according to the implanted materials: BDT, MTA or CG (Control group; empty tubes). After 7, 15, 30 and 60 days, the implants and adjacent tissues were fixed and embedded in paraffin. Longitudinal sections of the capsule adjacent to implants were stained with H&E, Masson's trichrome, Picrosirius and submitted to Alcian Blue (AB). Immunohistochemical reactions for IL-6 and FGF-1 were also performed. The number of inflammatory cells (IC), IL-6 and FGF-1 immunolabeled cells, AB-positive mast cells as well as birefringent collagen percentage were obtained. Data were statistically analyzed by ANOVA and Tukey test (p≤0.05). The number of IC and IL-6 and FGF-1 immunolabeled cells were significantly high at 7 days in all groups. At 30 and 60 days, significant differences in the number of IL-6-positive cells were not detected between BDT and MTA. At 60 days, statistical difference in the IC number was not observed between BDT and MTA groups. In all periods, the number of FGF-1-positive cells was significant higher in BDT group in comparison to MTA. The numerical density of mast cells and collagen percentage increased over time. At 60 days, mast cells number and collagen content were significantly high in BDT and MTA groups, respectively. A significant reduction of inflammatory process and IL-6 immunoexpression indicates that both materials are biocompatible. Intense FGF-1 immunoexpression at 7 days suggests that this factor may be responsible, at least in part, for fibroblast proliferation and subsequent collagen formation in the capsules. Since a strong correlation between mast cells number and collagen density was detected, it is conceivable to suggest that mast cells play a pivotal role in the capsules remodeling

Expressions of fibroblast growth factor-1, mitogen activated protein kinase-1 and CD34 in the lesions of patients with psoriasis vulgaris and their significance / 中华皮肤科杂志

Objective To estimate the relationship of fibroblast growth factor-1 (FGF-1),mitogen activated protein kinase-1 (MAPK-1) and CD34 expressions with clinicopathological findings in lesions of psoriasis vulgaris.Methods Skin specimens were collected from 30 patients with psoriasis vulgaris,whose general information and clinical findings were recorded.The protein expressions of FGF-1,MAPK-1 and CD34 in these specimens were detected by tissue chip technology and immunohistochemistry,and the mRNA expression of FGF-1 by reverse transcription-PCR.The correlations of FGF-1 protein expression with patients' gender,age,clinical stage,psoriasis area and severity index (PASI) score,angiogenesis and abnormal epidermal proliferation were statistically analyzed.Results The protein and mRNA expressions of FGF-1 were significantly higher in the lesions than in the control skin (both P ＜ 0.05).The protein expressions of CD34 and MAPK-1 were significantly increased,and positively associated with the protein expression of FGF-1 in psoriatic lesions.The up-regulation of FGF-1 protein was unrelated to the age or gender of patients,but associated with clinical stage and PASI score.The proportion of patients with increased FGF-1 expression was significantly larger in patients at active stage and with high PSAI score than in those at quiescent stage and with low PSAI score (P ＜ 0.05).Conclusions In psoriasis vulgaris,FGF-1 may participate in the abnormal epidermal proliferation and vascular dysplasia in dermal papilla,and the upregulation of FGF-1 appears to be associated with the progression of disease and aggravation of lesions.

A preliminary study of serum-deprivation-induced release of S100A13 and flbroblast growth factor 1 from thyroid cancer TT cell / 中华内分泌代谢杂志

Objective To investigate the effect of serum-deprivation on the changes of [ Ca2+ ] i and the protein release of S100A13 and fibroblast growth factor 1 ( FGF-1 ) in thyroid cancer TT cell,and to reveal the role of Ca2+in the protein release of S100Al3 and FGF-1.Methods The protein expressions of FGF-1 and S100A13 in TT cells under serum-deprivation were detected by Western blot.The released FGF-1 protein from TT cells in the supernatant fluid was detected by ELISA.Realtime dynamic examinations on the change of 1 h [ Ca2+ ] i in TT cells under serum-deprivation were detected by confocal laser scanning microscopy.Then,the effect of EGTA( 2.5 mmol/ L),BAPTA-AM (2.5 μmol/L)on distributions of the fluorescence of S100AI 3 and FGF-1 in TT cells under serumdeprivation for6 h were detected by indirect immunofluorescence.Results The expressions of FGF-1 and S100A13 in TT cells after serum-deprivation for4 h and 6 h were reduced( P＜0.05 or P＜0.01 ),but the released FGF-1 protein from TT cells in the supernatant fluid was elevated ( P＜0.05 or P＜0.01).Confocal laser scanning of Ca2+ imaging indicated that [ Ca2+ ] i of serum-deprivation TT cells maintained the relative stabilization within 23 win,but the rapid rise of [ Ca2+ ] i achieved peak value 1.6 μmol/L after 30 min,and remained stable for about 17 win,and thereafter 40 win slowly dropped to a low level From 40 win to 60 win the [ Ca2+ ] i was about 0.3-0.6 μ mol/L.The average [ Ca2+ ] i was higher than that in normal group,EGTA group,and BAPTA-AM group within 1 h.The protein expressions of S100A13 and FGF-1 did not drop obviously in EGTA group and BAPTA-AM group.Conlusion The release of S100A13 and FGF-1 from TT cell under serum-deprivation is possibly related with the change of [ Ca2+ ]i.Both Ca2+-chelating agents EGTA and BA PTA-AM are able to inhibit the rise of [ Ca2+ ] i and release of S100A 13 and FGF-1 from TT cells under serum-deprivation.

Effects of small hairpin RNA-mediated S100A13 gene inhibition on the release of fibroblast growth factor-1 in human thyroid cancer cells / 中华内分泌代谢杂志

Objective To investigate whether the release of fibroblast growth factor-1 ( FGF-1 ) was changed after inhibition of S100A13 gene (small hairpin RNA, shRNA)and serum-deprivation in human thyroid cancer cells (TT cells ). Methods The S100A13-shRNA pENTRTM/U6 entry vector was transfected into TT cells. The expression of S100A13 mRNA and protein was detected by immunoflurescence, real-time RT-PCR, and Western blot. Then TT cells were treated with S100A13 gene inhibition and serum-deprivation. The changes in release of FGF-1 were detected by indirect immunoflurescence, RT-PCR, and ELISA. Results S100A13 shRNA transfected TT cells (S100A13 RNAi cells)had a reduction of S100A13 gene and protein expression by 80%.Indirect immunofluorescence indicated FGF-1 was mostly localized in the cytoplasm and nucleus of TT cells in primary culture. When serum-deprivation stress was given to TT cells, FGF-1 in cytoplasm almost disappeared in the cells at 6 h. RT-PCR indicated that when serum-deprivation stress was given to TT cells the mRNA of FGF-1 was reduced. ELISA showed that with inhibition of S100A13, the release of FGF-1 was reduced (P＜0.05).Conclusion S100A13-shRNA pENTRTM/U6 entry vector transfected TT cells may inhibit the expression of S100A13 and reduce the release of FGF-1.

Correlation between the Promoter Polymorphism of Fibroblast Growth Factor 1 gene and Late-onset Alzheimer's Disease / 中国康复理论与实践

@#ObjectiveTo evaluate the correlation between the promoter polymorphism of Fibroblast Growth Factor 1 (FGF-1) and late-onset Alzheimer's disease (LOAD). MethodsClinic pathological data from 206 autopsies were analyzed, including 100 autopsy-confirmed LOAD patients and 106 age-matched non-demented controls. PCR-RFLP (Restriction fragment length polymorphism) approach was used to determine the genotype of the promoter polymorphism of FGF-1 gene. ResultsThe genotyping frequencies of the promoter polymorphism (-1385 A/G) were AA 20 (10%), GA 89 (43%), GG 97 (47%), respectively. There was significant (P=0.027) difference of genotyping frequencies between the cases and controls; GG genotype was positively associated with LOAD (odds ratio=2.02, 95%CI:1.16～3.52). ConclusionThe promoter polymorphism (-1385 A/G) of FGF-1 gene was associated with LOAD.

Protective effects of nmhaFGF on NRK52E cell apoptosis induced by H_2O_2 / 中国病理生理杂志

AIM:The present study was designed to establish H2O2-induced NRK52E cell apoptotic model and to investigate the effects and the mechanism of nmhaFGF on the NRK52E cell apoptosis induced by H2O2. METHODS: In vitro experiment, a apoptotic model of normal rat kidney epithelium (NRK52E) was made by MTT method, Hoechst33342 dyeing and flow cytosorting (FCR). NRK52E cells were cultured with different concentrations of nmhaFGF and haFGF for 24 h before H2O2 was added. The apoptotic rate was detected with FCR method. RESULTS: H2O2 at concentration of 0.4 mmol/L, the optimal condition to establish the apoptotic model, was used to treat NRK52E cells for 18h. Different doses of nmhaFGF (0.01, 0.03, 0.10, 0.30, 1.00 mg/L) reduced the apoptotic rates with the dose rising. However, the decreasing tends of apoptotic rates with dose increasing for haFGF was not so obvious. CONCLUSION: nmhaFGF protects the NRK52E cells against apoptosis. The mechanism might be connected with its non-mitogenic property.

Effects of human recombinant acidic fibroblast growth factor on cell proliferation and apoptosis in NIH3T3 cells / 中国病理生理杂志

AIM: Through studying the differences between wild-type acidic fibroblast growth factor (waFGF) with recombinant aFGF (raFGF), the biological effect of raFGF concerned with mitogenic activity was evaluated. METHODS: NIH3T3 cell line was used. Cell proliferation with MTT method was used to study the mitogenic activity. Flow cytometry was also used for detection of apoptosis, cell membrane permeability and mitochondria potential. The role of heparin sulfate (HS) on aFGF biological effect was studied at the same time in this research. RESULTS: The enhancement of raFGF on cell proliferation was significantly lower than that of waFGF. The restriction of raFGF on apoptosis and the enhancement of it on cell membrane permeability were all lower than those of waFGF significantly. The enhancement of raFGF on mitochondria potential was lower than that of waFGF significantly. The HS improved the biological effect of aFGF. CONCLUSION: The mitogenic activity of raFGF is lower than that of waFGF and raFGF has little effect on apoptosis.

Protection of maFGF against anoxia/reperfusion injury in isolated rat hearts / 中国病理生理杂志

AIM: To investigate the protective effects and mechanisms of modified acidic fibroblast growth factor (maFGF) in anoxic reperfusion of rat hearts. METHODS: Using Langendorff apparatus, we established the model of anoxia/reperfusion of isolated hearts to compare the protective effects of maFGF and acidic fibroblast growth factor (aFGF). The changes of left ventricle development pressure (LVDP) and maximal rates of rise of ventricular pressure(dp/dt_~max ), maximal rates of decline of ventricular pressure (dp/dt_~min ) were determined, changes of LDH and MDA levels,SOD activity in efflux from coronary artery were also detected at different time point. RESULTS: Pretreatment with maFGF and aFGF produced a similar protective effect on myocardium during anoxia /reperfusion, including promoting obviously heart functional recovery after myocardial anoxia/reperfusion and reversing changes of LDH, MDA contents and SOD activity induced by anoxic/reperfusion. CONCLUSION: maFGF has a protective effect on anoxia/reperfusion heart, and the mechanism of this effect may be related to suppression of lipid peroxidation.

The mitogenic activity decline of a haFGF mutant and its mechanism / 中国病理生理杂志

AIM: To observe the effects of the human acidic fibroblast growth factor mutant (mhaFGF), lacking 27 amino acids at N-terminal, on the proliferation and signal transduction of the hepatocarcinoma cells. METHODS: The hepatocarcinoma cells were treated with human acidic fibroblast growth factor (haFGF) and mhaFGF, respectively. The expression levels of the signal proteins, Grb2 and Erk1/2, in the hepatocarcinoma cells were detected by semi-quantitative Western-blotting after treated for 15 min. The mitogenic activity of both haFGF and mhaFGF was detected by MTT method and the cell cycle was analysed by flow cytometer (FCM) after treated for 48 h. RESULTS: The mitogenic activity and the ratio of G 1 and S phase cells in mhaFGF-treated cells were markedly lower than that of the haFGF, and close to that of the control group. The expression level of both Grb2 and Erk1/2 in the mhaFGF-treated cells were lower than those in the haFGF- treated cells. CONCLUSION: The decrease in the mitogenic activity of mhaFGF is probably associated to its down-regulating the expression of the signal molecular, MAPK-ERK1/2 and Grb2.