Apical ectodermal ridge regulates three principal axes of the developing limb / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Understanding limb development not only gives insights into the outgrowth and differentiation of the limb, but also has clinical relevance. Limb development begins with two paired limb buds (forelimb and hindlimb buds), which are initially undifferentiated mesenchymal cells tipped with a thickening of the ectoderm, termed the apical ectodermal ridge (AER). As a transitional embryonic structure, the AER undergoes four stages and contributes to multiple axes of limb development through the coordination of signalling centres, feedback loops, and other cell activities by secretory signalling and the activation of gene expression. Within the scope of proximodistal patterning, it is understood that while fibroblast growth factors (FGFs) function sequentially over time as primary components of the AER signalling process, there is still no consensus on models that would explain proximodistal patterning itself. In anteroposterior patterning, the AER has a dual-direction regulation by which it promotes the sonic hedgehog (Shh) gene expression in the zone of polarizing activity (ZPA) for proliferation, and inhibits Shh expression in the anterior mesenchyme. In dorsoventral patterning, the AER activates Engrailed-1 (En1) expression, and thus represses Wnt family member 7a (Wnt7a) expression in the ventral ectoderm by the expression of Fgfs, Sp6/8, and bone morphogenetic protein (Bmp) genes. The AER also plays a vital role in shaping the individual digits, since levels of Fgf4/8 and Bmps expressed in the AER affect digit patterning by controlling apoptosis. In summary, the knowledge of crosstalk within AER among the three main axes is essential to understand limb growth and pattern formation, as the development of its areas proceeds simultaneously.

Dynamic expression of wnt and fibroblast growth factor ligands in cleft palate induced by retinoic acid / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To screen the wnt and fibroblast growth factor (FGF) ligands involved in palatogenesis and cleft palate, and to study the dynamic expression of them in the different stages of palatal development and cleft palate formation.</p><p><b>METHODS</b>Mouse model of retinoic acid (RA)-induced cleft palate was set up. At embryo day (ED) 14.5, the palatal tissues of RA-treated group and wild type were collected and prepared for gene-chip analysis. According to the gene-chip results, wnt3, wnt8a, fgf9 and fgf10 were selected and their expression level was detected at ED13.5-15.5 by using semi-quantitative reverse transcription-PCR (RT-PCR).</p><p><b>RESULTS</b>(1) Gene-chip analysis showed that in RA-induced cleft palate group wnt8a and fgf9 were down-regulated, wnt3 and fgf10 were up-regulated in conversely. (2)During the different stage of the control group palatogenesis, intense expression of wnt3, wnt8a, fgf9 and fgf10 were detected with a continuous dynamic pattern. (3)Compared with the control group, the expression level of wnt3, wnt8a, fgf9 and fgf10 in RA-induced cleft palate showed significant difference, respectively (P < 0.05).</p><p><b>CONCLUSION</b>wnt and FGF signaling molecules participate in the palatogenesis, and RA pathway may interact with wnt and FGF signaling pathway.</p>

Effects of dexamethasone and vitamin B12 on expression of fibroblast growth factor 10 and fibroblast growth factor receptor 2b during early palatogenesis / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To observe the alteration of fibroblast growth factor 10 (Fgf10) and fibroblast growth factor receptor 2 (Fgfr2b) signal in mouse embryonic palate after dexamethasone and vitamin B12 exposure.</p><p><b>METHODS</b>Dams were divided teratogenetic group, antagomistic group and control group and were respectively injected dexamethasone, dexamethasone and vitamin B12, and normal sodium. Dams were killed and fetus was collected at embryo 12.5 and 13.5 day. The expression of Fgf10 and Fgfr2b and mesenchymal cells proliferation of mouse embryonic by western blotting and BrdU assay were checked.</p><p><b>RESULTS</b>Fgf10 and Fgfr2b expression was down-regulated and mesenchymal cells proliferation was inhibited significantly after dexamethasone exposure. After vitamin B12 treatment, Fgf10 and Fgfr2b expression did not restore, but cells proliferation was recovered.</p><p><b>CONCLUSION</b>Dexamethasone and vitamin B12 affected the expression of Fgf10 and Fgfr2b of mouse embryonic palate and mesenchyme cells proliferation, but the change was disaccord.</p>

Proteomic study of the effect of recombinant hFGF-10 adenovirus on HaCat cells / 药学学报

This study is to investigate the effect of recombinant hFGF-10 adenovirus on the proteome of HaCat cells, and to speculate further the possible mechanism of the effect of hFGF-10 on HaCat cells via differentially expressed proteins identified. Two-dimensional gel electrophoresis (2-DE) combined with tandem time-of-flight mass spectrometry was applied to identify the differentially expressed protein spots on the 2-DE maps of the whole-cell proteins from Ad infected and rAd-hFGF-10 infected HaCat cells. The mRNA and protein levels of the differentially expressed proteins were confirmed with semi-quantitative RT-PCR and Western blotting. The results showed that the 2-DE maps with high resolution were obtained, and four selected differentially expressed proteins involved in cell apoptosis, cytoskeleton regulation and protein degradation were identified with MALDI-TOF/TOF. The mRNA and protein levels of one of the differentially expressed proteins, VDAC2, were up-regulated in HaCat cells infected with the recombinant hFGF-10 adenovirus. The differentially expressed protein, VDAC2, may be related to the bioactivities of hFGF-10.

Cloning, expression of human keratinocyte growth factor and its purification and identification / 生物工程学报

To clone KGF-2 gene, get hKGF-2 protein and detemine its activity. The cNDA of human KGF-2 was isolated from fetal lung by RT-PCR and cloned into pBV220 plasmid. The recombinant pBV220-hKGF-2 plasmid was transformed into E. coli (BL21), induced at 42 degrees C for the expression of hKGF-2. Recombinant human KGF-2 was purified from the ultrasonic-treated BL21 by heparin-Sepharose CL-6B treated column chromatography and cation exchange column chromatography. MTT method was used for the determination of its biological activity. SDS-PAGE showed that rhKGF-2 was expressed in E. coli BL21 as soluble protein of approximately 20kD. The rhKGF-2 protein can stimulate the proliferation of NIH3T3 cells significantly from 1 ng/mL to 10 ng/mL. HKGF-2 cDNA wasclned and highly expressed in E. coli BL21 and the purified rhKGF-2 showed the mitogenic activity on NIH3T3 cells.

The effect of recombinant human fibro blast growth factor-10 (HFGF-10) on the biosynthesis of ADN in guinea-pig liver cells primitive culture

Guinea-pig liver cells primitive culture completed with 1.5-50 ng/ml of hFGF-10 promote the biosynthesis of ADN in these cells. The concentration of 25 ng/ml produces the highest effect in primitive culture of guinea-pig liver cells, the stimuli effect to mytosis of hFGF-10 is equal with hFGF-1 and hFGF-7. Heparin did not manifest any influence to the effect of hFGF-10 on guinea-pig primitive cultured cells