[Objective] The aim was to explore the optimal density and nitrogen rate of no-tilling and direct sowing rapeseeds in Chengdu plain. [Methods] Effects of in- teraction between density and nitrogen rate on the growth a...[Objective] The aim was to explore the optimal density and nitrogen rate of no-tilling and direct sowing rapeseeds in Chengdu plain. [Methods] Effects of in- teraction between density and nitrogen rate on the growth and yield of direct sowing rapeseed under no-tillage condition were investigated with Chuanyou 58 as materials and a split-plot experiment adopted. [Results] In Chengdu Plain, the yields of rape- seed changed from increasing to decreasing with increase of density and nitrogen rate. Both of density and nitrogen rate had significant effects on growth and yield of rapeseed and the latter overweighed in the effect. In addition, interaction of the two had negative effects on rapeseed yield. The yield of rapeseeds achieved the highest at 3 395.25 kg/hm^2 with interaction of density at 30.00×10^4 plant/hm^2 and nitrogen rate at 180.00 kg/hm^2; the theoretical maximal yield was 3 403.41 kg/hm^2 with interaction of density at 40.80×10^4 plants/hm^2 and nitrogen rate at 198.90 kg/hm^2. [Conclusion] In Chengdu Plain, the optimal density and nitrogen rate are 30.00×10^4-45.00×10^4 plant/hm^2 and 180.00-198.90 kg/hm^2, respectively.展开更多
An experiment was conducted to understand the growth inhibitory effects of aqueous extracts derived from Lantana camara L. (a globally recognized invasive alien weed) on six popular agricultural crops of Bangladesh....An experiment was conducted to understand the growth inhibitory effects of aqueous extracts derived from Lantana camara L. (a globally recognized invasive alien weed) on six popular agricultural crops of Bangladesh. The test was conducted in sterilized petridishes with a photoperiod of 24 hours and an average temperature of 29℃. The effect of different concentrations ofL. camara leaf extracts were recorded and compared with control (i.e., distil water). Result showed different concentrations of aqueous leaf extracts caused significant inhibitory effect on germination, root and shoot elongation and development of lateral roots of receptor crops. Bioassays also indicated that the inhibitory effect was proportional to the concentrations of the extracts and higher concentration had the stronger inhibitory effect whereas the lower concentration showed stimulatory effect in some cases. The inhibitory effect was much pronounced in root and lateral root development rather than shoot and germination.展开更多
The effects of auxin polar transport inhibitors, 9-hydroxy-fluorene-9-carboxylic acid (HFCA); 2, 3, 5-triiodobenzoic acid (TIBA) and trans-cinnamic acid (CA)on leaf pattern formation were investigated with shoots form...The effects of auxin polar transport inhibitors, 9-hydroxy-fluorene-9-carboxylic acid (HFCA); 2, 3, 5-triiodobenzoic acid (TIBA) and trans-cinnamic acid (CA)on leaf pattern formation were investigated with shoots formed from cultured leaf explants of tobacco and cultured pedicel explants of Orychophragmus violaceus, and the seedlings of tobacco and Brassica chinensis. Although the effective concentration varies with the inhibitors used, all of the inhibitors induced the formation of trumpet-shaped and/or fused leaves. The frequency of trumpet-shaped leaf formation was related to the concentration of inhibitors in the medium.Histological observation of tobacco seedlings showed that there was only one main vascular bundle and several minor vascular bundles in normal leaves of the control, but there were several vascular bundles of more or less the same size in the trumpet-shaped leaves of treated ones.These results indicated that auxin polar transport played an important role on bilateral symmetry of leaf growth.展开更多
AIM: To investigate the effects of thalidomide on angiogenesis, tumor growth and metastasis of hepatocellular carcinoma in nude mice.METHODS: Twenty-four nude mice were randomly divided into therapy group and control ...AIM: To investigate the effects of thalidomide on angiogenesis, tumor growth and metastasis of hepatocellular carcinoma in nude mice.METHODS: Twenty-four nude mice were randomly divided into therapy group and control group, 12 mice in each group. Thalidomide dissolved in 0.5% sodium carboxyl methyl cellulose (CMC) suspension was administered intraperitoneally once a day at the dose of 200 mg/kg in therapy group, and an equivalent volume of 0.5% CMC in control group. Mice were sacrificed on the 30th d, tumor size and weight and metastases in liver and lungs were measured. CD34 and VEGF mRNA in tumor tissue were detected by immunohistochemistry and semi-quantitative RT-PCR respectively and microvessel density (MVD) was counted. Serum concentrations of TNF-α and ALT and AFP were also tested.RESULTS: MVD and VEGF mRNA in therapy group were less than those in control group (31.08±16.23 vessels/HP vs 80.00±26.27 vessels/HP, 0.0538±0.0165 vs 0.7373±0.1297,respectively, P<0.05). No statistical difference was observed in tumor size and weight and metastases in liver and lungs.TNF-α was significantly lower in therapy group than in control group (28.64±4.64 ng/L vs42.69±6.99 ng/L, P<0.05). No statistical difference in ALT and AFP was observed between groups.CONCLUSION: Thalidomide can significantly inhibitangiogenesis and metastasis of hepatocellular carcinoma.Italso has inhibitory effects on circulating TNF-α.展开更多
To study the effect of the toxic secondary bile acid lithocholic acid (LCA) on the expression of fibroblast growth factor 19 (FGF19) in intestinal cells and to characterize the pregnane-X-receptor (PXR) response...To study the effect of the toxic secondary bile acid lithocholic acid (LCA) on the expression of fibroblast growth factor 19 (FGF19) in intestinal cells and to characterize the pregnane-X-receptor (PXR) response of the FGF19 promoter region. METHODS: The intestinal cell line LS174T was stimulated with various concentrations of chenodeoxy- cholic acid and lithocholic acid for several time points. FGF19 mRNA levels were determined with quantitative realtime RT-PCR. FGF19 deletion promoter constructs were generated and the LCA response was analzyed in reporter assays. Co-transfections with PXR and RXR were carried out to study FGF19 regulation by these factors, RESULTS: LCA and CDCA strongly up-regulate FGF19 mRNA expression in LS174T cells in a time and dose dependent manner. Using reporter gene assays with several deletion constructs we found that the LCA responsive element in the human FGF19 promoter maps to the proximal regulatory region containing two poten- tial binding sites for PXR. Overexpression of PXR and its dimerization partner retinoid X receptor (RXR) and stimulation with LCA or the potent PXR ligand rifampicin leads to a significant induction of FGF19 promoter activ- ity in intestinal cells. CONCLUSION: LCA induced feedback inhibition of bile acid synthesis in the liver is likely to be regulated by PXR inducing intestinal FGF19 expression.展开更多
Objective: We established a transplantation tumor model of human osteosarcoma in chick embryo, studied its morphological and biological characteristics, and observed its dynamic process of angiogenesis induction so th...Objective: We established a transplantation tumor model of human osteosarcoma in chick embryo, studied its morphological and biological characteristics, and observed its dynamic process of angiogenesis induction so that a simple and practical model can be provided for studying osteosarcoma. Methods: Human osteosarcoma cells at different concentrations were inoculated in chorioallantoic membrane (CAM) of chick embryos at different embryonic ages to observe the factors affecting the survival of the transplanted osteosarcoma in chick embryo, growth characteristics of the transplantation tumor, and the morphological characteristics and biological characteristics of the transplantation tumor. Results: The transplantation tumor model of human osteosarcoma in chick embryo was successfully established. It was found that the transplantation tumor was easy to grow and it showed strong angiogenesis-inducing effects. Under the light microscope, the transplantation tumor showed a similar tissue structure to human osteosarcoma. Conclusion: It is feasible to establish a transplantation tumor model of human osteosarcoma in chick embryo. The model can be easily duplicated with a simple operation, which provides a useful animal model for studying osteosarcoma.展开更多
AIM: To investigate the effect of (-)-epigallocatechin-3-gallate (EGCG) on growth of gastric cancer and its possible mechanism. METHODS: Heterotopic tumors were induced by subcutaneously injection of SGC-7901 ce...AIM: To investigate the effect of (-)-epigallocatechin-3-gallate (EGCG) on growth of gastric cancer and its possible mechanism. METHODS: Heterotopic tumors were induced by subcutaneously injection of SGC-7901 cells in nude mice. Tumor growth was measured by calipers in two dimensions. Tumor angiogenesis was determined with tumor microvessel density (MVD) by immunohistology. Vascular endothelial growth factor (VEGF) protein level and activation of signal transducer and activator of transcription 3 (Star3) were examined by Western blotting. VEGF mRNA expression was determined by RT-PCR and VEGF release in tumor culture medium by ELISA. VEGF-induced cell proliferation was studied by MTT assay, cell migration by gelatin modified Boyden chamber (Transwell) and in vitro angiogenesis by endothelial tube formation in Matrigel. RESULTS: Intraperitoneal injection of EGCG inhibited the growth of gastric cancer by 60.4%. MVD in tumor tissues treated with EGCG was markedly reduced. EGCG treatment reduced VEGF protein level in vitro and in vivo. Secretion and mRNA expression of VEGF in tumor cells were also suppressed by EGCG in a dose-dependent manner. This inhibitory effect was associated with reduced activation of Star3, but EGCG treatment did not change the total Star3 expression. EGCG also inhibited VEGF-induced endothelial cell proliferation, migration and tube formation. CONCLUSION: EGCG inhibits the growth of gastric cancer by reducing VEGF production and angiogenesis, and is a promising candidate for anti-angiogenic treatment of gastric cancer.展开更多
Controllable syntheses of different-sized gold nanoclusters are of great significance for their fundamental science and practical applications. In this work, we achieve the controllable and selective syntheses of Au7 ...Controllable syntheses of different-sized gold nanoclusters are of great significance for their fundamental science and practical applications. In this work, we achieve the controllable and selective syntheses of Au7 and Au13 clusters through adding HCl to the traditional Aull synthetic route at different reaction time. Time-dependent mass spectra and UV- Vis spectra were ernployed to monitor these two HCl-directed processes, and revealed the distinct roles of HCl as an etchant or a growth prornotor, respectively. Furthermore, parallel experiments on independent synthetic routes involving only non-chlorine H+ (acetic acid) or Cl (tetraethy larnrnoniurn chloride) instead of HCl were perforrned, which illustrated the main role of H^+-etching and Cl^ -assisted growth in HCl-directed cluster synthetic routes. We propose the HCl-etching is mainly achieved via the H+ action to break the Au(I)-PPh3 part of clusters, while the HCl-prornoted growth is realized via the attachment of Au-Cl species to the pre-forrned clusters.展开更多
The thermodynamics and the growth kinetics of synthesis of the SiC whiskers (SiC w ) from rice hulls are studied in this paper. The results show that the intimate contact of SiO 2 with C in the rice hulls resulted in ...The thermodynamics and the growth kinetics of synthesis of the SiC whiskers (SiC w ) from rice hulls are studied in this paper. The results show that the intimate contact of SiO 2 with C in the rice hulls resulted in the formation of SiC particle (SiC p ) at lower temperature, and the external ash of the hulls (w (SiO 2 )>98%) is the main silicon source for SiCw growth. The metallic composite catalyst increases the selectivity for SiCw growth and the reaction rate. The growth mechanism of the SiCw can be characterized as the VLS (vapour liquid solid) with the presence of the whisker forming catalyst: from SiC nucleation through enlargement and growing with the <1 1 1> crystallographic orientation in a certain diameter, then the SiC w is a complete single crystal of β SiC. The generation reaction of SiO is the rate determing step for synthesis of SiC w .展开更多
H. suaveolens (L.) Poit has been used in folk medicine revealing a wide pharmacological potential. This study aimed to evaluate the efficacy of the essential oil from 11. suaveolens leaves in inhibiting the growth o...H. suaveolens (L.) Poit has been used in folk medicine revealing a wide pharmacological potential. This study aimed to evaluate the efficacy of the essential oil from 11. suaveolens leaves in inhibiting the growth of A. flavus LM257 and synthesis of aflatoxins Bl and B2. The essential oil at 20, 40 and 80 laL/mL strongly reduced the biomass and spore germination ofA. flavus. A 100% inhibition was found at 40 μL/mL and 80μL/mL, while at 20 μL/mL the growth depressed over 50% for both tested parameters. The essential oil at all assayed concentrations totally inhibited the production of aflatoxin B1 and B2. From these results, the essential oil from 11. suaveolens leaves may be considered as an alternative and effective inhibitor ofA. flavus, in addition to offer some protective effect to the production of aflatoxins.展开更多
Objective: To explore chemical component changes of dog bone at different lengthening time and in different bone regions of interest, and to evaluate the mineralization during Ilizarov lengthening process. Methods: Th...Objective: To explore chemical component changes of dog bone at different lengthening time and in different bone regions of interest, and to evaluate the mineralization during Ilizarov lengthening process. Methods: The ash weight, the concentrations of calcium, phosphorus and the calcium/phosphorus ratio were measured at different intervals (2, 4, 6, 8, 12 weeks) since lengthening and the lengthened part was compared with a control area at each interval. Results: The ash weight, the concentrations of calcium and phosphorus in the lengthened area differed at all development time. The calcium/phosphorus (Ca/P) ratio in the lengthened region remained significantly lower than that in the control region up to 12 weeks after the lengthening. Conclusions: These results suggest that also other inorganic ions play an important role in the mineralization process and that they become relatively more important since 8 weeks after the lengthening.展开更多
基金Supported by National Science and Technology Support Program(2010BAD01B08)Sichuan Finance Gene Engineering Program(2011JYGC04-013)12th Five Year Breeding Project of Crops of Sichuan Province~~
文摘[Objective] The aim was to explore the optimal density and nitrogen rate of no-tilling and direct sowing rapeseeds in Chengdu plain. [Methods] Effects of in- teraction between density and nitrogen rate on the growth and yield of direct sowing rapeseed under no-tillage condition were investigated with Chuanyou 58 as materials and a split-plot experiment adopted. [Results] In Chengdu Plain, the yields of rape- seed changed from increasing to decreasing with increase of density and nitrogen rate. Both of density and nitrogen rate had significant effects on growth and yield of rapeseed and the latter overweighed in the effect. In addition, interaction of the two had negative effects on rapeseed yield. The yield of rapeseeds achieved the highest at 3 395.25 kg/hm^2 with interaction of density at 30.00×10^4 plant/hm^2 and nitrogen rate at 180.00 kg/hm^2; the theoretical maximal yield was 3 403.41 kg/hm^2 with interaction of density at 40.80×10^4 plants/hm^2 and nitrogen rate at 198.90 kg/hm^2. [Conclusion] In Chengdu Plain, the optimal density and nitrogen rate are 30.00×10^4-45.00×10^4 plant/hm^2 and 180.00-198.90 kg/hm^2, respectively.
文摘An experiment was conducted to understand the growth inhibitory effects of aqueous extracts derived from Lantana camara L. (a globally recognized invasive alien weed) on six popular agricultural crops of Bangladesh. The test was conducted in sterilized petridishes with a photoperiod of 24 hours and an average temperature of 29℃. The effect of different concentrations ofL. camara leaf extracts were recorded and compared with control (i.e., distil water). Result showed different concentrations of aqueous leaf extracts caused significant inhibitory effect on germination, root and shoot elongation and development of lateral roots of receptor crops. Bioassays also indicated that the inhibitory effect was proportional to the concentrations of the extracts and higher concentration had the stronger inhibitory effect whereas the lower concentration showed stimulatory effect in some cases. The inhibitory effect was much pronounced in root and lateral root development rather than shoot and germination.
文摘The effects of auxin polar transport inhibitors, 9-hydroxy-fluorene-9-carboxylic acid (HFCA); 2, 3, 5-triiodobenzoic acid (TIBA) and trans-cinnamic acid (CA)on leaf pattern formation were investigated with shoots formed from cultured leaf explants of tobacco and cultured pedicel explants of Orychophragmus violaceus, and the seedlings of tobacco and Brassica chinensis. Although the effective concentration varies with the inhibitors used, all of the inhibitors induced the formation of trumpet-shaped and/or fused leaves. The frequency of trumpet-shaped leaf formation was related to the concentration of inhibitors in the medium.Histological observation of tobacco seedlings showed that there was only one main vascular bundle and several minor vascular bundles in normal leaves of the control, but there were several vascular bundles of more or less the same size in the trumpet-shaped leaves of treated ones.These results indicated that auxin polar transport played an important role on bilateral symmetry of leaf growth.
文摘AIM: To investigate the effects of thalidomide on angiogenesis, tumor growth and metastasis of hepatocellular carcinoma in nude mice.METHODS: Twenty-four nude mice were randomly divided into therapy group and control group, 12 mice in each group. Thalidomide dissolved in 0.5% sodium carboxyl methyl cellulose (CMC) suspension was administered intraperitoneally once a day at the dose of 200 mg/kg in therapy group, and an equivalent volume of 0.5% CMC in control group. Mice were sacrificed on the 30th d, tumor size and weight and metastases in liver and lungs were measured. CD34 and VEGF mRNA in tumor tissue were detected by immunohistochemistry and semi-quantitative RT-PCR respectively and microvessel density (MVD) was counted. Serum concentrations of TNF-α and ALT and AFP were also tested.RESULTS: MVD and VEGF mRNA in therapy group were less than those in control group (31.08±16.23 vessels/HP vs 80.00±26.27 vessels/HP, 0.0538±0.0165 vs 0.7373±0.1297,respectively, P<0.05). No statistical difference was observed in tumor size and weight and metastases in liver and lungs.TNF-α was significantly lower in therapy group than in control group (28.64±4.64 ng/L vs42.69±6.99 ng/L, P<0.05). No statistical difference in ALT and AFP was observed between groups.CONCLUSION: Thalidomide can significantly inhibitangiogenesis and metastasis of hepatocellular carcinoma.Italso has inhibitory effects on circulating TNF-α.
文摘To study the effect of the toxic secondary bile acid lithocholic acid (LCA) on the expression of fibroblast growth factor 19 (FGF19) in intestinal cells and to characterize the pregnane-X-receptor (PXR) response of the FGF19 promoter region. METHODS: The intestinal cell line LS174T was stimulated with various concentrations of chenodeoxy- cholic acid and lithocholic acid for several time points. FGF19 mRNA levels were determined with quantitative realtime RT-PCR. FGF19 deletion promoter constructs were generated and the LCA response was analzyed in reporter assays. Co-transfections with PXR and RXR were carried out to study FGF19 regulation by these factors, RESULTS: LCA and CDCA strongly up-regulate FGF19 mRNA expression in LS174T cells in a time and dose dependent manner. Using reporter gene assays with several deletion constructs we found that the LCA responsive element in the human FGF19 promoter maps to the proximal regulatory region containing two poten- tial binding sites for PXR. Overexpression of PXR and its dimerization partner retinoid X receptor (RXR) and stimulation with LCA or the potent PXR ligand rifampicin leads to a significant induction of FGF19 promoter activ- ity in intestinal cells. CONCLUSION: LCA induced feedback inhibition of bile acid synthesis in the liver is likely to be regulated by PXR inducing intestinal FGF19 expression.
文摘Objective: We established a transplantation tumor model of human osteosarcoma in chick embryo, studied its morphological and biological characteristics, and observed its dynamic process of angiogenesis induction so that a simple and practical model can be provided for studying osteosarcoma. Methods: Human osteosarcoma cells at different concentrations were inoculated in chorioallantoic membrane (CAM) of chick embryos at different embryonic ages to observe the factors affecting the survival of the transplanted osteosarcoma in chick embryo, growth characteristics of the transplantation tumor, and the morphological characteristics and biological characteristics of the transplantation tumor. Results: The transplantation tumor model of human osteosarcoma in chick embryo was successfully established. It was found that the transplantation tumor was easy to grow and it showed strong angiogenesis-inducing effects. Under the light microscope, the transplantation tumor showed a similar tissue structure to human osteosarcoma. Conclusion: It is feasible to establish a transplantation tumor model of human osteosarcoma in chick embryo. The model can be easily duplicated with a simple operation, which provides a useful animal model for studying osteosarcoma.
基金Supported by the grants from the National Natural Science Foundation of China, No. 30571833the Natural Science Foundation of Guangdong Province, China, No. 05001785
文摘AIM: To investigate the effect of (-)-epigallocatechin-3-gallate (EGCG) on growth of gastric cancer and its possible mechanism. METHODS: Heterotopic tumors were induced by subcutaneously injection of SGC-7901 cells in nude mice. Tumor growth was measured by calipers in two dimensions. Tumor angiogenesis was determined with tumor microvessel density (MVD) by immunohistology. Vascular endothelial growth factor (VEGF) protein level and activation of signal transducer and activator of transcription 3 (Star3) were examined by Western blotting. VEGF mRNA expression was determined by RT-PCR and VEGF release in tumor culture medium by ELISA. VEGF-induced cell proliferation was studied by MTT assay, cell migration by gelatin modified Boyden chamber (Transwell) and in vitro angiogenesis by endothelial tube formation in Matrigel. RESULTS: Intraperitoneal injection of EGCG inhibited the growth of gastric cancer by 60.4%. MVD in tumor tissues treated with EGCG was markedly reduced. EGCG treatment reduced VEGF protein level in vitro and in vivo. Secretion and mRNA expression of VEGF in tumor cells were also suppressed by EGCG in a dose-dependent manner. This inhibitory effect was associated with reduced activation of Star3, but EGCG treatment did not change the total Star3 expression. EGCG also inhibited VEGF-induced endothelial cell proliferation, migration and tube formation. CONCLUSION: EGCG inhibits the growth of gastric cancer by reducing VEGF production and angiogenesis, and is a promising candidate for anti-angiogenic treatment of gastric cancer.
基金supported by the National Natural Science Foundation of China (No.11475176, No.U1632263, and No.21533007)the Foundation for Innovative Research Groups of the National Natural Science Foundation of China (No.11621063)
文摘Controllable syntheses of different-sized gold nanoclusters are of great significance for their fundamental science and practical applications. In this work, we achieve the controllable and selective syntheses of Au7 and Au13 clusters through adding HCl to the traditional Aull synthetic route at different reaction time. Time-dependent mass spectra and UV- Vis spectra were ernployed to monitor these two HCl-directed processes, and revealed the distinct roles of HCl as an etchant or a growth prornotor, respectively. Furthermore, parallel experiments on independent synthetic routes involving only non-chlorine H+ (acetic acid) or Cl (tetraethy larnrnoniurn chloride) instead of HCl were perforrned, which illustrated the main role of H^+-etching and Cl^ -assisted growth in HCl-directed cluster synthetic routes. We propose the HCl-etching is mainly achieved via the H+ action to break the Au(I)-PPh3 part of clusters, while the HCl-prornoted growth is realized via the attachment of Au-Cl species to the pre-forrned clusters.
文摘The thermodynamics and the growth kinetics of synthesis of the SiC whiskers (SiC w ) from rice hulls are studied in this paper. The results show that the intimate contact of SiO 2 with C in the rice hulls resulted in the formation of SiC particle (SiC p ) at lower temperature, and the external ash of the hulls (w (SiO 2 )>98%) is the main silicon source for SiCw growth. The metallic composite catalyst increases the selectivity for SiCw growth and the reaction rate. The growth mechanism of the SiCw can be characterized as the VLS (vapour liquid solid) with the presence of the whisker forming catalyst: from SiC nucleation through enlargement and growing with the <1 1 1> crystallographic orientation in a certain diameter, then the SiC w is a complete single crystal of β SiC. The generation reaction of SiO is the rate determing step for synthesis of SiC w .
文摘H. suaveolens (L.) Poit has been used in folk medicine revealing a wide pharmacological potential. This study aimed to evaluate the efficacy of the essential oil from 11. suaveolens leaves in inhibiting the growth of A. flavus LM257 and synthesis of aflatoxins Bl and B2. The essential oil at 20, 40 and 80 laL/mL strongly reduced the biomass and spore germination ofA. flavus. A 100% inhibition was found at 40 μL/mL and 80μL/mL, while at 20 μL/mL the growth depressed over 50% for both tested parameters. The essential oil at all assayed concentrations totally inhibited the production of aflatoxin B1 and B2. From these results, the essential oil from 11. suaveolens leaves may be considered as an alternative and effective inhibitor ofA. flavus, in addition to offer some protective effect to the production of aflatoxins.
文摘Objective: To explore chemical component changes of dog bone at different lengthening time and in different bone regions of interest, and to evaluate the mineralization during Ilizarov lengthening process. Methods: The ash weight, the concentrations of calcium, phosphorus and the calcium/phosphorus ratio were measured at different intervals (2, 4, 6, 8, 12 weeks) since lengthening and the lengthened part was compared with a control area at each interval. Results: The ash weight, the concentrations of calcium and phosphorus in the lengthened area differed at all development time. The calcium/phosphorus (Ca/P) ratio in the lengthened region remained significantly lower than that in the control region up to 12 weeks after the lengthening. Conclusions: These results suggest that also other inorganic ions play an important role in the mineralization process and that they become relatively more important since 8 weeks after the lengthening.