[Objective]To evaluate the effect of massive bioactive bone substitute in repairing large animals bone defect and to know its degrading rate.[Method]The massive Polylevolactic acid?collage calcium phosphate(PLLA?cTCP) carrieres by rapid forming technology was making,and then compounding rhBMP-2 and carrieres in a ratio of 3mg rhBMP-2 to one carrier was compounded to prepare the massive bioactive bone substitutes for dogs bone defect.Then the massive bone substitutes were implanted into 2.0cm dogss radius defects in the experiment group,and the massive carriers were implanted into in the control group.The repairing effect was evaluated by radiography,histology and biomechanics,and the degrading rate of the substitues was calculated in an image analysis apparatus.[Result]Radiographically,in the experiment group,the defects were connected by callus in all dogs in 12 weeks postoperatively;in 24 weeks,the callus rebuilt well.But in the control group,there was no callus formed in 24 weeks postoperatively,and the defects were not repaired.Histologically,in 12 weeks postoperatively,the outer layer of the callus in the experiment groups was lamellar bone and the center were trabecular bone,myeloid tissue and partial degrading carrier;in 24 weeks,the lamellar bone was more compact,trabecular bone decreased,myeloid tissue increased,and the carrier degraded more.In the control group,in 12 weeks postoperatively,the fibrous tissue wrapped and infiltrated into carrier,at the same time,part of the carrier degraded;in 24 weeks,the carrier was divided up by fibrous tissue and degraded more.The degrading rate of the carder in 12 weeks in the experiment group was 43.2%,in the control group was 35.7%,in 24 weeks 58.4% and 45.4%.Biomechanics,in 24 weeks after postoperation,the radius strength in the experiment group was superior to that in the normal bone.[Conclusion]The massive bioactive bone substitutes have satisfactory repairing effect on the radius bone defects of the large animal,but its degrading rate needs improving.

Objective To explore the role of growth factor receptor-bound 2 (GRB2) in homo sapiens eukaryotic translation elongation factor 1 alpha 2 (EEF1A2) promoting the carcinogenesis and development of pancreatic cancer.Methods The human pancreatic cancer cell line SW1990 with low expression of EEF1A2,was transfected by Ad5/F35-EEF1A2 plasmid.The expression of EEF1A2 in human pancreatic cancer cell lines BxPC-3 cells with high expression of EEF1A2,was down-regulated by small interfering RNA (siRNA) interference.The expressions of GRB2 in SW1990 and BxPC-3 cells were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot before and after interference.The pancreatic cancer BxPC-3 cells were interfered by specific and efficient chemical synthesized siRNA fragment.The changes of BxPC-3 cell proliferation and migration were observed by methyl thiazoly tetrazolium (MTT) and Transwell assay before and after interference.The data were analyzed by one-way analysis of variance.Results After human pancreatic cancer cell line SW1990 was transfected with Ad5/F35-EEF1A2 plasmid,the expression of GRB2 at mRNA and protein level both increased significantly (F=5.67,6.39,both P＜0.05).After the expression of EEF1A2 was inhibited by EEF1A2-siRNA,the expression of GRB2 at mRNA and protein level both decreased significantly (F=6.59,4.69,both P＜0.05).After BxPC-3 cells was transfected with siRNA-GRB2 for 72 hours,the results of MTT assay indicated that the absorbance value was 1.22±0.14,compared with negative control group (1.42±0.15) and blank control group (1.39 ± 0.15) the difference was statistically significant (F =6.63,P＜ 0.05).The results of transwell assay showed that the migration ability of cells in siRNA-GRB2 group decreased.After 24 hours,the number of migrated cells was 24.10±4.25,compared with negative control group (54.72±5.24) and blank control group (51.26 ± 6.23) the difference was statistically significant (F=55.00,P＜ 0.05).Conclusion GRB2 is key molecule in EEF1A2 promoting the progenesis and development of pancreatic cancer.

Restriction site amplification polymorphism (RSAP) markers were employed to access the genetic diversity and relationship of 120 lilyturf germplasms from different geographical origins. Sixteen RSAP primer pairs generated 326 polymorphic bands, of which 318 (97.55%) were polymorphic. The value of polymorphism information content (PIC) ranged from 0.87 to 0.95 with an average of 0.92. These results indicated there was abundant genetic diversity among samples. The results of data analysis on 20 population showed that the value of percentage of polymorphic locus (PPL), Nei's gene diversity (H) and Shannon's information index (I) were 19.94%-85.58%, 0.082 6-0.210 7, 0.120 6-0.328 1 respectively. The most abundant genetic diversity was found in the O. japonicus population from Zhejiang and the least in the Liriope minor population. The genetic distance among 20 population was 0.024 6-0.286 8, of which the minimum genetic distance was 0.024 6 between population I and population 13 while the maximum 0.286 8 between population 5 and population 15. Coefficient of genetic differentiation among natural populations was 0.115 3 (Gst). And the gene differentiation contributed to 43.07% of the total genetic variation among populations and to 56.93% within populations. The total gene flow (Nm) was 0.660 9. UPMGA clustering analysis was basically similar to of the principle coordinate analysis (PCA). The 120 samples were classified into four major groups, which were basically corresponded with the genetic relationships based on morphological traits. The results of UPMGA and PCA were also consistent with geographical origins.
Amplified Fragment Length Polymorphism Analysis ; China ; Genetic Variation ; Liriope Plant ; classification ; genetics ; Phylogeny ; Polymorphism, Restriction Fragment Length

Drought stress exerts a considerable effect on growth, physiology and secondary metabolisms of the medicinal plants. It could inhabit the growth of the medicinal plants but promote secretion of secondary metabolites. Other researches indicated that the medicinal plants could depend on the ABA signaling pathway and secreting osmotic substances to resist the drought stress and reduce the damage by it. The article concludes the changes in growth, physiology, secondary metabolisms and response mechanisms of medicinal plants to drought stress that provides a theoretical basis for exploring the relationship between medicinal plants and drought stress.
Abscisic Acid ; metabolism ; Droughts ; Plant Proteins ; genetics ; metabolism ; Plants, Medicinal ; genetics ; growth & development ; metabolism ; Signal Transduction ; Water ; metabolism

Objective To investigate the effects of over-expression of homo sapiens eukaryotic translation elongation factor 1 alpha 2 (EEF1A2) on in vitro invasion and lung metastasis of human pancreatic cancer SW1990 cells.Methods Letivirus-mediated delivery of EEF1A2 was used to enhance the expression of EEF1A2 gene in human pancreatic cancer SW1990 cells,the SW1990 cells stably over-expressing EEF1A2 protein (SW1990/EEF1A2 cells) were obtained,and the parent SW1990 cells and SW1990/GFP cells were used as the control,and the expressions of EEF1 A2 mRNA and protein were determined by Real-time PCR and Western blotting.The invasion ability of cells was determined by Transwell assay.The lung metastasis model was established by injection of SW1990 cells into the tail vein.Whole lung tissues were harvested,and visible nodules on tung surface were counted macroscopically 8 weeks later.Results The EEF1 A2 mRNA expression of SW1990/EEF1A2 was 3.252 ± 0.344,which was significantly higher than those in SW1990/GFP cells (1.000 ±0.060) and SW1990 cells (0.944 ±0.041,t =2.255,2.305,P＜0.01) ; the EEF1A2 protein expression was 0.833 ± 0.050,which was significantly higher than those in SW1990/GFP cells (0.247 ± 0.035) and SW1990 cells (0.273± 0.041,t=0.572,0.559,P＜0.01).The ability of invasion of SW1990/EEF1A2 cells was (60 ±4) cells,which was sigmificantly higher than (33 ±4) cells in SW1990/GFP group and (26 ± 3) cells in SW1990 group (t =31.33,34.78,P ＜ 0.01).Furthernore,SW1990/EEF1 A2 cells had a much higher incidence of lung metastasis in nude mice than SW1990/GFP cells and SW1990 cells in vivo (100％ vs.20％,20％,P ＜ 0.05).Conclusions EEF1 A2 over-expression can obviously increase the in vitro invasion and lung metastasis of pancreatic cancer SW1990 cells.

Although children with differentiated thyroid cancer ( DTC) are in a high proportion of cure, due to the absence of the standard management of the evaluation, treatment and follow-up of children with thyroid neoplasia, the aggressive therapy including total thyroidectomy followed by radioactive iodine ( RAI) ablation with iodine-131 ( 131 I) could not be ignored. For the majority of children, total or near-total thyroidectomy is recommended. Central neck dissection is recommended for children with malignant cytology and clinical evidence of gross extrathyroidal invasion and/or loco-regional metastasis on pre-operative. Selective use of 131 I in children with DTC was based on pre-and post-operative staging. Multi-disciplinary collaborative management is needed to improve their outcomes and to minimize adverse effects as far as possible.

Objective:To establish a liquid chromatography-mass spectrometry(LC-MS) method for determining the concentration of amlodipine besylate in human plasma and to evaluate the bioequivalence of 2 kinds of amlodipine besylate tablets.Methods: Twenty healthy male volunteers were enrolled into a single crossover study.A single dose of the suspension equivalent to 10 mg amlodipine besylate or a reference preparation was given in a crossover way.The plasma concentrations of amlodipine besylate were determined by LC-MS method in the volunteers at different time points;the pharmacokinetic parameters and relative bioavailability were calculated and the bioequivalence of the 2 preparations were evaluated.Results: The pharmacokinetic parameters for experimental and the reference preparations were: C_max(6.21?1.88) vs(6.03?1.08) ng/ml;AUC_0-120(250.68?52.61) vs(246.14?52.11) ng h/ml;T_max(6.0?2.3) vs(6.1? 2.5) h;t_1/2(40.45?6.68) vs(43.74?9.05) h,respectively.The linear range of the present method was 0.1-20.0 ng/ml;the lowest detectable concentration of amlodipine besylate was 0.1 ng/ml.There was no significant difference in pharmacokinetic parameters between the 2 tablets.Conclusion: The present method is simple to use,fast,and accurate.The 2 preparations of amlodipine besylate are bioequivalent.

Objective To investigate the cell-fusion modes and contractive characteristics of myotubes formed by muscle satellite cells in vitro. Methods The muscle satellite cells in quadriceps femoris of six 2-day-old SD rats were isolated, cultured and identified by immunofluorescence; the cell-fusion modes and contractive characteristics of myotubes were observed and recorded under microscope. Results The positve cells expressing marker Pax7 were obtained by immunofluorescence, and the purity was 90.3%. Myotubes were fused by several cells, cell and myotube or myotube and myotube. There were myotubes of spontaneous contraction on the conditions of no acetylcholine, no electric stimulation or no contiguous cells. Conclusion The myotubes of spontaneous contraction are fused by muscle satellite cells through three cell-fusion modes in vitro.

Objective To investigate the dose-effect effects of th ree relating bone growth factors, dexamethasone, recombinant human basic fibro blastic growth factor (rhFGF) and recombinant human bone morphogenetic protein- 2 (rhBMP-2), on proliferation and differentiation of periosteal cells so as to provide experimental basis for their further application in bone tissue engineer ing. Methods Periosteal cells were isolated and cultured in vitro and then exposed to dexamethasone (10 -8 mol/L, 10 -7 mol/L and 10 -6 mol/L), rhFGF (50 ng/ml, 200 ng/ml and 500 ng/ml) and rhBMP-2 (50 n g/ml, 500 ng/ml and 1 000 ng/ml) respectively. At the 4th and 7th days respectiv ely, the culture stopped and the total protein and alkaline phosphatase (ALP) ac tivities were measured. ResultsDexamethasone at concentratio n of 10 -6 mol/L significantly inhibited protein synthesis without obvious effects on ALP expression. The rhFGF at various concentrations significantly pro moted cell proliferation but inhibited ALP activity. The rhBMP-2 at various con centrations exerted insignificant effect on cell proliferation. In comparison, A LP expression was significantly enhanced by treatment of rhBMP-2 at concentrati on of 500 ng/ml and 1 000 ng/ml ( P

Objective:To detect the change of composition of immune cells in the spleen of miR-126 knockdown(miR-126KD) mice and preliminarily explore its significance .Methods: The expression level of miR-126 in spleens of miR-126KD mice was deter-mined by Realtime PCR.And the total number of splenocytes was calculated.The pathologic morphology change of the spleen was observed by HE staining.And the changes on proportion of DCs ,macrophages cells ,γδ T cells and NK T cells,CD3+T cells and its subgroup ,as well as CD19+B cells in spleens of miR-126KD mice were analyzed by Flow cytometry and calculated respectively.The level of phosphorylated AKt and NF-κB was analyzed by Western blot assay.Results:Compared with those of WT mice ,the expression level of miR-126 decreased obviously ( P<0.05 ) and the total number of cells increased obviously in spleen of miR-126 KD mice ( P<0.05).Moreover,the pathologic morphology of miR-126KD mice was significantly changed.The proportion and number of NK T cells in the inherent cells were significantly increased ( P<0.05 ) , but the proportion of Mφcells were significantly decreased ( P<0.05 ) . Meanwhile,the proportion and number of CD3+T cells and CD4+T cells in the adaptive immune cells were significantly increased (P<0.05),while the total number of CD19+B cells were significantly decreased (P<0.05).Last,the level of phosphorylation Akt and NF-κB increased obviously in spleen of miRNA-126 knockdown mice.Conclusion: Change obviously on the composition of immune cells subsets in the spleen of miRNA-126 knockdown mice ,which it may be related to the altered level of phosphorylated AKt and NF-κB and provides a preliminary experimental basis for the further exploring the roles of miR -126 in immune response.