In vitro and in vivo evaluations of nano-hydroxyapatite/polyamide 66/yttria-stabilized zirconia as a novel bioactive material for bone screws: Biocompatibility and bioactivity.

Zirconia and its derivatives have been receiving increased levels of attention with regard to their potential application in bone tissue engineering. These materials are of particular interest because of their excellent characteristics, such as superior biological and mechanical properties. In this study, yttria-stabilized tetragonal zirconia (YTZ)-reinforced nanohydroxyapatite/polyamide 66 (nHA/PA66) bone screws were prepared. The biocompatibility and bioactivity of nHA/PA66/YTZ were evaluated in vitro using MC3T3-E1 cells. Biocompatibility and bioactivity experiments (cell counting kit-8 tests, cell immunofluorescence analysis, and polymerase chain reaction) showed that nHA/PA66/YTZ could facilitate the biological functions of MC3T3-E1 cells. The attachment, proliferation, spreading, and expression of genes associated with osteogenesis (collagen 1, osteopontin, and osteocalcin) in cells cultured with the nHA/PA66/YTZ composite were all superior compared with the control groups (P < 0.05). In addition, nHA/PA66/YTZ bone screws were implanted into the femoral condyles of rabbits, and titanium screws were employed as a control group; postoperative histology and blood analysis revealed no obvious damage to the liver, kidneys, or any other major organs in either of the experimental groups. Moreover, nHA/PA66/YTZ screws resulted in significantly better bone-implant contact interfaces and enhanced formation of trabecular bone (P < 0.05); these characteristics were markedly better than those in the group that received titanium screws. These observations indicate that YTZ-reinforced nHA/PA66 composites have significant potential for applications in bone tissue engineering.

[Analysis of variance of repeated data measured by water maze with SPSS].

OBJECTIVE: To introduce the method of analyzing repeated data measured by water maze with SPSS 11.0, and offer a reference statistical method to clinical and basic medicine researchers who take the design of repeated measures. METHODS: Using repeated measures and multivariate analysis of variance (ANOVA) process of the general linear model in SPSS and giving comparison among different groups and different measure time pairwise. RESULTS: Firstly, Mauchly's test of sphericity should be used to judge whether there were relations among the repeatedly measured data. If any (P

[Effect of TX0201 on expression of the apoptosis signal transduction molecule caspase-3 and apoptosis associated genes bcl-2 and bax mRNA in brain tissue of rat analogue model of Alzheimer's disease].

OBJECTIVE: To study the mechanism of TX0201, an effective fraction extracted from Tiaoxin recipe in regulating apoptosis associated genes in brain tissue of rat analogue model of Alzheimer's disease (AD) induced by beta-amyloid protein 25-35 (Abeta 25-35). METHODS: The model of AD was induced by bilateral amygdala injection of Abeta 25-35 to study the spatial memory capacity using Morris water maze test, and by means of RT-PCR and immunohistochemistry assay, the expressions of beta-amyloid precursor protein (APP), apoptosis correlative genes (bcl-2, bax), and apoptosis signal transduction molecule (Caspase-3) in the brain, and the effect of TX0201 on expressions of these genes were examined. RESULTS: In AD model group, the spatial capacity was damaged significantly. Caspase-3 and the expression of APP mRNA and bax/bcl-2 mRNA were increased in the cortex and hippocampus; TX0201 ameliorated all the pathologic changes mentioned above. CONCLUSION: TX0201 could improve the oriented learning and memory capacity in AD rats by decreasing bax/bcl-2 and down-regulating Caspase-3 to reduce neurocyte apoptosis, suggesting that effective regulation of neuron apoptosis associated genes may be one of the mechanisms of TX0201.

[Effects of recipes for replenishing qi and activating blood on senescence related gene expressions in the liver of aging rats].

OBJECTIVE: To observe the effects of recipes for replenishing qi and activating blood on p16, p21, proliferating cell nuclear antigen (PCNA), cyclin D1 and cyclin E gene expressions in the liver of aging rats. METHODS: A recipe for replenishing qi and a recipe for activating blood were administered to aging rats respectively, and the effects of the above recipes on the expressions of senescence related genes (p16, p21, PCNA, cyclin D1 and cyclin E) were examined by RT-PCR and Western blotting methods. RESULTS: The expressions of p16, p21 and cyclin D1 mRNAs and proteins in the liver of the untreated aging rats were up-regulated, while the expressions of PCNA and cyclin E mRNAs and proteins decreased. As compared with the untreated aging rats, both recipes could down-regulate the expressions of cyclin D1 mRNA and protein and up-regulate the expressions of cyclin E mRNA and protein, but had no obvious effects on the expressions of mRNAs and proteins of p16, p21 and PCNA. CONCLUSION: Recipes for replenishing qi and activating blood can improve the liver cell proliferation of aging rats via down-regulating the expressions of cyclin D1 mRNA and protein and up-regulating the expressions of cyclin E mRNA and protein.

[Effect of recipes replenishing qi and activating blood on cell proliferation and apoptosis in the liver of aging rats].

OBJECTIVE: To observe the effect of recipes replenishing qi and activating blood on cell proliferation and apoptosis in the liver of natural aging rats. METHODS: Natural aging rats were under administration of recipes replenishing qi or activating blood for 4 months. The liver of the rats was prepared into cell suspension for determination of cell proliferation and apoptosis with PI-staining and flow cytometer. RESULTS: (1) Compared with those of the young rats, the cells in G(0)-G1 phase in the liver tissue of aging rats were increased (P< 0.01), and apoptosis cells were increased (P< 0.01), while the cells in S and G2-M phases were decreased (P< 0.01). (2) Compared with those of the aging rats, the cells in G(0)-G1 phase in the liver tissue of aging rats administered recipes replenishing qi or activating blood were decreased (P< 0.01), and it was more obvious in activating blood group than in replenishing qi group (P< 0.01); the cells in S and G2-M phases were increased (P< 0.01) and there was no significant difference between the activating blood group and the replenishing qi group (P> 0.05). (3) The apoptosis cells in replenishing qi or activating blood group were decreased significantly (P< 0.01), and the effect of replenishing qi was better than that of activating blood (P< 0.01). CONCLUSION: (1) Cell proliferation is decreased and apoptosis is increased in the liver tissue of natural aging rats. (2) Recipes replenishing qi or activating blood can accelerate cell proliferation in the liver tissue of natural aging rats, and the effect of activating blood was slightly stronger than that of replenishing qi. (3) Recipes replenishing qi or activating blood can inhibit cell apoptosis in the liver tissue of natural aging rats, and the effect of replenishing qi was better than that of activating blood.

[Effect of heart benefiting recipe in controlling IL-1beta, IL-6 and APPmRNA expression in brain of beta-amyloid protein induced rat model of dementia].

OBJECTIVE: To investigate the neuro-immune regulatory mechanism of Heart Benefiting recipe (HBR), an effective recipe for treatment of Alzheimer's disease (AD). METHODS: Using immunohistochemical and RT-PCR methods, the neuro-immunological pathological changes in the AD rat model induced by beta-amyloid protein (A beta1-40) via lateral cerebral ventricle injection, including mainly the glial fibrillary acidic protein expression and inflammatory cytokines IL-1beta, IL-6mRNA and beta-amyloid protein precursor (APPmRNA) gene expression were studied. And the effects of HBR on these parameters were observed. RESULTS: Deposition of A beta in cerebral tissue could induce activation of stellate glial cells and abnormal increased levels of inflammatory cytokines (IL-1beta and IL-6mRNA), also the elevation of APPmRNA level. HBR could effectively control the above-mentioned pathological changes. CONCLUSION: HBR could effectively control the inflammation and the A beta immune cascade reaction in brain of AD patients, it is one of the important therapeutic mechanisms of the recipe.

[Comparative study on the regulatory effects on senescence related cell cycle gene expression by TCM principles of tonifying shen, invigorating pi benefiting qi, and activating blood circulation].

OBJECTIVE: To compare the effects of TCM therapeutic principles of tonifying Shen (TS), benefiting Qi (BQ), invigorating Pi (IP) and activating blood circulation (ABC) herbs in regulating the gene expression in senescence related cell cycle. METHODS: Drug sera containing TCM herbs of the above-mentioned principles were used to treat the aged human diploid fibroblast cell line 2BS. The effect of TCM on the senescence related cell cycle and its related gene expression (P16INK4, Cyclin D1 and PCNA) were examined by means of cell proliferative doublings, flow cytometry, RT-PCR and Western blot analysis. RESULTS: TCM herbs of TS and BQ could improve the cell cycle, down-regulate the P16 and Cyclin D1 mRNA/protein expression, up-regulate PCNA mRNA/protein expression, while TCM herbs of IP and ABC showed insignificant effect on these indexes. CONCLUSION: TCM herbs of TS and BQ have effect in improving cell cycle, it may be achieved through promoting the P16 pathway of gene expression.

[Establishment of analogous oxidative damaged Alzheimer's disease rat model and effect of tiaoxin recipe on it].

OBJECTIVE: To establish a convenient, economical and practical analogous oxidative damaged Alzheimer's disease rat model (AD model) for exploring the effect of Tiaoxin Recipe (TXR) on the spatial memory capacity and beta-amyloid protein (A beta) deposition in the model. METHODS: The AD model was established by left ventricular injection of DHF-FeCl3-ADP. Spatial memory and learning capacity of the model rat was observed by Morris water maze method, A beta deposition in its cerebral cortex was observed by immunohistochemistry, and the effect of TXR was analysed. RESULTS: Compared with the normal group, the spatial memory capacity in the model group was obviously decreased, with A beta widely deposited in cortex, immunohistochemical examination showed that the number of A beta positive cells and their mean optic density significantly increased. TXR displayed significantly improving effect on the above-mentioned changes. CONCLUSION: The oxidative damaged model could not only express the clinical characteristics (short-term memory impairment), but also partially reflex the pathological changes (A beta deposition) of AD, is an economical and practical analogous AD model. TXR has the effects of improving spatial memory impairment and lowering A beta deposition in the AD model rats.