Objective To evaluate the biocompatibility and safety of a novel orthopedics materials-graded zirconia(ZrO2)hydroxyapatite(HA) composite biomaterials. Methods First, ultrafine powers of ZrO2 and HA powder were prepared by chemical precipitation method, then graded ZrO2-HA composite was synthesized by dry-laying and sintering method. After the physiological saline and culture medium extracts of the composite were prepared, four experiments were conducted as follows:① The mouse acute toxic test consists of 2 groups(n=10). The extracts were intravenously injected to mice in the first group, and physiological saline to mice in the second group. The dose was 50 g/kg. Their toxicity manifestation, morality and the change of weight were recorded.② The standard curve of proliferation and metabolism of L929 cells was established. ③ The cytotoxinic test consists of 3 groups: materials group (extracts of the materials), positive control group (culture fluid with 0.64% phenol), and negative control group (RPMI-1640 culture fluid). Each of three was cultured with cell suspension, and then the morphology of the cells was observed, the relative proliferation rate (RGR) was calculated, and the toxicity was classified. ④ In vitrohemolytic test was divided into 3 groups: extracts, sterile distilled water (positive control) and 0.9% physiological saline. In each of three, 0.2 ml anticoagulant diluted fresh rabbit blood was added. The percentage of hemolysis was tested. ⑤ The muscle and implantation test were divided into 4 groups(n=3). The composite biomaterials were implanted into pygal muscleson either side and lateral condyles of femurs. After surgery, the rats of four groups were sacrificed at 12 and 24 weeks respectively.Tissue slice and scanning electronic microscopy were performed. Results General acute toxic test: no mouse died within 3 weeks; no toxicity symptom or adverse effects were shown within 3 days. The weight of materials group increased by 3.57±0.49 g, and the control group by 3.62±0.61 g, showing no statistically significant difference（Ρgt;0.05）.The standard curve of L929 cell perliferation and metabolism showed that their existed a positive correlation between the number of L929 cells and the perliferation. ③ Cytotoxinic test: cytosomes in the positive control group diminished and appeared round, there were pyknotic nucleus, the attached cells agglomerated; the toxicity was level Ⅳ. The morphology of cells in materials groupand negative control group was normal, and the number of them increased; the toxicity was level Ⅰand level 0, respectively. The MTT color experiments showed that positive control group was significantly lower than materials group and negative control group, showing statistically significant difference (Plt;0.01); there was no statistically significant difference between materials group and negative group.④ Hemolytic test: in vitrohemolytic rate of negative control group was0, of positive control group was 100%, and of materials group was 1.66%, which accords with the standard that hemolytic rate should be lower than 5% specified in ISO. ⑤ Implant test:No apparent rejection reaction took place after the composite was implanted; the composite bonded with the bones of the receptors firmly, which had good bonedinduced effect. Conclusion Graded ZrO2-HA composite bioceramic has good biocompatibility and is suitable for orthopedic biomaterials.
目的:研究大豆异黄酮对D半乳糖致衰老大鼠抗氧化能力的影响。方法:用D半乳糖注射Wistar雄性大鼠5个月,建立衰老模型。对致衰老模型组、大豆异黄酮组肝脏、心脏和前列腺丙二醛(MDA)含量、超氧化物歧化酶(SOD)、谷胱甘肽过氧化酶(GSHPx)活性进行测定及比较。结果:低、中、高不同剂量大豆异黄酮灌喂组与模型组大鼠相比,各脏器MDA含量(μmol/L)(心脏:695±093,562±112,435±112比802±111;肝脏:815±085,647±120,515±112比935±135;前列腺:715±092,558±115,423±125比833±124)均有降低,差异有统计学意义(Plt;005),而SOD酶活性(nmol/L)(心脏:4732±308,5518±428,6120±368比3225±370;肝脏:18121±506,19015±706,19720±570比17213±512;前列腺:4156±301,4607±421,5015±335比3374±305)和GSHPx酶活性(nmol/L)(心脏:905±096,1111±245,1313±146比713±151;肝脏:902±105,1150±223,1362±192比698±160;前列腺:435±085,613±102,747±155比312±106)有升高,差异同样具有统计学意义(Plt;005);大豆异黄酮摄入量越高,MDA含量越低,而SOD、GSHPx酶活性越高。结论:摄入适量大豆异黄酮可有效增强大鼠机体抗氧化能力,从而延缓D半乳糖诱发的大鼠衰老。
【摘要】 目的 探讨慢性缺氧对大鼠岩神经节神经元酸敏感离子通道（acid-sensing ion channels，ASICs）亚型3（ASIC3）和亚型2a（ASIC2a）表达的影响。 方法 将12只健康成年SD大鼠随机分为正常组和缺氧组。用免疫组织化学法（PV）观察正常和慢性缺氧大鼠岩神经节神经元ASIC3和ASIC2a的表达。 结果 给予慢性缺氧刺激后，岩神经节ASIC3阳性表达神经元数目增多（Plt;0.05），灰度值降低（Plt;0.05）；而ASIC2a阳性表达神经元数目和灰度值无明显变化（Pgt;0.05）。 结论 慢性缺氧可上调大鼠岩神经节神经元ASIC3的表达，而对ASIC2a的表达无明显影响，提示ASIC3和ASIC2a可能在岩神经节对缺氧的反应中起着不同的作用。【Abstract】 Objective To investigate the effects of chronic hypoxia on expression of acid-sensing ion channels (ASIC) 3 and ASIC2a in neurons of petrosal ganglions of rats. Methods A total of 12 SD rats were randomly assigned to control group and hypoxia group. The expressions of ASIC3 and ASIC2a of the neurons in the petrosal ganglions in the two groups were investigated with the immunohistochemical technique. Results The level of positive ASIC3 expression in the petrosal ganglions was higher in the hypoxia group than that in the control group (Plt;0.05); the difference of positive ASIC2a expression levels between the control group and the hypoxia group was not statistically significant (Pgt;0.05). Conclusion Chronic hypoxia can significantly increase the expression of ASIC3, but not that of ASIC2a, of the neurons in the petrosal ganglions, suggesting their different roles in mediating a cellular response to chronic hypoxia.
ObjectiveTo observe the effects of Rhodiola on the rat retinal tissue morphology and the hypoxia-inducible factor (HIF)-1α at simulated hypoxia at different altitudes. Methods Forty-eight adult female Sprague Dawley rats were randomly divided into the Rhodiola Intervention group (intervention group) and the control group, each group had 24 rats. The intervention group rats were treated with intraperitoneal injection of 10 ml/kg of large plants Rhodiola solution, and the control group rats were injected with same volume of saline. One hour after the injection, six rats were randomly selected from both of the two groups and reared in the plateau environment simulation laboratory modules with the oxygen partial pressure of 17.4, 14.6, 11.3 and 7.4 kPa, which simulated the altitudes of 1500, 3000, 5000 and 8000 meters indoor respectively. Six hours later the rat eyeballs were harvested for paraffin sections and analyzed by hematoxylin and eosin staining, and immunohistochemical staining to observe the expression of HIF-1α and p53. ResultsIn the control group, the rat retinal layers were edema and loose, the retinal thickness increased, the retinal structure was disorganized, the ganglion cells were swollen and degenerated, and some can observe the karyopyknosis, karyolysis and the reduced cells number. As the altitude increased, the pathological changes of retinal became more obvious. In the intervention group, the characteristics of rat retinal morphology were same with the control group, while the degree of morphology changes was lighter than the control group. HIF-1α and p53 expressed mainly in the ganglion cell layer and inner nuclear layer of rat retina in the control group. As altitude increased, the expression of HIF-1α and p53 were increased too, which was positive correlated (r=0.9846, P < 0.05). Compared with the control group, the rat retinal expression of HIF-1α increased, while expression of p53 decreased in the intervention group, and the differences were statistically significant (P < 0.05). ConclusionRhodiola can reduce the retinal tissue pathology damage caused by high altitude hypoxia, and its mechanism may be related to the increasing expression of HIF-1α and reducing expression of p53.
ObjectiveTo systematically review the clinical efficacy and effects on pregnancy outcomes of S-adenosy-L-methionine combined with ursodesoxycholic acid in the treatment of intrahepatic cholestasis of pregnancy. MethodsDatabases such as PubMed, The Cochrane Library, CNKI, VIP, WanFang Data were searched for the studies about the clinical efficacy and effects on pregnancy outcomes of S-adenosy-L-methionine combined with ursodesoxycholic acid in the treatment of intrahepatic cholestasis of pregnancy up to December 31st, 2013. Two reviewers independently screened literature, extracted data and evaluated methodological quality. Then meta-analysis was conducted using RevMan 5.0.24 software. ResultsA total of 11 RCTs involving 776 patients were included. The results of meta-analysis showed that, combined medication reduced blood biochemical indexes inlcuding ALT (MD=3.63, 95%CI 0.63 to 6.64, P=0.02), TB (MD=3.70, 95%CI 1.45 to 5.96, P=0.001), and AST (MD=7.61, 95%CI 2.47 to 12.75, P=0.004). Combined therapy significantly decreased the rates of amniotic fluid contamination (OR=0.29, 95%CI 0.19 to 0.45, P=0.000 01), cesarean section (OR=0.53, 95%CI 0.36 to 0.79, P=0.002), postpartum hemorrhage (OR=0.32, 95%CI 0.12 to 0.90, P=0.03), preterm birth (OR=0.36, 95%CI 0.24 to 0.55, P < 0.000 01), fetal distress (OR=0.33, 95%CI 0.19 to 0.58, P=0.000 1) and neonates asphyxia (OR=0.30, 95%CI 0.19 to 0.47, P < 0.000 01). Combined therapy was also beneficial to improving pruritus symptoms (MD=0.20, 95%CI 0.08 to 0.31, P=0.000 08) and benefiting fetus growth (MD=0.45, 95%CI 0.23 to 0.66, P < 0.000 1). ConclusionThe combination of S-adenosy-L-methionine and ursodesoxycholic acid is superior to ursodesoxycholic acid alone in improving clinical symptoms and pregnant outcomes of patients with intrahepatic cholestasis of pregnancy.
Objective To observe the effects of culture medium of amniotic cells on NO and NOS in retinal tissues of rabbits in vitro in order to provide a protective method for antioxidation in retina transplantation. Methods Thirty adult healthy rabbits (30 right eyes) were divided into 3 groups. Group I: fresh retinal tissue; group II: routine culture medium; group III: culture medium of amniotic cells. The retinal tissues in group II and III were cultured in the corresponding culture medium for 1 week. The content of NO and NOS in retinal tissues in the 3 groups were determined. Results Compared with group I, the content of NO and NOS of group II increased obviously (t=3.821, 3.854; P＜0.001). There was no statistical difference of content of NO and NOS between group I and III (t=1.657, 1.745; P＞0.05). Conclusion Culture medium of amniotic cells may remove free radicals and enhance the ability of antioxidation. (Chin J Ocul Fundus Dis，2004，20：366-368)
ObjectiveTo investigate the expression of keratinocyte growth factor (KGF) and cyclooxygen-ase-2 (COX-2) protein and microvessel density (MVD), and to explore their function and mechanism in the multistep process of gastric cancer. MethodsThe expressions of KGF and COX-2 protein in 64 samples of gastric cancer and 30 cases of normal gastric mucosa tissues were detected by immunohistochemistry. The MVD was detected by staining the endothelial cells in microvessles using anti-CD34 antibody. ResultsThe positive rate of KGF and COX-2 protein expression in gastric cancer were 65.6% (42/64) and 79.7% (51/64), respectively, which was significantly higher than that in normal gastric mucosa tissues 〔(23.3%, 7/30), P=0.046; (13.3%, 4/30), P=0.008〕. The MVD of gastric cancer was 31.8±8.0, which was significantly higher than that of normal gastric mucosa tissues (14.3±6.1), P=0.000. The MVD in gastric cancer with coexpressive KGF and COX-2 protein was 35.9±5.7, which was significant higher than that with non-coexpressive KGF and COX-2 protein (25.7±7.0), P=0.000. Both the expression of KGF and COX-2 protein were related to the invasion of serosa, lymph node metastasis and TNM staging (Plt;0.05, Plt;0.01). The MVD of gastric cancer tissues was related to lymph node metastasis and TNM staging (Plt;0.05), but unrelated to patient’s age, gender, and differentiation of tumor (Pgt;0.05). The co-expression of KGF and COX-2 protein was frequently found in patients with deeper invasion of serosa, lymph node metastasis, and higher TNM staging (Plt;0.05), but which was not associated withpatient’sage, gender, and differentiation of tumor (Pgt;0.05). The expression of KGF protein was positively correlated to the expression of COX-2 protein (r=0.610, P=0.000). There was positive correlation between MVD and the expression of KGF (r=0.675, P=0.000) and COX-2 protein (r=0.657, P=0.000) in gastric cancer, respectively. ConclusionKGF and COX-2 highly expressed by gastric cancer, which may be involved in the invasion and metastasis of gastric cancer by synergisticly promoting the angiogenesis.
OBJECTIVE To study the protective effects of Schwann cell derived neurotrophic factor (SDNF) on motoneurons of spinal anterior horn from spinal root avulsion induced cell death. METHODS Twenty SD rats were made the animal model of C6.7 spinal root avulsion induced motoneuron degeneration, and SDNF was applied at the lesion site of spinal cord once a week. After three weeks, the C6.7 spinal region was dissected out for motoneuron count, morphological analysis and nitric oxide synthase (NOS) enzyme histochemistry. RESULTS 68.6% motoneurons of spinal anterior horn death were occurred after 3 weeks following surgery, the size of survivors was significantly atrophy and NOS positive neurons increased. However, in animals which received SDNF treatment, the death of motoneurons was significantly decreased, the atrophy of surviving motoneurons was prevented, and expression of NOS was inhibited. CONCLUSION SDNF can prevent the death of motoneurons following spinal root avulsion. Nitric oxide may play a role in these injury induced motoneuron death.
Objective To observe the expression of matrix metalloproteinase-9 (MMP-9), its tissue inhibitor of matrix metalloproteinase (TIMP-1), inducible nitric oxide synthase (iNOS) and contents of nitric oxide (NO) in the ocular tissues of Sprague-Dawley (SD) rats with endotoxin induced uveitis（EIU）. Methods Ninety SD rats were randomly divided into experimental (81 rats) and control group (9 rats). The model of EIU was induced in rats in experimental group by injecting with lipoplysaccharide (LPS) 200 μl into the hind feet pads, while the rats in the control group were not injected. Nine rats were executed 0, 6, 12, 18, 24, 48, 72, 96 hours and 7 days, respectively, after injecting with LPS; the NO content and concentration of protein in the aqueous humor in blood plasma, aqueous humor, and uveal tissues were detected. The expressions of MMP-9, TIMP-1 and iNOS in the ocular tissues were detected by immunohistochemistry, and the average absorbance (A) value was evaluated by computer medical image analysis system. Results iNOS, MMP-9 and TIMP-1 expressed in the epithelial cells of iris and ciliary body and exudated inflammatory cells of rats. The concentration of protein in the aqueous humor, the contents of NO in blood plasma, aqueous humor, and uveal tissues, and A value of MMP-9 had obvious relativity with the inflammatory extent, while no positive correlation was found between the inflammatory extent and the A value of iNOS and TIMP-1. Expression of iNOS was found 6 hours after injection, reached the peak after 12 hours, and then dropped gradually. The expression of TIMP-1 could be seen 24 hours after injection, and reached its peak after 72 hours. Conclusion The content of NO and expressions of iNOS, MMP-9 and TIMP-1 changes from the beginning and during the development of EIU, which suggests that NO, iNOS, MMP-9 and TIMP-1 are involved in the pathologic process of EIU. (Chin J Ocul Fundus Dis, 2005, 21: 371-374)