Guided bone regeneration with beta-tricalcium phosphate and poly L-lactide-co-glycolide-co-epsilon-caprolactone membrane in partial defects of canine humerus

This study was performed to evaluate the effect of beta-tricalcium phosphate and poly L-lactide-co-glycolide-co-epsilon-caprolactone (TCP/PLGC) membrane in the repair of partial bone defects in canine proximal humerus. Three adult mixed-breed dogs were used during the experimental period. The length of the defect was quarter of the full length of humerus, and width of the defect was quarter of middle diameter of the lateral aspect of humerus. The humeri of each dog were divided into treatment (TCP/PLGC) and control groups. The defect was covered with TCP/PLGC membrane in treatment group. To evaluate regeneration of the bone, computerized tomography (CT) and histopathologic examination were performed. The radiopaque lines were appeared at the original defect sites in TCP/PLGC group but below the original site in control at 4th week. Radiopacity and thickness of the defect sites, and radiopaque lines were more increased at 8th week than those of 4th week. Histopathologic findings revealed fibrous connective tissue migration into the defect and the migration inhibited the structure of new cortex to be placed in the original level in control whereas new cortex growth was found in the level of original line in TCP/PLGC group. However, the new cortical bone in the TCP/PLGC group was thinner and less organized than the adjacent intact cortex, and the amount of new cancellous bones were also scanty. The result suggested that TCP/PLGC membrane is a good guided bone regeneration material to restore the original morphology of humerus in partial defect.     

May antioxidants status depletion by Tetradifon induce secondary genotoxicity in female Wistar rats via oxidative stress?

Tetradifon is a potent organochlorine acaricide with an estrogen-like structure. The wide spread use of this chemical is likely to pollute the environment. Only few studies have been reported for the evaluation of its short- and long-term toxic effects including genotoxicity and carcinogenicity and there have been conflicting results in in vitro and in vivo test systems. In this work, we have evaluated Tetradifon for its ability to induce genotoxic damages in female Wistar rats. A single cumulative dose of 2430 mg/kg BW was administrated orally for 12 female rats of 190 g BW during 12 weeks. Twelve additional rats, no treated, have served as control. Animals were sacrificed after 6 and 12 weeks of treatment. Genetic toxicity studies were conducted in rats bone marrow, by chromosomal aberrations (CAs) and sister-chromatid exchanges (SCEs) assays. The oxidative stress status of treated animals has been also evaluated by assessment of lipid peroxidation by measuring thiobarbituric acid reactive substances (TBARS). Some serum parameters: vitamins (A and E), triglyceride (TG) and total antioxidants status (TAS) were determined. Our results showed a significant increase in tissue TBARS concentrations in the two treated groups suggesting that Tetradifon induce an oxidative stress. Elsewhere, rats treated with Tetradifon exhibited a statistical decrease in serum level of vitamin E and a significant depletion of serum total antioxidant status. Whereas, in comparison to control rats, treated animal cells did not show a significant increase in either the frequency of SCEs or CAs. These results indicate that Tetradifon did not present direct genotoxic effect in female Wistar rats. But we suggest that its inducting of an oxidative stress may lead to indirect mutagenecity that should be evaluated by other series of in vivo genotoxicity assays as micronucleus test or comet assay.     

牛GDF9和BMP15基因遗传变异与双胎性状的关系研究

以生长分化因子9(Growth differentiation factor 9,GDF9)基因和骨形态发生蛋白15(Bone morphogenet-ic protein 15,BMP15)基因作为牛双胎性状的候选基因,研究了它们在鲁西牛、秦川牛、南阳牛和中国荷斯坦牛4个品种中的遗传变异,并在鲁西牛群体中研究了其多态位点与双胎性状的关系。结果表明:在鲁西牛中GDF9基因的3′UTR发现缺失突变,而其它3个品种中没有发现该突变。对鲁西牛群体中该多态位点与单、双胎性状之间进行卡方显著性检验表明,单胎牛群体与双胎牛群体基因型分布有极显著的差异(P=0.006),双胎牛群体的B等位基因频率明显大于单胎牛群体。通过生物信息学分析表明,突变体mRNA的二级结构与野生型相比总自由能值差异不大,但突变体mRNA翻译起始位点的二级结构稳定性明显大于野生型。在鲁西牛、南阳牛和秦川牛的BMP15基因中发现编码区第759~762位有GAAA 4个碱基存在缺失突变,但没有检测到突变纯合个体,中国荷斯坦牛中没有检测到该突变。卡方显著性检验表明单胎牛群体和双胎牛群体在该位点基因型组成差异不显著(P=0.947)。     

食蟹猴骨髓采集穿刺方法及其骨髓象测定

为探索采集食蟹猴骨髓的操作方法及其正常骨髓象指标,试验采集了60只食蟹猴的髂骨骨髓,并测定其骨髓象、测得的骨髓象及采集过程中的注意事项.结果表明,可以顺利的采出骨髓,雌雄之间晚幼红细胞、粒红比有显著性差异(P＜0.05),其余无显著差异.     

Isolation and identification of human fetal bone-derived mesenchymal stem cells and their differentiation to hepatocyte like cells

AIM: To separate and identify the mesenchymal stem cells (MSCs) from human fetal bone and to study their differentiation to hepatocyte like cells under the action of chemical induction.METHODS: The MSCs from human fetal bone were isolated and purified according to the different growth characteristic of attaching to the wall of cell culture flask.The cell cycle and surface markers of MSCs were identified using flow cytometry.The MSCs were pre-induced by adding DMSO,β-Me and 5-aza for 24 h,then adding the inductive medium of H-DMEM and rh-HGF to induce their differentiation to hepatocyte like cells (HLCs).HLCs were identified by the typical morphological change and the expression of special protein with the method of immunocytochemistry.RESULTS: The MSCs derived from human fetal bone expressed adhesion molecules CD29+,CD44+,but not antigens of hematopoietic CD34,CD45,and not antigens related to GVHD,such as HLA-DR,CD80 and CD86.Exposure of these cells to above-mentioned inductive agents resulted in obvious morphological change and an increase in expression of AFP and ALB.CONCLUSION: The results suggest the existence of plentiful MSCs in human fetal bone.MSCs derived from human fetal bone can easily differentiate to HLCs,and they have a lower immunogenic nature,which may provide the ideal source for tissue engineering (bioartificial liver) for cellular therapeutics.     

Normal haematopoiesis,cellular components and stainable iron content in the bone marrow of camels (Camelus dromedarius)

Bone marrow samples were collected from the ribs of 20 healthy adult male Iranian camels (Camelus dromedarius). The bone marrow smears were stained using Wright's stain. Blood samples were collected at the same time for routine haematological examination. The development and morphology of the blood cells in the bone marrow of the camels were similar to those of other domestic species. The mean myeloid/erythroid ratio was 1.21, the mean erythroid percentage was 42.7% and the mean myeloid percentage was 52.0%. The mean percentage of other cells was 5.3%. The mean percentage of eosinophils in the myeloid series was higher than in other domestic species. Iron stores, estimated from Perl's stain, ranged from scant (1+) to moderate (3+) but most samples had 2+ iron content. All peripheral blood results were within reference ranges.     

绵羊骨代谢相关基因VDR密码子偏好性分析

【目的】明确绵羊维生素D受体(vitamin D receptor,VDR)基因密码子的使用特征,探究不同物种VDR基因进化规律,为骨代谢等相关疾病动物建模提供参考依据。【方法】利用CodonW、EMBOSS、Usage Codon和DNAMAN等软件对VDR基因CDS区的同义密码子相对使用度(RSCU)、密码子碱基含量、有效密码子数(ENc)及密码子偏好指数(CBI)等参数进行统计分析,并在多物种间进行比较,同时利用ENc-Plot关联分析和中性绘图等方法分析VDR基因密码子偏好性使用的主要影响因素,并基于多物种同义密码子相对使用度数据与系统进化聚类分析探究密码子使用模式与生物进化的内在联系。【结果】绵羊VDR基因中有23个偏好使用以G/C结尾的密码子;大猩猩、鸸鹋、褐家鼠等其他14个物种在密码子使用上表现出了与之较为相似的特征,而在半胱氨酸、甘氨酸、组氨酸、丝氨酸、脯氨酸等个别氨基酸编码时,鸸鹋、褐家鼠和麻雀等部分物种显现出独有特点;ENc-Plot绘图和中性绘图分析显示,选择压力对VDR基因密码子偏好性的影响强于突变压力。【结论】以G/C结尾的密码子为VDR基因优势密码子,自然选择是导致其偏好性的主要力量,密码子偏好使用存在物种特异性,并与物种亲缘关系进化存在关联性。     

麦秸半焦低温催化气化实验研究

以麦秸半焦为研究对象,比较麦秸半焦和酸洗麦秸半焦700℃下水蒸气气化实验,研究了麦秸灰分中碱金属含量、催化剂种类(K基、Ca基和Fe基)对气化过程的影响。实验结果表明,麦秸灰分中较高含量的碱金属K能够有效促进半焦气化的进行,使得在非催化条件下半焦的碳转化率达到63.4%,相比酸洗麦秸半焦提高了2.6倍。低温下碱金属K基催化剂对半焦气化的促进作用显著,在700℃和K/Al催化剂条件下H2产率和碳转化率分别达到118.60 mol/kg和98.5%,相较于Ca/Al、Fe/Al催化剂而言,H2产率分别提高了10.7%和20.5%,碳转化率提高了12.4%和31.0%;而Ca/Al及Fe/Al催化剂有利于水气变换反应的进行,可提高气化产物中H2体积分数,降低CO体积分数。综合考虑碳转化率、气体产率及气体组分含量等因素,在催化效果较好的K基催化剂中添加适量的Ca基催化剂有利于调节产气成分,从而进一步提纯获得富氢气体。     

Differentiation of canine bone marrow stromal cells into voltage- and glutamate-responsive neuron-like cells by basic fibroblast growth factor

We investigated the in vitro differentiation of canine bone marrow stromal cells (BMSCs) into voltage- and glutamate-responsive neuron-like cells. BMSCs were obtained from the bone marrow of healthy beagle dogs. Canine BMSCs were incubated with the basal medium for neurons containing recombinant human basic fibroblast growth factor (bFGF; 100 ng/ml). The viability of the bFGF-treated cells was assessed by a trypan blue exclusion assay, and the morphology was monitored. Real-time RT-PCR was performed to evaluate mRNA expression of neuronal, neural stem cell and glial markers. Western blotting and immunocytochemical analysis for the neuronal markers were performed to evaluate the protein expression and localization. The Ca²⁺ mobilization of the cells was evaluated using the Ca²⁺ indicator Fluo3 to monitor Ca²⁺ influx. To investigate the mechanism of bFGF-induced neuronal differentiation, the fibroblast growth factor receptor inhibitor, the phosphoinositide 3-kinase inhibitor or the Akt inhibitor was tested. The bFGF treatment resulted in the maintenance of the viability of canine BMSCs for 10 days, in the expression of neuronal marker mRNAs and proteins and in the manifestation of neuron-like morphology. Furthermore, in the bFGF-treated BMSCs, a high concentration of KCl and L-glutamate induced an increase in intracellular Ca²⁺ levels. Each inhibitor significantly attenuated the bFGF-induced increase in neuronal marker mRNA expression. These results suggest that bFGF contributes to the differentiation of canine BMSCs into voltage- and glutamate-responsive neuron-like cells and may lead to the development of new cell-based treatments for neuronal diseases.     

Plasma fluctuation in estradiol-17β and bone resorption markers around parturition in dairy cows

Blood samples were obtained sequentially from 10 dairy cows around the time of parturition to assess plasma fluctuations in estradiol-17β (E₂) levels in association with those of several bone resorption markers. Plasma E₂ concentration increased sharply a few days prepartum and decreased quickly after parturition. In terms of bone resorption markers, the plasma level of tartrate-resistant acid phosphatase isoform 5b (TRAP5b) rose significantly, commencing 1 week prepartum, and was maintained at this level to a few days postpartum. The plasma concentration of carboxyterminal collagen cross-links of type-I collagen (CTx) increased significantly after parturition. These observations suggest that osteoclast-mediated bone resorption was activated after parturition when plasma E₂ concentrations decreased.