Human platelet lysate supports ex vivo expansion and enhances osteogenic differentiation of human bone marrow-derived mesenchymal stem cells

MSCs (mesenchymal stem cells) with their versatile growth and differentiation potential are ideal candidates for use in regenerative medicine and are currently making their way into clinical trials, which requires the development of xeno-free protocols for their culture. In this study, MSCs were cultured in 10% FCS or 7.5% HPL (human platelet lysate)-supplemented media. We found that both groups of MSCs showed a comparable morphology, phenotype and proliferation. The percentage of cells in the S- and G2-/M-phases, however, was slightly up-regulated (P<0.01) in HPL group. HPL contains PDGF (platelet derived growth factor)-AB and IGF (insulin-like growth factor)-1. In addition, compared with FCS group, MSCs in HPL group showed an increase in osteogenic differentiation and a decrease in adipogenic differentiation. In conclusion, MSCs in HPL-supplemented media maintained similar growing potential and phenotype, while osteogenic potential was enhanced. HPL offers a promising alternative to FCS for MSC expansion for clinical application, especially in bone injury diseases.     

Integrin alphav-mediated inactivation of p53 controls a MEK1-dependent melanoma cell survival pathway in three-dimensional collagen

Integrin alphav is required for melanoma cell survival and tumor growth in various models. To elucidate integrin alphav-mediated melanoma cell survival mechanisms, we used a three-dimensional (3D) collagen gel model mimicking the pathophysiological microenvironment of malignant melanoma in the dermis. We found that integrin alphav inactivated p53 and that suppression of p53 activity by dominant negative p53 or p53-small interfering RNA obviated the need for integrin alphav for melanoma cell survival in 3D-collagen and for tumor growth in vivo. This indicates that integrin alphav-mediated inactivation of p53 functionally controls melanoma cell survival. Furthermore, we found that melanoma cell integrin alphav was required for MAPK kinase (MEK) 1 and extracellular signal-regulated kinase (ERK)1/2 activity in 3D-collagen, whereas inhibition of MEK1 activity induced apoptosis. Surprisingly, MEK1 and ERK1/2 activities were restored in integrin alphav-negative melanoma cells by suppression of p53, whereas concomitant block of MEK1 induced apoptosis. This suggests that integrin alphav controls melanoma cell survival in 3D-collagen through a pathway involving p53 regulation of MEK1 signaling.     

假苍耳的生活史进程中几种生理生化指标的变化

本文试图从生理生化的角度对假苍耳(Iva xanthifolia)生活史进程中可溶性糖类、赤霉素、单宁以及黄酮的变化进行探讨。通过对假苍耳在生长发育期间几种生理生化指标的测定, 结果表明, 在假苍耳生活史进程的不同阶段, 其体内各种代谢产物的含量基本都在种子或芽阶段具有最高含量。此外, 不同发育阶段可溶性还原糖含量的变化规律相似: 芽>花序>苗>成株>种子。除在花序和苗阶段没有测到海藻糖, 其他各阶段海藻糖的含量变化如下: 芽>成株>种子。另外, 只有在种子阶段检测到棉子糖, 其含量为15.43 mg.g-1。赤霉素含量的变化规律如下: 种子＞芽≈苗≈花序＞成株。单宁含量的变化趋势: 种子＞成株＞苗＞芽＞花序。黄酮含量的变化趋势: 种子＞芽＞成株≈花序＞苗。值得注意的是, 当单宁/黄酮的比值接近1时, 植物体内需要的单宁和黄酮的含量则相对较低; 相反, 当单宁/黄酮的比值接近0时, 植物体内需要的单宁和黄酮的含量则较高。     

Neuroprotection of Pramipexole in UPS Impairment Induced Animal Model of Parkinson’s Disease

Pramipexole (PPX), a dopamine (DA) receptor D3 preferring agonist, has been used as monotherapy or adjunct therapy to treat Parkinson’s disease (PD) for many years. Several in vitro and in vivo studies in neurotoxin-induced DA neuron injury models have reported that PPX may possess neuroprotective properties. The present study is to evaluate the neuroprotection of PPX in a sustained DA neuron degeneration model of PD induced by ubiquitin–proteasome system (UPS) impairment. Adult C57BL/6 mice were treated with PPX (low dose 0.1 mg/kg or high dose 0.5 mg/kg, i.p, twice a day) started 7 days before, and continued after microinjection of proteasome inhibitor lactacystin in the medial forebrain bundle for a total 4 weeks. Animal behavior observation, and pathological and biochemical assays were conducted to determine the neuroprotective effects of PPX. We report here that PPX treatment significantly improves rotarod performance, attenuates DA neuron loss and striatal DA reduction, and alleviates proteasomal inhibition and microglial activation in the substantia nigra of lactacystin-lesioned mice. PPX can increase the levels of brain-derived neurotrophic factor and glial cell line-derived neurotrophic factor and induce an activation of autophagy. Furthermore, pretreatment with D3 receptor antagonist U99194 can significantly block the PPX-mediated neuroprotection. These results suggest that multiple molecular pathways may be attributed to the neuroprotective effects of PPX in the UPS impairment model of PD.     

鼠肝中金属硫蛋白的纯化

旨在探索鼠肝中金属硫蛋白(MT)提取工艺并加以改进。按照经典方法工艺提取MT,用中空纤维柱超滤方法加以改进,依据MT自身特点进行分离和鉴定。结果显示,粗品符合MT特点:分子量在6.5kD左右;无280nm特征吸收峰,加酸后紫外吸收(200-300nm)肩峰消失;通过离子交换可以实现MT1、MT2的分离;通过原子吸收测定,含有锌、铜、镉3种金属,锌含量最高,具有重要的病理生理意义。因此,与其它方法比较,改进后方法更适宜实验室制备。     

大薯ISSR-PCR反应体系的优化

以大薯为材料,通过试验优化了ISSR-PCR反应体系中的主要成分用量及浓度、退火温度对大薯ISSR扩增结果的影响。试验表明:在20μL的反应体系中,Mg2+的浓度为1.875 mmol/L,dNTPs浓度为0.500 mmol/L,引物的浓度为1.0μmol/L,TaqDNA聚合酶的用量为1U,模板DNA的用量为60 ng,在48℃的退火温度下45个循环,能得到清晰、多态性高的ISSR带谱。     

Synthesis of 3-O-(beta-D-xylopyranosyl-(1-->2)-beta-D-glucopyranosyl)-3'-O-(beta-D-glucopyranosyl)tamarixetin, the putative structure of aescuflavoside A from the seeds of Aesculus chinensis

3-O-(beta-D-xylopyranosyl-(1-->2)-beta-D-glucopyranosyl)-3'-O-(beta-D-glucopyranosyl)tamarixetin, the putative flavonal glycoside named aescuflavoside A, isolated from the seeds of Aesculus chinensis, is synthesized via regioselective glycosylation of 7-O-benzyltamarixetin with glycosyl bromides under phase-transfer-catalyzed conditions.     

RDMAS: a web server for RNA deleterious mutation analysis

Background  

The diverse functions of ncRNAs critically depend on their structures. Mutations in ncRNAs disrupting the structures of functional sites are expected to be deleterious. RNA deleterious mutations have attracted wide attentions because some of them in cells result in serious disease, and some others in microbes influence their fitness.     

酸性土壤中高效半纤维素降解菌的筛选与鉴定

【目的】筛选能够适应南方酸性土壤的高效半纤维素降解菌株,并进行鉴定,确定菌株的安全性。【方法】采用半纤维素平板水解圈法和胞外酶测定法进行菌株筛选,通过拮抗实验构建复合微生物菌系。利用培养特征、形态学、生理生化特征和分子生物学方法进行菌株鉴定。【结果】筛选出效果稳定,互不拮抗的高效半纤维素降解放线菌4株(NA9、NA10、NA12和NA13),半纤维素酶活分别为:217.6、229.8、221.1和211.8 U/mL。真菌2株NF1和NF7,半纤维素酶活为217.7和244.2 U/mL。复合微生物菌系半纤维素酶活可达299.0 U/mL。经鉴定菌株NA9、NA10、NA12和NA13为链霉菌中的哥斯达黎加链霉菌(Streptomyces costaricanus)。菌株NF1为亮白曲霉(Aspergillus candidus),菌株NF7为黄蓝状菌(Tarlaromyces flavus)。