The effect of matrix stiffness on the chondrogenic differentiation of mesenchymal stem cells

Journal of Molecular Histology - Articular cartilage is one of the most important weight-bearing components in human body, thus the chondrogenesis of stem cells is reactive to many intracellular...     

Human overexploitation and extinction risk correlates of Chinese snakes

China is one of the countries with the richest snake biodiversity in the world. However, about one‐third of all 236 species are now considered threatened, partially due to the intense human overexploitation. Despite that, to date, no study has explicitly investigated the patterns and processes of extinction and threats of Chinese snakes, or between human exploited and unexploited snake subgroups. We addressed the following three questions: 1) which snake families proportionally include more human exploited species than expected by chance? 2) Which species traits and extrinsic factors are correlated with their extinction risk? 3) Are there differences between human exploited and unexploited species in terms of patterns and processes of extinction? We found that the family Elapidae contained a significantly higher number of exploited species. Considering eight species traits and four extrinsic factors, we performed phylogenetic correlation tests, finding that small geographic range size, large body length, oviparous reproduction, diurnal activity and high human exploitation were important in determining the extinction risk of all Chinese snakes. Moreover, human exploited snakes had a higher percentage of threatened species and large‐bodied species than unexploited snakes. Extinction risk of human exploited species was related to body length, reproduction mode and activity period, whereas that of human unexploited species were associated with geographic range size, microhabitat and annual temperature. Overall, we highlight the phylogenetic non‐random exploitation of snakes, and different factors underlying species response to human overexploitation. We suggest that conservation priority should be given to exploitation‐prone families and species with extinction‐prone traits, as identified in this study. Moreover, human exploited and unexploited species should be managed considering different strategies since their extinction risk was associated with different ecological traits. Conservation actions should also focus on preventing human threats, such as human overexploitation and habitat loss, for the effective preservation of Chinese snakes.     

青海祁连棱果沙棘自然杂交带三种沙棘 光合特性日变化分析

杂交带是研究物种形成及进化的天然实验室。为探究棱果沙棘自然杂交带三种沙棘的生理生态适应性,该研究在光照充足的7月中旬分别测定了该区域三种沙棘雌、雄株的光合特性及相关环境因子的日变化。结果表明:(1)同种沙棘雌、雄株间光合日变化规律基本一致,不同沙棘光合特性的日变化规律存在较大差异。中国沙棘(Hippophae rhamnoides ssp. sinensis)净光合速率(Pn)日变化呈明显的双峰曲线,峰值出现在光合有效辐射(PAR)和空气温度(Ta)相对较高的10:00和14:00,最高达到(19.53±5.35)μmol·m~(-2)·s~(-1);棱果沙棘(Hippophae goniocarpa)和肋果沙棘(H. neurocarpa ssp. neurocarpa) Pn日变化均呈近双峰曲线,在PAR和Ta相对较低的8:00和16:00有较高的值,最高值分别为(13.43±3.43)和(15.27±2.43)μmol·m~(-2)·s~(-1)。(2)三种沙棘水分利用效率(WUE)与Pn的日变化规律一致,但中国沙棘具有最高的WUE,达到(6.72±0.95)μmol·mmol~(-1),棱果沙棘和肋果沙棘的WUE日变化最高分别为(4.03±1.08)和(4.93±0.86)μmol·mmol~(-1)。(3)三种沙棘蒸腾速率(Tr)、气孔导度(Gs)、胞间二氧化碳浓度(Ci)和气孔限制值(Ls)的日变化规律相似,其中Gs均在10:00后一致下降,在12:00左右均出现不同程度的气孔关闭现象,这是其发生光合午休的主要原因之一。杂交带三种沙棘光合特性日变化差异主要表现在中国沙棘和其他两种沙棘之间,而杂交种棱果沙棘与亲本种肋果沙棘的变化规律基本一致,其Pn与WUE可能受到了Ta、PAR和空气相对湿度(RH)等环境因子的影响。     

Salvianolic acid inhibits the effects of high glucose on vascular endothelial dysfunction by modulating the Sirt1‐eNOS pathway

Salvianolic acid (SA) is known for improving blood circulation, scavenging hydroxyl radicals, and preventing platelet aggregation. The research explored whether SA can protect against cardiovascular disease induced by high glucose conditions. Our results indicate that SA significantly increases cells viability and nitric oxide levels while decreasing reactive oxygen species generation. SA upregulated the expression levels of Bcl‐2 and decreased the levels of Bax, cleaved caspase‐3, and cleaved caspase‐9. Furthermore, the expression levels of Sirtuin 1 (Sirt1) and p‐endothelial nitric oxide synthase (eNOS) were markedly increased in response to SA treatment. Moreover, exposure of human umbilical vein endothelial cells to Ex527 resulted in reducing expression of p‐eNOS. However, the beneficial effects of SA were abolished partially when Ex527 was added. These findings suggest that SA can be used as a potential therapeutic to protect against high glucose‐induced endothelial injury by modulating Sirt1‐eNOS pathway.     

The guanidine hydrochloride-induced denaturation of CP43 and CP47 studied by terahertz time-domain spectroscopy

Terahertz time-domain spectroscopy (THz-TDS) is a new technique in studying the conformational state of a molecule in recent years. In this work, we reported the first use of THz-TDS to examine the denaturation of two photosynthesis membrane proteins: CP43 and CP47. THz-TDS was proven to be useful in discriminating the different conformational states of given proteins with similar structure and in monitoring the denaturation process of proteins. Upon treatment with guanidine hydrochloride (GuHCl), a 1.8 THz peak appeared for CP47 and free chlorophyll a (Chl a). This peak was deemed to originate from the interaction between Chl a and GuHCl molecules. The Chl a molecules in CP47 interacted with GuHCl more easily than those in CP43. Supported by the National Natural Science Foundation of China (Grant No. 39890390)     

Heterologous expression and characterization of recombinant glycerol dehydratase from Klebsiella pneumoniae in Escherichia coli

Glycerol dehydratase (EC 4.2.1.30), as one of the key enzymes in converting glycerol to the valuable intermediate 1,3-propanediol, is important for biochemical industry. The dhaB genes encoding coenzyme B(12)-dependent glycerol dehydratase in Klebsiella pneumoniae were cloned and expressed in Escherichia coli. An effective co-expression system of multiple subunits protein was constructed. Heterologous expression vectors were constructed using the splicing by overlap extension-PCR technique to co-express the three subunits of the glycerol dehydratase. After induction by isopropyl-beta-D-thiogalactopyranoside, SDS-PAGE analysis revealed that: (i) only the alpha subunit of glycerol dehydratase was expressed in direct expression system, (ii) the three subunits of glycerol dehydratase with predicted molecular massess of 64 (agr;), 22 (beta), and 16 kDa (gamma) were expressed simultaneously in co-expression system, and (iii) the fusion expression system expressed the fusion protein of 99 kDa. Enzyme assay showed that the activities of three heterologous expression products were 27.4, 2.3, and 0.2 U/mg. The highest enzyme activity was almost 17 times of that in K. pneumoniae. The recombinant enzyme was purified and biochemically characterized. The apparent Km values of the enzyme for coenzyme B(12) and 1, 2-propanediol were 8.5 nM and 1.2 mM, respectively. The enzyme showed maximum activity at pH 8.5 and 37 degrees C.     

In vitro and in vivo effects of rat kidney vascular endothelial cells on osteogenesis of rat bone marrow mesenchymal stem cells growing on polylactide-glycoli acid (PLGA) scaffolds

It is well established that vascularization is critical for osteogenesis. However, adequate vascularization also remains one of the major challenges in tissue engineering of bone. This problem is further accentuated in regeneration of large volume of tissue. Although a complex process, vascularization involves reciprocal regulation and functional interaction between endothelial and osteoblast-like cells during osteogenesis. This prompted us to investigate the possibility of producing bone tissue both in vitro and ectopically in vivo using vascular endothelial cells because we hypothesized that the direct contact or interaction between vascular endothelial cells and bone marrow mesenchymal stem cells are of benefit to osteogenesis in vitro and in vivo. For that purpose we co-cultured rat bone marrow mesenchymal stem cells (MSC) and kidney vascular endothelial cells (VEC) with polylactide-glycolic acid scaffolds. In vitro experiments using alkaline phosphatase and osteocalcin assays demonstrated the proliferation and differentiation of MSC into osteoblast-like cells, especially the direct contact between VEC and MSC. In addition, histochemical analysis with CD31 and von-Willebrand factor staining showed that VEC retained their endothelial characteristics. In vivo implantation of MSC and VEC co-cultures into rat's muscle resulted in pre-vascular network-like structure established by the VEC in the PLGA. These structures developed into vascularized tissue, and increased the amount and size of the new bone compared to the control group (p < 0.05). These results suggest that the vascular endothelial cells could efficiently stimulate the in vitro proliferation and differentiation of osteoblast-like cells and promote osteogenesis in vivo by the direct contact or interaction with the MSC. This technique for optimal regeneration of bone should be further investigated.     

Capillary electrophoresis coupled with electrochemiluminescence for determination of atomoxetine hydrochloride and the study on its interactions with three proteins

A simple, rapid and sensitive method for the determination of atomoxetine hydrochloride (AH) by capillary electrophoresis with electrochemiluminescence detection (CE‐ECL) using tris(2,2′‐bipyridyl) ruthenium (II) was developed. Under optimized conditions, the determinations of AH in capsules and rat plasmas and the study on its interactions with three plasma proteins, including bovine serum albumin, cytochrome c and myoglobin were performed successfully. Relative to some previous studies, in this paper the methodologies for the determination of AH in aqueous solution and spiked plasma systems were established, respectively. By comparing the difference between the two work curves of two systems, the matrix effect in plasma samples on the determination of AH by the CE‐ECL method was discussed in detail. The results indicated that the effect of the matrix in plasma samples should not be ignored even if no obvious interference was found in the electropherograms and the establishment of method validation in complex samples by the CE‐ECL method was necessary. Copyright © 2014 John Wiley & Sons, Ltd.