Gas7在大鼠关节软骨中的表达及意义

目的探讨生长休止蛋白7（Gas7）在成年大鼠关节软骨中的表达及意义。方法雄性SD大鼠（3个月、12个月）各12只。取膝关节和股骨上端关节部分,经脱钙石蜡包埋并切片,采用HE染色和免疫组化SABC法检测。检测各关节软骨组织中Gas7的表达及其定位。结果关节软骨切线层与移行层Gas7的表达呈现强阳性,而在辐射层及软骨钙化层则弱阳性或阴性。结论Gas7在成年大鼠关节软骨各层细胞的表达不同,可能与各层软骨细胞的生长发育及代谢状况有关     

Lavage with Allicin in Combination with Vancomycin Inhibits Biofilm Formation by Staphylococcus epidermidis in a Rabbit Model of Prosthetic Joint Infection

Background and Aim

The present anti-infection strategy for prosthetic joint infections (PJI) includes the use of antibiotics and surgical treatments, but the bacterial eradication rates are still low. One of the major challenges is the formation of biofilm causing poor bacterial eradication. Recently it has been reported that allicin (diallyl thiosulphinate), an antibacterial principle of garlic, can inhibit bacteria adherence and prevent biofilm formation in vitro. However, whether allicin could inhibit biofilm formation in vivo is unknown. The aim of this study was to investigate the effects of allicin on biofilm formation, and whether allicin could potentiate the bactericidal effect of vancomycin in a rabbit PJI model.

Methods

A sterile stainless-steel screw with a sterile ultra-high molecular weight polyethylene washer was inserted into the lateral femoral condyle of the right hind knee joint of rabbit, and 1 mL inoculum containing 10⁴ colony-forming units of Staphylococcus epidermidis was inoculated into the knee joint (n = 32). Fourteen days later, rabbits randomly received one of the following 4 treatments using continuous lavages: normal saline, vancomycin (20 mcg/mL), allicin (4 mg/L), or allicin (4 mg/L) plus vancomycin (20 mcg/mL). Three days later, the washer surface biofilm formation was examined by scanning electron microscopy (SEM). The bacterial counts within the biofilm of implanted screws were determined by bacterial culture.

Results

The lowest number of viable bacterial counts of Staphylococcus epidermidis recovered from the biofilm was in the rabbits treated with allicin plus vancomycin (P<0.01 vs. all other groups). The biofilm formation was significantly reduced or undetectable by SEM in rabbits receiving allicin or allicin plus vancomycin.

Conclusion

Intra-articular allicincan inhibit biofilm formation and enhance the bactericidal effect of vancomycin on implant surface in vivo. Allicin in combination with vancomycin may be a useful anti-infection strategy for the treatment of PJI.     

A system for the transformation and regeneration of the recretohalophyte Limonium bicolor

Limonium bicolor, a typical recretohalophyte, has a specialized salt-secreting structure in the epidermis called the salt gland and plays a significant role in improving saline land. Understanding the molecular mechanisms of salt secretion and salt gland development requires an efficient L. bicolor transformation system, which is described in this report. Leaf explants were incubated with Agrobacterium tumefaciens strain EHA105 harboring the plasmid pTCK303 containing the β-glucuronidase gene (GUS) as the transgene reporter and the hygromycin B resistance gene as a selectable marker. Up to 96.9% of leaves were induced to regenerate shoots on an Murashige and Skoog (MS) medium supplemented with 4.4 μM 6-benzyladenine and 1.1 μM α-naphthaleneacetic acid; roots were induced on the MS medium containing 2.5 μM indole-3-butyric acid. This tissue culture system was suitable for Agrobacterium-mediated transformation of L. bicolor. Pre-cultivated explants (2 d old) were incubated with Agrobacterium (0.6–0.7 at OD₆₀₀) in a shaking culture for 20 min; the explants and bacterium were co-cultivated for 4 d in the dark before the explants were transferred to a selection medium containing 8 mg/L hygromycin B and 600 mg/L piperacillin sodium (added to prevent continued Agrobacterium growth). Histochemical assays and PCR to detect the GUS gene showed that transformation frequency was 4.43%. Quantitative PCR and Northern blotting further verified the integration and presence of the GUS gene in L. bicolor. This is the first report of an Agrobacterium-based transformation system for L. bicolor. The system will facilitate a research on the identity and function of genes involved in salt gland development and salt secretion.     

应用自组织映射方法的北京市2005-2013年土地利用时空演变分析

时间序列土地利用时空演变规律分析是当前的研究热点之一,通过应用自组织映射神经网络方法进行多时间序列土地利用变化时空一体化表达与演变规律分析,探索区域土地利用变化模式。基于北京市2005、2007、2009、2011、2013年5期土地利用遥感分类数据,构建自组织映射神经网络并利用其聚类和降维可视化功能对5个年份的土地利用数据同时进行训练输出,发现建设用地、耕地、林地、牧草地、园地的聚集模式,并通过对输出神经元进行二次聚类以及土地利用变化轨迹分析,获得北京市郊区5个监测时相土地利用变化的时空演变特征。结果揭示出北京市郊区2005-2013年土地利用变化具有明显的耕地型向建设用地型发展的平原区演变特征,以及向林地型发展的山区演变特征,且各区的发展具有时间上的顺序性;总体上形成6类土地利用演变轨迹。     

Intermittent hydrostatic pressure maintains and enhances the chondrogenic differentiation of cartilage progenitor cells cultivated in alginate beads

The objective of this study was to explore the effects of intermittent hydrostatic pressure (IHP) on the chondrogenic differentiation of cartilage progenitor cells (CPCs) cultivated in alginate beads. CPCs were isolated from the knee joint cartilage of rabbits, and infrapatellar fat pad‐derived stem cells (FPSCs) and chondrocytes (CCs) were included as the control cell types. Cells embedded in alginate beads were treated with IHP at 5 Mpa and 0.5 Hz for 4 h/day for 1, 2, or 4 weeks. The cells' migratory and proliferative capacities were evaluated using the scratch and Live/Dead assays, respectively. Hematoxylin and eosin staining, safranin O staining, and immunohistochemical staining were performed to determine the effects of IHP on the synthesis of extracellular matrix (ECM) proteins. Real‐time polymerase chain reaction analysis was performed to measure the expression of genes related to chondrogenesis. The scratch and Live/Dead assays revealed that IHP significantly promoted the migration and proliferation of FPSCs and CPCs to different extents. The staining experiments showed greater production of cartilage ECM components (glycosaminoglycans and collagen II) by cells exposed to IHP, and the gene expression analysis demonstrated that IHP stimulated the expression of chondrocyte‐related genes. Importantly, these effects of IHP were more prominent in CPCs than in FPSCs and CCs. Considering all of our experimental results combined, we conclude that CPCs demonstrated a stronger chondrogenic differentiation capacity than the FPSCs and CCs under stimulation with IHP. Thus, the use of CPCs, combined with mechanical stimulation, may represent a valuable strategy for cartilage tissue engineering.     

Voltage-gated potassium channel Kv1.3 regulates GLUT4 trafficking to the plasma membrane via a Ca2+-dependent mechanism

Kv1.3 is a voltage-gated K⁺ channel expressed in insulin-sensitive tissues. We previously showed that gene inactivation or pharmacological inhibition of Kv1.3 channel activity increased peripheral insulin sensitivity independently of body weight by augmenting the amount of GLUT4 at the plasma membrane. In the present study, we further examined the effect Kv1.3 on GLUT4 trafficking and tested whether it occurred via an insulin-dependent pathway. We found that Kv1.3 inhibition by margatoxin (MgTX) stimulated glucose uptake in adipose tissue and skeletal muscle and that the effect of MgTX on glucose transport was additive to that of insulin. Furthermore, whereas the increase in uptake was wortmannin insensitive, it was completely inhibited by dantrolene, a blocker of Ca²⁺ release from intracellular Ca²⁺ stores. In white adipocytes in primary culture, channel inhibition by Psora-4 increased GLUT4 translocation to the plasma membrane. In these cells, GLUT4 protein translocation was unaffected by the addition of wortmannin but was significantly inhibited by dantrolene. Channel inhibition depolarized the membrane voltage and led to sustained, dantrolene-sensitive oscillations in intracellular Ca²⁺ concentration. These results indicate that the apparent increase in insulin sensitivity observed in association with inhibition of Kv1.3 channel activity is mediated by an increase in GLUT4 protein at the plasma membrane, which occurs largely through a Ca²⁺-dependent process. insulin; glucose; diabetes; calcium     

AKT和PTEN在骨肉瘤中表达的相关研究

目的探讨骨肉瘤中AKT和PTEN表达的意义及其临床价值。方法采用免疫组织化学SABC法检测48例骨肉瘤、15例骨软骨瘤标本中AKT、PTEN表达的阳性细胞。结果AKT的阳性表达率在骨肉瘤中为87.5%,在骨软骨瘤中为33.3%,两者比较有显著差异(P<0.05);PTEN的阳性表达率在骨肉瘤中为41.7%,在骨软骨瘤中为86.7%,两者差异显著(P<0.01);在骨肉瘤中,AKT与PTEN呈负相关(r=-0.453,P<0.05)。AKT蛋白表达在有肿瘤复发、肺转移的骨肉瘤中的表达高于无复发及无肺转移的骨肉瘤(P<0.05),而PTEN则相反(P<0.05)。两者的表达均与年龄、性别、肿瘤的大小无关(P>0.05)。结论AKT蛋白的表达在骨肉瘤发生发展中起重要作用,PTEN则可抑制这一作用过程。     

Large-scale analysis of small RNAs derived from traditional Chinese herbs in human tissues

Plant-derived microRNAs have recently been reported to function in human blood and tissues. Controversy was immediately raised due to possible contamination and the lack of large sample sizes. Here, we report thousands of unique small RNAs derived from traditional Chinese medicine(TCM) herbs found in human blood cells and mouse lung tissues using a large-scale analysis.We extracted small RNAs from decoctions of 10 TCM plants(Ban Zhi Lian, Chai Hu, Chuan Xin Lian, Di Ding Zi Jin, Huang Qin, Jin Yin Hua, Lian Qiao, Pu Gong Ying, Xia Ku Cao, and Yu Xing Cao) and obtained millions of RNA sequences from each herb. We also obtained RNA-Seq data from the blood cells of humans who consumed herbal decoctions and from the lung tissues of mice administered RNAs from herbal decoctions via oral gavage. We identified thousands of unique small RNA sequences in human blood cells and mouse lung tissues. Some of these identified small RNAs from Chuan Xin Lian and Hong Jing Tian could be mapped to the genomes of the herbs, confirming their TCM plant origin. Small RNAs derived from herbs regulate mammalian gene expression in a sequence-specific manner, and thus are a superior novel class of herbal drug components that hold great potential as oral gene-targeted therapeutics, highlighting the important role of herbgenomics in their development.     

Arf6 regulates EGF-induced internalization of E-cadherin in breast cancer cells

E-cadherin internalization facilitates dissolution of adherens junctions and promotes tumor cell epithelial-mesenchymal transition (EMT) and migration. Our previous results have shown that Arf6 exerts pro-migratory action in breast cancer cells after EGF stimulation. Despite the fact that EGF signaling stimulates EMT of breast cancer cells, the effect of Arf6 on internalization of E-cadherin of breast cancer cells under EGF treatment remains to be determined. Here, we showed that EGF dose-dependently stimulated E-cadherin internalization by MCF-7 cells with the maximal effect at 50 ng/ml. Meanwhile, EGF treatment markedly increased Arf6 activation. Arf6 was involved in complexes of E-cadherin, and more E-cadherin was pulled down with Arf6 when the activity of the latter was increased. Immunoblotting and immunofluorescence assays showed that transfection breast cancer cells with Arf6-T27N or Arf6 siRNA suppressed EGF-induced E-cadherin internalization. Taken together, our study demonstrated that Arf6 activation plays a potential role in EGF-induced E-cadherin internalization, providing new mechanism underlying the effect of Arf6 on promoting breast cancer cell metastasis.

Electronic supplementary material

The online version of this article (doi:10.1186/s12935-015-0159-3) contains supplementary material, which is available to authorized users.     

基于Maxent模型的粤港澳大湾区水鸟多样性热点研究

粤港澳大湾区拥有丰富的滨海湿地和水网湿地,是水鸟迁徙路线上的重要栖息地。基于55种水鸟的分布数据和15个环境变量数据,利用最大熵模型（Maximum Entropy model）分析了粤港澳大湾区水鸟多样性的空间分布格局及热点地区。研究表明：影响水鸟潜在分布的主要环境因子依次是降雨量季节变化、土地利用类型、距离水源的距离,水鸟丰富度最高出现在粤港澳大湾区的东南区域,呈现往内陆地区逐渐降低的趋势,与当地水资源空间分布一致。水鸟多样性热点地区分布在东南部沿海一带,集中在珠海、江门、深圳、中山等市。研究结果能为粤港澳大湾区水鸟多样性保护规划提供参考,对维持该区域生态系统安全和稳定具有重要意义。