Sulodexide protects against hyperglycemia-induced retinal vascular injury via suppressing NOX4/NLRP3 pathway

AIM To investigate the effect of sulodexide (SDX) on high glucose-induced damage in retinal microvascular endothelial cells. METHODS (1) High-fat diet combined with intraperitoneal injection of streptozocin were used to induce type 2 diabetes mellitus (DM) followed by injection of saline or SDX in C57BL/6J male mice. Retinal microvascular leakage and density, and the protein levels of NLRP3 inflammasome-related proteins, zonula occludens-1 (ZO-1) and NADPH oxidase 4 (NOX4) were measured. (2) Human retinal microvascular endothelial cells (HRMECs) were treated with normal glucose or high glucose with or without SDX, and were further transfected with siRNA to knock down NOX4, or infected by adenovirus to over-express NOX4. The protein levels of ZO-1, VE-cadherin (VE-Cad), NOX4 and NLRP3 inflammasome-related proteins as well as the level of reactive oxygen species (ROS) were detected. RESULTS Treatment with SDX increased the protein level of ZO-1, attenuated retinal leakage and NLRP3 inflammasome activation, and enhanced the density of microvasculature and the number of ganglion cells in diabetic retinas. The protein levels of ZO-1 and VE-Cad were decreased, while the levels of NOX4, NLRP3 inflammasome-related proteins and ROS generation were increased in high glucose-treated HRMECs. Silencing of NOX4 inhibited high glucose-induced increases in NLRP3 inflammasome and ROS generation, and decreases in the protein levels of ZO-1 and VE-Cad. Over-expression of NOX4 significantly increased the levels of NLRP3 inflammasome-related proteins and ROS generation in HRMECs, and reduced the protein levels of ZO-1 and VE-Cad. Treatment with SDX partly reversed NOX4 over-expression-induced changes. CONCLUSION SDX alleviates hyperglycemia-induced retinal microvascular endothelial injury via inhibiting NOX4/ROS/NLRP3 pathways.     

Mechanism of elemene enhancing radiosensitivity of human glioma U251 cells

AIM To investigate the effect of elemene on the radiosensitivity of human glioma U251 cells and its mechanism. METHODS The U251 cells were used as a glioma model in vitro, and were exposed to different concentrations of elemene and different doses of radiation. The cell viability was measured by MTT assay, the apoptosis and cell cycle distribution were analyzed by flow cytometry, and the related protein levels were determined by Western blot. RESULTS Elemene inhibited the viability of U251 cells in vitro and enhanced the radiosensitivity of the cells. The cells in radiotherapy combined with elemene group had higher rates of early apoptosis, secondary necrosis and total cell death than those in radiation group. Elemene induced G₂/M phase arrest in the U251 cells. Elemene reduced the protein expression of cell division cycle protein 2 （Cdc2）, which resulted in the decrease in cyclin B1 expression induced by radiotherapy, thereby inhibiting the formation of cyclin B-Cdc2 complex. Elemene reduced Cdc2 activity by inhibiting the phosphorylation of Cdc2 protein at threonine 161, thereby inducing G₂/M phase arrest in the cells. It also mediated apoptosis by down-regulating survivin expression. CONCLUSION Elemene may increase the sensitivity of U251 cells to radiotherapy by down-regulating Cdc2 protein, decreasing cyclin B1 expression, inhibiting the formation of cylcin B-Cdc2 complex and down-regulating the expression of survivin.     

Importance of mitochondrial calcium uniporter in high glucose-induced cardiac myocyte apoptosis

AIM: To investigate the role of mitochondrial calcium uniporter (MCU) in high glucose(HG)-induced apoptosis of cardiac myocytes. METHODS: Cardiac myocytes were exposed to normal glucose (5.5 mmol/L glucose+ 19.5 mmol/L mannitol), HG (25 mmol/L glucose), or HG combined with 5 μmol/L spermine for 72 h. Mitochondrial free Ca²⁺ concentration ([Ca²⁺]_m), MCU at mRNA and protein levels, pyruvate dehydrogenase (PDH) activity, mitochondrial membrane potential (Δψ_m), the levels of ATP and reactive oxygen species (ROS), and apoptosis were determined. RESULTS: The [Ca²⁺]_m, the mRNA and protein levels of MCU, PDH activity, ATP levels, and Δψ_m were reduced (P<0.05), while ROS content and the protein levels of caspase-9 and caspase-3 were increased in HG group (P<0.05). Adding 5 μmol/L spermine returned these parameters toward control levels (P<0.05). Moreover, apoptosis was reduced by adding spermine and HG treatment (P<0.05). CONCLUSION: HG-induced cardiac myocyte apoptosis may be associated with the decreased MCU expression and activity, abnormal mitochondrial Ca²⁺ handling, deviant mitochon-drial respiratory chain, and mitochondrial dysfunction.     

Molecular mechanisms of Belinostat inhibiting viability of osteosarcoma cells

AIM: To observe the effect of histone deacetylase inhibitor (HDACi) Belinostat on the viability of osteosarcoma cells and to study the underlying mechanism. METHODS: Osteosarcoma cell lines SAOS-2 and U2OS were incubated with Belinostat at different concentrations in vitro. The viability of the cells was measured by MTT assay. The activity of caspase-3/-7 and the DNA fragmentation were detected by fluorescence probe and ELISA, respectively. Western blot was used to detect the levels of histone acetylation, expression of PTEN, caspase-3, Bcl-xL and Akt, and phosphorylation of glycogen synthetase kinase 3β (GSK-3β) and Akt. Finally, the cells were incubated with Belinostat and doxorubicin at different concentrations, and then the combination index (CI) was calculated by MTT. RESULTS: Belinostat at 0.5, 1, 2.5 and 5 μmol/L inhibited the viability of U2OS cells and SAOS-2 cells in a dose-dependent manner, induced DNA fragmentation, enhanced caspase-3/-7 activity, and promoted the activation of caspase-3. At the same time, in the SAOS-2 cells, the expression of Bcl-xL was reduced, and the acetylation of histones H3 and H4 was increased. The results of Western blot showed that phosphorylation levels of Akt and GSK-3β in U2OS cells and SAOS-2 cells were decreased significantly after treatment with Belinostat (P<0.05). MTT results showed that combination of Belinostat and doxorubicin further reduced the viability of U2OS and SAOS-2 cells (CI<1). CONCLUSION: Belinostat inhibits the viability of osteosarcoma cells treated with doxorubicin, and the mechanism may be related to the inhibition of Akt signaling pathway.     

LED补光组合对大棚越橘生长发育的影响

以2年生南高丛越橘‘Emerald’为材料，以大棚内自然光作为对照，研究LED光源的红蓝光（3︰1和6︰1）、紫外光（UVA）对植株长势、叶片光合作用、碳氮代谢、开花基因表达及开花率、果实品质的影响。结果表明：红蓝光组合处理下‘Emerald’的植株营养生长较旺盛，株高、1年生枝条长度和粗度显著高于对照。此外，红蓝光（6︰1）处理下叶片的叶绿素相对含量、比叶重和净光合速率均显著提高。红蓝光组合也可诱导植株开花，开花基因FT表达量和开花率明显高于对照。紫外光下叶片的氮含量、开花率及FT基因表达量显著高于对照。不同光质组合补光对‘Emerald’的果实品质有显著影响，红蓝光（3︰1）处理下果实的质量、横纵径、可溶性固形物、可溶性糖、花青素含量和糖酸比均高于对照。总之，不同光质补光会促进越橘‘Emerald’的生长发育，红蓝光6︰1组合对促进营养生长作用相对较大，红蓝光3︰1组合对提高果实品质效果较好。紫外光虽能改变植株形态和促进开花，但果实品质提高效果较红蓝光处理稍弱。     

晚熟苹果新品种‘华蜜’

苹果新品种‘华蜜’是以‘金冠’作母本，‘华富’作父本，杂交选育而成。果实圆锥形，果形指数1.1，平均单果质量220 g；果皮鲜红色，底色黄绿色，果面较光滑，果点中大；果肉较细，淡黄色，肉质松脆多汁，去皮硬度7.5 kg • cm-2，可溶性固形物含量17.5%，口感香甜，有香味，品质上等。果实生育期168 d左右，属晚熟品种，在辽宁省兴城地区10月下旬成熟，产量32 t • hm-2。     

大粒板栗新品种‘秦栗2号’

（西北农林科技大学林学院，陕西杨凌 712100）     

黄瓜耐盐性与耐盐相关基因的研究进展

黄瓜是重要的设施蔬菜之一,由于其根系具有脆弱、好气、分布较浅的特点,对盐渍环境适应性较差,土壤盐渍化已成为影响其产量和品质的主要限制因素。本文主要从光合作用、离子毒害、细胞膜和抗氧化酶等角度阐述了黄瓜对盐胁迫的反应及其基本的耐盐机理,介绍了黄瓜耐盐(包括与Na~+/K~+、植物激素和活性氧的调节)相关基因的研究进展,并指出了黄瓜耐盐性研究中存在的问题,旨在为揭示黄瓜的耐盐机理与耐盐育种提供理论参考。     

Comparative analysis of genetic effects of wheat‐Dasypyrum villosum translocations T6V#2S·6AL and T6V#4S·6DL

Wheat‐Dasypyrum villosum translocations T6V#2S·6AL and T6V#4S·6DL, carriers of Pm21 and PmV, respectively, confer high resistance to wheat powdery mildew. For better understanding of the difference in genetic effect between them, a RIL population was constructed based on the cross between “Yangmai 18” carrying T6V#2S·6AL and “Yangmai 22” carrying T6V#4S·6DL. Analysis of distribution of the translocations showed that T6V#2S·6AL is much more transmittable than T6V#4S·6DL. By comparing their effects on main agronomic traits, we firstly found that T6V#2S·6AL contributes greatly to top spikelet fecundity, but causes a decrease of 6.7%–10.5% of spike number. No stable effects of T6V#4S·6DL on agronomic traits were found, except for positive effect on plant height. Excitingly, a new recombinant, T6V#4S‐6V#2S·6AL carrying PmV, was screened and proved to have a higher transmission rate than the original translocation T6V#4S·6DL, which will greatly promote the utilization of PmV. The above conclusions of this research will provide important guidance for utilization of Pm21 and PmV more effectively, in wheat powdery mildew resistance breeding.