Minimally modified low-density lipoprotein up-regulates endothelin receptors in mouse mesenteric artery through p38 MAPK pathway

AIMTo investigate whether minimally modified low-density lipoprotein (mmLDL) affects the quantity and activity of endothelin （ET） type A (ET_A) and type B (ET_B) receptors in mouse mesenteric artery by activating p38 mitogen-activated protein kinase (MAPK) inflammatory pathway. METHODSThe KM mice were divided into normal saline (NS) group (injection of NS via caudal vein), mmLDL group (injection of mmLDL via caudal vein), LDL group (injection of LDL via caudal vein), mmLDL+SB 203580 group (injection of mmLDL via caudal vein and intraperitoneal injection of p38 MAPK pathway specific inhibitor SB 203580) and mmLDL+DMSO group (injection of mmLDL via caudal vein and intraperitoneal injection of DMSO). Mesenteric artery ring segment vasoconstriction dose-response curves affected by sarafotoxin 6c (S6c) and ET-1 were recorded by the myography system. The mRNA levels of ET_B receptor, ET_A receptor and interleukin-6 (IL-6) were detected by RT-qPCR. The protein levels of ET_B receptor, ET_A receptor, IL-6, p38 MAPK, p-p38 MAPK, NF-κB and p-NF-κB were determined by Western blot. The serum concentration of IL-6 was measured by ELISA. RESULTSThe contractile responses of the blood vessel segments to S6c and ET-1 were significantly increased by mmLDL (P<0.01). The mRNA and protein expression levels of ET_A receptor, ET_B receptor, and IL-6 significantly increased (P<0.01). The protein levels of p-p38 MAPK and p-NF-κB were significantly increased (P<0.01). The serum level of IL-6 was significantly increased (P<0.01). These effects of mmLDL were inhibited by p38 MAPK inhibitor SB 203580. CONCLUSION mmLDL increses the serum concentration of IL-6, up-regulates the expression of IL-6, ET_A receptor and ET_B receptor in mouse mesenteric artery, and enhances the vasoconstriction function medi?ated by ET_A and ET_B receptors, which is related to the activation of p38 MAPK inflammatory pathway and downstream NF-κB pathway.     

Roles of protein phosphatase 4 in down-regulation of eNOS Ser633 phosphorylation induced by palmitic acid in human umbilical vein endotheli?al cells

AIMTo investigate the roles of protein phosphatase 4 (PP4) in down-regulation of endothelial nitric oxide synthase (eNOS) Ser633 phosphorylation induced by palmitic acid (PA). METHODSHuman umbilical vein endothelial cells (HUVECs) were treated with PA at 25 μmol/L, 50 μmol/L, 100 μmol/L and 200μmol/L for 36 h, or treated with PA at 100 μmol/L for 12 h, 24 h, 36 h and 48 h. Protein phosphatase 2A (PP2A) family inhibitor fostriecin (FST, 20 nmol/L) or okadaic acid (OA, 5 nmol/L) was selected to pretreat the HUVECs for 30 min. Protein phosphatase 4 catalytic subunit (PP4c) siRNA or protein phosphatase 2A catalytic subunit (PP2Ac) siRNA was transfected into the HUVECs. The protein expression levels of of eNOS, PP4c and PP2Ac, as well as the level of eNOS Ser633 phosphorylation, were detected by Western blot. The intracellular nitric oxide (NO) content was measured by DAF-FM DA. RESULTS(1) Compared with control group, the levels of eNOS Ser633 phosphorylation were decreased in PA groups in which the HUVECs were treated with 25 μmol/L, 50 μmol/L, 100 μmol/L and 200 μmol/L PA for 36 h (P<0.05) and 100 μmol/L PA for 24 h, 36 h and 48 h (P<0.05). No significant difference in the level of total eNOS protein expression among all the groups was observed. (2) Compared with control group, both FST and OA pretreatment reversed the reduction of eNOS Ser633 phosphorylation (P<0.05) and the decrease in intracellular NO content (P<0.05) induced by PA. No significant difference in the level of total eNOS protein expression among all the groups was observed. (3) Compared with si-Control group, the PP4c protein expression was significantly reduced (P<0.05), while the level of eNOS Ser633 phosphorylation was significantly increased in si-PP4c group (P<0.05). Although the levels of PP2Ac protein expression declined significantly (P<0.05), the level of eNOS Ser633 phosphorylation remained unchanged in si-PP2Ac group. No significant differencein the level of total eNOS protein expression among all the groups was found. CONCLUSION PA significantly reduces the level of eNOS Ser633 phosphorylation and the content of NO in the HUVECs, which may be due to PA inducing the activation of the PP2A family member PP4 rather than PP2A.     

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.     

Role of IP3R in LH-EGFR-induced mouse oocyte meiotic resumption

AIM: To explore the effect of inositol 1, 4, 5-trisposphate receptor (IP₃R) in luteinizing hormone-epidermal growth factor receptor (LH-EGFR)-induced oocyte meiotic resumption. METHODS: Models of mouse cumulus-oocyte complexs (COCs) culture and follicle culture in vitro were generated to study the effects of 2-aminoethyl diphenyl borate (2-APB) and heparin (IP₃R specific inhibitors) on LH/EGF-induced oocyte meiotic resumption and EGF-induced cumulus cell expansion. Real-time PCR was used to detect the mRNA expression of cumulus expansion-related factors. The changes of the intracellular calcium level were monitored using Fluo 3-AM, and the cGMP level was measured by ELISA. RESULTS: The inhibitors of IP₃R, 2-APB and heparin, dramatically reversed EGF-induced oocyte maturation (P<0.05) and decreased cGMP levels in COCs (P<0.05). In addition, 2-APB and heparin reversed EGF-induced cumulus expansion, and significantly inhibited EGF-induced cumulus expansion-related factor expression (P<0.05). The activation of IP₃R increased intracellular calcium level, and the study found that 2-APB and heparin dramatically reversed EGF-induced elevation of calcium level in cumulus cells (P<0.05). Follicular culture in vitro showed that 2-APB and heparin significantly reversed the LH-induced oocyte maturation (P<0.05). CONCLUSION: LH-EGFR signaling pathway increases calcium level in cumulus cells through IP₃R, resulting in meiotic resumption.     

聚对氯苯胺修饰电极的制作及应用研究

通过对聚对氯苯胺膜修饰电极的制作和实验特性的研究,发现聚对氯苯胺膜表面均匀、光滑、牢固,在pH3～11范围内均有响应,测定的重现性好。电极使用长久且稳定性高。该电极和参比电极可直接插入家兔等活体内测定血乳酸的变化情况,对生命科学的活体检测和诊断有其实用意义。     

嗜水气单胞菌UDP-乙酰葡萄糖胺-4-差向异构酶基因的克隆表达及酶的性质

通过水生动物源性嗜水气单胞菌（Aeromonas hydrophila,Ah）强毒株J-1株与弱毒株MR-1株抑制差减杂交得到gneJ差减片段,该片段与创伤弧菌（Vibrio vulnificus）和人源嗜水气单胞菌的UDP-乙酰葡萄糖胺-4差向异构酶基因有较高的同源性。扩增完整的gneJ基因,进行分布检测并将其克隆至质粒pET32a（＋）,在大肠杆菌（Escherichia coli）BL21中进行异源表达,SDS-PAGE结果显示,重组表达蛋白的分子量约59．7kD,与理论推测值基本相同。酶活性分析表明,该重组蛋白可将UDP-乙酰葡萄糖胺转化为UDP-乙酰半乳糖胺,确证gneJ基因编码蛋白为UDP-乙酰葡萄糖胺-4-差向异构酶。Western Blot分析显示,嗜水气单胞菌J-1株胞外产物的兔抗血清可识别该重组蛋白,表明该蛋白与天然蛋白有相同的抗原性。以重组蛋白为免疫原,对小鼠进行动物保护实验,结果显示,该重组蛋白对小鼠有60％的保护率,证明UDP-乙酰葡萄糖胺-4差向异构酶可作为亚单位疫苗的侯选成分。     

昆明市马铃薯新品种-昆薯4号

昆薯4号属中晚熟品种,是昆明市农业科学研究院从国际马铃薯中心（cIP）引进的马铃薯选育材料中筛选而来,其杂交组合母本：381379．9,父本：XY16。在品种比较试验和云南省马铃薯品种区域试验及生产试验中表现突出,2009年该品种通过云南省农作物品种审定委员会审定,命名为“昆薯4号”。     

虎杖提取物对CCl4诱导建鲤损伤肝细胞生化指标的影响

为探明虎杖对鱼类损伤肝细胞是否具有修复作用,分离建鲤肝细胞,设6个试验组：I组(空白对照组);II组(CCl4模型组);III组(800 μg/mL虎杖提取物对照组);IV组(造模前200、400、800 μg/mL虎杖提取物处理组);V组(造模后200、400、800 μg/mL虎杖提取物处理组);VI组(造模前、后200、400、800 μg/mL虎杖提取物处理组)。各组细胞经处理后继续培养12 h,收集肝细胞培养液,检测上清培养液中丙氨酸转氨酶(GPT)、天冬氨酸转氨酶(GOT)、乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)等酶活性,丙二醛(MDA)含量及肝细胞的存活率。结果表明：III组肝细胞培养液中GOT、GPT、LDH、SOD、MDA值及细胞存活率与I组相比无明显变化(P＞0.05),说明800 μg/mL虎杖提取物没有细胞毒性,可用于后续试验;与II组相比,IV组中800 μg/mL虎杖提取物处理组的效果优于其他2个浓度组,可以极显著降低培养液中GOT值(P<0.01),显著降低培养液中GPT、LDH值(P<0.05),显著提高肝细胞的存活率(P<0.05),而MDA含量和SOD值差异无统计学意义(P＞0.05);与II组相比,V组中400 μg/mL虎杖提取物效果优于其他2个浓度组,可以极显著降低培养液中GOT值(P<0.01),显著降低培养液中LDH值(P<0.05),显著提高肝细胞的存活率(P<0.05),但GPT值、MDA含量、SOD的差异无统计学意义(P＞0.05);与II组相比,VI组中800 μg/mL虎杖提取物效果优于其他2个浓度组,能极显著降低培养液中GOT、GPT、LDH在肝细胞中的释放(P<0.01),显著降低培养液中MDA含量(P<0.05),显著提高肝细胞的存活率(P<0.05)。综合以上结果,以VI组中800 μg/mL的虎杖提取物效果最好,能有效抑制CCl4所造成的肝细胞损伤,其机制可能与其抗氧化作用有关。     

孔隙结构对水稻土温室气体排放的影响

土壤结构影响水分和气体的运动和土壤生物活动,进而影响稻田温室气体排放.为探明土壤结构对水稻生长过程中温室气体排放的影响,选取江苏宜兴的湖白土和江西进贤的红壤性水稻土进行盆栽试验.设置不搅动(NP)、搅动(PD)和搅动后掰土回填(RP)3个处理.应用X射线CT成像技术分析不同处理土壤孔隙结构,通过静态箱法测定水稻生长过程...     

水氮用量对设施栽培蔬菜地土壤氨挥发损失的影响

【目的】针对我国设施蔬菜生产中存在的水肥过量施用问题,研究不同水氮条件下黄瓜-番茄种植体系内的土壤氨挥发特征,探讨影响设施菜地土壤氨挥发的重要因子,为降低氮肥的氨挥发损失、 建立合理的灌溉和施肥制度提供参考。【方法】以华北平原设施黄瓜-番茄轮作菜地为研究对象,设常规灌溉(W1)和减量灌溉(W2)2个灌溉水平,每种灌溉水平下设不施氮(N0)、 减量施氮(N1)和常规施氮(N2)3个氮水平,共6个处理组合(W1N0、 W1N1、 W1N2、 W2N0、 W2N1、 W2N2)。采用通气法监测不同水氮条件下黄瓜-番茄轮作体系内的土壤氨挥发动态,分析与土壤氨挥发相关的主要影响因子。【结果】设施黄瓜-番茄种植体系内表层(0—10 cm)土壤铵态氮受施肥的影响波动较大,与常规施氮(N2)相比,相同灌水条件下减量施氮(N1)处理的0—10 cm土层铵态氮浓度最高值降低了25.1%~30.3%(P 0.05)。减量施氮可显著降低土壤氨挥发速率。与常规施氮(N2)相比,减量施氮处理(N1)在黄瓜季和番茄季内的氨挥发速率均值分别降低了21.1%~22.8%(P0.05)和16.5%~17.9%(P0.05)。整个黄瓜-番茄轮作周期内,土壤氨挥发损失量和氮肥的氨挥发损失率分别为17.8~48.1 kg/hm2和1.23%~1.44%。与常规施氮(N2)相比,减量施氮处理(N1)的土壤氨挥发损失量及氮肥的氨挥发损失率分别降低了19.3%~20.0%(P0.05)和0.85~0.92个百分点。各处理土壤氨挥发速率与0—10 cm土壤铵态氮浓度呈显著或极显著正相关,说明0—10 cm土壤铵态氮浓度是土壤氨挥发的重要驱动因子。与常规灌溉(W1)相比,减量灌溉(W2)条件下设施菜地土壤氨挥发速率及氨挥发损失量略有增加(P0.05)。适宜减少氮肥及灌溉量不仅能够维持较高的蔬菜产量,而且显著提高了灌溉水和氮肥的利用效率。其中减量施氮处理(N1)的氮肥农学效率比常规施氮(N2)提高了95.4%~146.4%; 减量灌溉(W2)的灌溉水农学效率比常规灌溉(W1)提高了27.7%~54.0%。【结论】通过合理的节水减氮措施可达到抑制氮肥氨挥发损失、 增加产量以及提高水氮利用效率的目的。在供试条件下,节水30%左右、 减施氮量25%的水氮组合(W2N1)具有较佳的经济效益与环境效应。