Involvement of Ca2+/calmodulin-dependent protein kinase II in endothelial NO production and endothelium-dependent relaxation

Nitric oxide (NO) is synthesized from l-arginine by the Ca(2+)/calmodulin-sensitive endothelial NO synthase (NOS) isoform (eNOS). The present study assesses the role of Ca(2+)/calmodulin-dependent protein kinase II (CaMK II) in endothelium-dependent relaxation and NO synthesis. The effects of three CaMK II inhibitors were investigated in endothelium-intact aortic rings of normotensive rats. NO synthesis was assessed by a NO sensor and chemiluminescence in culture medium of cultured porcine aortic endothelial cells stimulated with the Ca(2+) ionophore A23187 and thapsigargin. Rat aortic endothelial NOS activity was measured by the conversion of l-[(3)H]arginine to l-[(3)H]citrulline. Three CaMK II inhibitors, polypeptide 281-302, KN-93, and lavendustin C, attenuated the endothelium-dependent relaxation of endothelium-intact rat aortic rings in response to acetylcholine, A23187, and thapsigargin. None of the CaMK II inhibitors affected the relaxation induced by NO donors. In a porcine aortic endothelial cell line, KN-93 decreased NO synthesis and caused a rightward shift of the concentration-response curves to A23187 and thapsigargin. In rat aortic endothelial cells, KN-93 significantly decreased bradykinin-induced eNOS activity. These results suggest that CaMK II was involved in NO synthesis as a result of Ca(2+)-dependent activation of eNOS.     

携带EBV-LMP2基因的DNA疫苗、腺相关病毒疫苗和腺病毒疫苗免疫小鼠的特异性细胞免疫应答

细胞免疫应答, 尤其是CD3⁺CD8⁺细胞毒性T淋巴细胞(cytotoxic T lymphocyte, CTL), 在控制病毒感染中扮演着重要角色. 鼻咽癌(nasopharyngenl carcinoma, NPC)与EB病毒(epstein-barr virus, EBV)持续感染密切相关, 在鼻咽癌疫苗的研究中, 人们将研究重点放在增强特异性抗病毒CTL应答上. 本研究单独或联合使用表达EB病毒潜伏膜蛋白抗原2(epstein-barr virus latent membrane protein 2, EBV-LMP2)的DNA疫苗、腺相关病毒(adeno-associated virus, AAV)疫苗、非复制5型腺病毒疫苗(Ad5), 分别于0, 2, 4周肌肉注射免疫4~6周龄的雌性Balb/C小鼠, IFN-γ 免疫斑点法检测EBV-LMP2特异性细胞免疫应答水平. 疫苗诱导的特异性细胞应答水平与疫苗的免疫策略有关. 其中, 使用3种载体疫苗联合免疫的效果最好, 其次则是先使用DNA疫苗免疫两次, 再用腺病毒疫苗加强免疫一次的联合免疫方法. DNA疫苗和AAV疫苗单独免疫能够诱导出特异性的CTL, 但与联合免疫相比诱导的应答水平很低. 结果表明, 使用DNA, AAV和腺病毒载体疫苗联合免疫, 能够更好地诱导机体产生特异性细胞免疫应答, 为鼻咽癌的防治提供了很好的疫苗策略.     

Neutrophil apoptosis: A target for enhancing the resolution of inflammation

Neutrophils are essential for host defense and their programmed cell death and removal are critical for the optimal expression as well as for efficient resolution of inflammation. Delayed neutrophil apoptosis or impaired clearance of apoptotic neutrophils by macrophages contributes to the progression of chronic inflammation. Under most conditions, neutrophils are exposed to multiple factors and their fate would ultimately depend on the balance between pro‐survival and pro‐apoptotic signals. Life or death decisions are tightly controlled by a complex network of intracellular signaling pathways. Accumulating data indicate that receptors, such as the formyl peptide receptor 2/lipoxin receptor or β₂‐integrins can generate contrasting cues in neutrophils in a ligand‐specific manner and suggest a hierarchy among these signals. In this article, we review recent advances on how pro‐apoptosis and pro‐survival signals interact to determine the fate of neutrophils and the inflammatory response, and highlight novel pharmacological strategies that could be used to enhance the resolution of inflammation by redirecting neutrophils to apoptosis. J. Cell. Biochem. 108: 1039–1046, 2009. © 2009 Wiley‐Liss, Inc.     

650 nm激光穴位照射与针刺治疗良性前列腺增生症的疗效对比观察

目的:观察、评估650 nm激光照射穴位治疗良性前列腺增生症(中医辨证为肾阳虚)的临床疗效。方法:对30例良性前列腺增生症患者,采用650 nm激光,照射会阴、关元、肾俞,并随机选取30例,用针刺治疗作临床对比观察,针刺肾俞、秩边、关元、命门、足三里、脾俞、三阴交、次髎等穴。结果:激光照射穴位与针刺治疗均对良性前列腺增生症有较好的疗效,两组治疗前后症状评分、尿动力学均有显著的改变(P<0.05),但两者的临床疗效差异无统计学意义(P>0.05)。结论:650 nm激光穴位照射是临床治疗良性前列腺增生症(肾阳虚型)的有效治疗方法之一,值得深入研究。     

非线性种群反应扩散系统奇摄动ROBIN问题

研究了一类具有非线性种群反应扩散系统奇摄动Robin初始边值问题.在适当的条件下,利用微分不等式理论,讨论了问题解的存在性和渐近性态.     

不同倍性香石竹杂交花粉管生长荧光显微观察及结实研究

以香石竹四倍体材料‘紫蝴蝶’(2n=4x=60)为母本,二倍体材料‘珍珠粉’和‘NH6’(2n=2x=30)为父本,利用荧光显微镜观察其授粉后花粉管生长情况,统计其座果率、亲和指数及种子萌发率,并对杂交后代进行倍性鉴定。结果表明,在‘紫蝴蝶’柱头上,‘珍珠粉’和‘NH6’的花粉2h开始萌发,花粉管多处出现胼胝质塞,且花柱组织出现胼胝质反应,4h花粉管到达柱头中部并出现胼胝质塞,6h花粉管到达柱头基部,17h柱头基部的花粉管增多,花粉管进入子房组织且子房组织出现胼胝质反应,17～24h花粉管能与胚珠结合,但结合率低;‘紫蝴蝶’ב珍珠粉’杂交未获得植株,‘紫蝴蝶’בNH6’杂交获得3株植株,染色体倍性鉴定表明3株植株均为四倍体,这可能是‘NH6’产生2n配子的缘故。     

四种植物精油对黑翅土白蚁触杀和驱避作用

为筛选出高效防治黑翅土白蚁的天然植物精油,减少有机合成农药的使用,该文研究了大蒜精油、肉桂油、丁香油和印楝素油四种植物精油对黑翅土白蚁的触杀效果和驱避作用。结果表明:大蒜精油、肉桂油和丁香油的浓度为5和10 mg·m L~(-1)时,处理2 h后,黑翅土白蚁的校正死亡率达100%,而相同浓度的印楝素油和对照处理的黑翅土白蚁校正死亡率低于5%。随着处理时间延长,浓度为1.25和2.5 mg·m L~(-1)的大蒜精油、肉桂油和丁香油处理6 h时,黑翅土白蚁的校正死亡率仍达100%,而此时对应的印楝素油和对照处理的黑翅土白蚁校正死亡率仅为10%,说明大蒜精油、肉桂油和丁香油对黑翅土白蚁具有较强的触杀效果。大蒜精油、丁香油和肉桂油在处理黑翅土白蚁2 h后LC_(50)值(半致死量)分别为1.572、1.05和1.03mg·m L~(-1),说明肉桂油对黑翅土白蚁的毒性相对最大,触杀效果最好。此外,10 mg·m L~(-1)的大蒜精油、肉桂油、丁香油和印楝素油的驱避试验表明,处理4、6、8和12 h后,大蒜精油、肉桂油和丁香油三精油处理区的黑翅土白蚁数均显著低于对照区的,驱避率总体93%,而对应的印楝素油的驱避率总体28.5%,表明大蒜精油、丁香油和肉桂油三种植物精油对黑翅土白蚁均有显著的驱避活性。综上可知,四种植物精油中大蒜精油、肉桂油和丁香油在防治黑翅土白蚁方面应用潜力很好,是开发绿色环保白蚁防治药剂的可选材料。     

人类巨细胞病毒编码US28受体拮抗性多肽的筛选鉴定及体外活性研究

立足US28的广谱趋化因子结合活性,寻找拮抗剂多肽的先导物．通过预测US28的结合模拟位设计多肽序列,合成相关多肽,再利用自建的25种趋化因子随机构成的噬菌体12肽库筛选结合模拟位,竞争性ELISA方法验证,从而获得30个阳性克隆,测序并推导氨基酸序列,得到7种序列：GSESLNAHCALW、EIDGFNAHCALL、VIARLNAHCALR、ATECLNAHCALW、VIESLNAHCALW、DNGSINAHCALL及VKKTLNAHCALR．所有序列富含疏水氨基酸,其中8个克隆有LNAHCAL保守序列．采用趋化抑制实验检测多肽对生理性趋化因子引起的细胞迁移活性的影响,流式细胞仪检测细胞内钙离子浓度变化．结果表明,利用人源趋化因子序列构建肽库并筛选有效序列的方法取得成功,筛选出的阳性克隆保守序列LNAHCAL可以模拟Human MIP-1β与US28 N端的结合位,从而与合成多肽H22结合,多肽H22可以阻断受体结合生理性趋化因子形成的趋化作用,本身不引起趋化运动,不影响胞内信号转导和细胞自然活性,具有广谱趋化因子拮抗剂的可能性．     

Nitrogen addition reduces soil respiration in a mature tropical forest in southern China

Response of soil respiration (CO₂ emission) to simulated nitrogen (N) deposition in a mature tropical forest in southern China was studied from October 2005 to September 2006. The objective was to test the hypothesis that N addition would reduce soil respiration in N saturated tropical forests. Static chamber and gas chromatography techniques were used to quantify the soil respiration, following four‐levels of N treatments (Control, no N addition; Low‐N, 5 g N m^?2 yr^?1; Medium‐N, 10 g N m^?2 yr^?1; and High‐N, 15 g N m^?2 yr^?1 experimental inputs), which had been applied for 26 months before and continued throughout the respiration measurement period. Results showed that soil respiration exhibited a strong seasonal pattern, with the highest rates found in the warm and wet growing season (April–September) and the lowest rates in the dry dormant season (December–February). Soil respiration rates showed a significant positive exponential relationship with soil temperature, whereas soil moisture only affect soil respiration at dry conditions in the dormant season. Annual accumulative soil respiration was 601±30 g CO₂‐C m^?2 yr^?1 in the Controls. Annual mean soil respiration rate in the Control, Low‐N and Medium‐N treatments (69±3, 72±3 and 63±1 mg CO₂‐C m^?2 h^?1, respectively) did not differ significantly, whereas it was 14% lower in the High‐N treatment (58±3 mg CO₂‐C m^?2 h^?1) compared with the Control treatment, also the temperature sensitivity of respiration, Q₁₀ was reduced from 2.6 in the Control with 2.2 in the High‐N treatment. The decrease in soil respiration occurred in the warm and wet growing season and were correlated with a decrease in soil microbial activities and in fine root biomass in the N‐treated plots. Our results suggest that response of soil respiration to atmospheric N deposition in tropical forests is a decline, but it may vary depending on the rate of N deposition.     

葡激酶N端突变体的分子设计,高效表达与分离纯化

同源模建人微小纤溶酶原（ｍｉｃｒｏｐｌａｓｍｉｎｏｇｅｎ ,ｍｐｌｇ）的三维结构,建立葡激酶（ｓｔａｐｈｙｌｏｋｉｎａｓｅ,Ｓａｋ）,ｍｐｌｇ计算机分子对接的结构模型,模型与已有的实验结果基本相符。为研究葡激酶Ｎ端结构与功能的关系,进一步改造葡激酶分子,根据复合物结构模型设计了Ｎ端缺失１５个氨基酸的葡激酶突变体。