单纯疱疹病毒Ⅱ型潜伏、激活细胞模型建立

【目的】建立单纯疱疹病毒Ⅱ型(HSV-2)潜伏感染人神经母细胞瘤细胞株SH-SY5Y及激活的细胞模型。【方法】分别加入20,40,60,80,100,120,140μmol/L的阿昔洛韦(ACV),观察对SH-SY5Y细胞生物性状的影响;在ACV存在的情况下,分别将含有0.1、1、10、100MOI的病毒液接种SH-SY5Y细胞,运用相差显微镜观察病毒对细胞的影响,确定潜伏的建立;分别用41℃、42℃、43℃、44℃、45℃加热0.5h、1.0h、1.5h、2.0h、2.5h,观察加热时间及温度诱导HSV-2在SH-SY5Y细胞中激发的最适条件;加入25、50、75、100、125μmol/L福斯高林(Forskolin)诱导病毒在细胞中激活,探讨诱导的最佳浓度;对HSV-2在SH-SY5Y细胞中的潜伏及激发进行验证并测序;运用相差显微镜观察病毒激活后细胞形态的变化。【结果】60μmol/LACV的存在最适合HSV-2在SH-SY5Y细胞中建立潜伏状态;1-10MOI的感染量均能取得较好的病毒潜伏及激发效果;通过观察,病毒在SH-SY5Y细胞中最长可潜伏14d;43℃,1.5h及75μmol/LForskolin均为诱导病毒潜伏激发的最佳条件;相差显微镜观察病毒激发后细胞病变,从24h到72h,细胞变性、坏死的程度、数量随感染时间延长而增加;HSV-2LAT、gG基因PCR扩增及电泳结果,证实病毒在细胞中的潜伏及激活。【结论】初步在人神经母细胞瘤细胞株SH-SY5Y上建立了HSV-2潜伏感染及激活的细胞模型,为下一步研究HSV-2的潜伏与激发机理,了解HSV-2的致病机制打下基础。     

Identification of an anticoagulant peptide that inhibits both fXIa and fVIIa/tissue factor from the blood-feeding nematode Ancylostoma caninum

Factor VIIa-tissue factor complex (fVIIa/TF) and factor XIa (fXIa) play important roles in the initiation and amplification of coagulation, respectively. They may be good targets for the development of novel anticoagulants to treat and prevent thromboembolic disease. In this study, we cloned, expressed and identified a novel anticoagulant peptide, AcaNAP10, from the blood-feeding nematode Ancylostoma caninum. AcaNAP10 showed potent anticoagulant activity and doubled the activated partial thromboplastin and prothrombin times at estimated concentrations of 92.9 nM and 28.8 nM, respectively. AcaNAP10 demonstrated distinct mechanisms of action compared with known anticoagulants. It inhibited fXIa and fVIIa/TF with IC₅₀ values of 25.76 ± 1.06 nM and 123.9 ± 1.71 nM, respectively. This is the first report on an anticoagulant that can inhibit both fXIa and fVIIa/TF. This anticoagulant peptide may be an alternative molecule for the development of novel anticoagulants.     

冠心病患者血清CRP的变化及其临床意义

目的:探讨冠心病(CAD)患者血清C-反应蛋白(CRP)的变化及其临床意义.方法:运用全自动生化分析仪检测97例CAD患者和28例对照组人群的血清CRP,将CAD患者进一步分为稳定性心绞痛组(SAP)、不稳定心绞痛(UAP)、心肌梗死组(AMI)三组.另外,将97例CAD惠者分为1支病变组、2支病变组和3支病变组.所有研究对象均行选择性冠状动脉造影检查.结果:(1)与对照组比较,CAD患者血清CRP显著升高(P<0.05);(2)与对照组比较,SAP组、UAP组和AMI组患者血清CRP均显著升高(P<0.05);与SAP组比较,UAP组和AMI组患者血清CRP亦显著升高(P<0.05);(3)与对照组比较,1支病变组、2支病变组和3支病变组患者血清CRP均显著升高(P<0.05);与1支病变组比较,2支病变组和3支病变组患者血清CRP亦显著升高(P<0.05);与2支病变组比较,3支病变组患者血清CRP亦显著升高(P<0.05).结论:(1)CRP可能与CAD有关;(2)CRP异常升高可能反映冠状动脉斑块的不稳定和CAD的严重程度.     

Ⅱ期经皮输尿管镜治疗输尿管上段结石梗阻致孤立肾急性肾功能衰竭

目的:探讨经皮肾微造瘘术后Ⅱ期经皮输尿管镜治疗输尿管上段结石致孤立肾急性肾功能衰竭的安全性与有效性。方法:从2004年7月～2009年5月,利用经皮肾微造瘘建立经皮肾通道,引流1周后肾功能明显好转,再行经皮肾输尿管镜碎石治疗孤立肾输尿管上段结石。结果:16例患者中,所有患者均为单通道取石,结石清除率例(81.2%),未出现高热、出血等并发症。术后1月复查13例无结石残留。结论:微创经皮输尿管镜分期治疗输尿管上段结石致孤立肾急性肾功能衰竭是安全、有效的,同传统经皮肾镜相比,具有对病人创伤小,易恢复等优点。     

A potential tocopherol acetate loaded palm oil esters-in-water nanoemulsions for nanocosmeceuticals

Background  

Cosmeceuticals are cosmetic-pharmaceutical hybrids intended to enhance health and beauty of the skin. Nanocosmeceuticals use nano-sized system for the delivery of active ingredients to the targeted cells for better penetration. In this work, nanoemulsion from palm oil esters was developed as a delivery system to produce nanocosmeceuticals. The stability of the resulting formulation was tested using various methods. In addition, the effect of components i.e. Vitamin E and Pluronic F-68 on the formulation was also studied.     

野生甜瓜'云甜-930'对白粉病抗性的遗传分析

以经过多代自交选育的高抗白粉病材料野生甜瓜'云甜-930'(P1)与感病栽培甜瓜'华莱士'(P2)组配,构建了P1、P2、F1、B1、B2和F2 6个世代,对大棚栽培条件下各世代材料的白粉病自然发病状况进行考察,用植物数量性状主基因+多基因混合遗传模型的多世代联合分析法,研究野生甜瓜'云甜-930'对白粉病的抗性遗传规律.结果显示:野生甜瓜'云甜-930'对白粉病的抗性遗传符合2对加性-显性-上位性主基因+加性-显性-上位性多基因混合遗传模型(E-0),2对主基因的加性效应相等,da和db均为-15.76,2对主基因的显性效应ha和hb分别为14.98和19.87,第2对主基因的显性效应大于第1对;2对主基因互作效应中除了显性×显性效应(l=-7.73)较小外,其它互作效应均较大,加性×加性效应i为31.46,加性×显性效应jab为-26.86,而显性×加性效应jba为-17.07.该组合的B1、B2和F2群体抗病性主基因遗传率分别为73.31%、69.15%和97.61%,多基因遗传率分别为18.83%、25.86%和0,环境变异在2.39%～7.86%之间.研究表明,野生甜瓜'云甜-930'对白粉病的抗性受2对加性-显性-上位性主基因+加性-显性-上位性多基因控制,同时还受到环境的影响.在甜瓜抗白粉病育种中,在F2代主基因的选择效率最高.     

益生菌ZD02的分离及其在对虾集约化精养中的应用

从养殖环境分离纯化芽孢杆菌6株,对其进行消化酶活检测,根据检测的酶活指标选择ZD02作为待试验菌株并进行鉴定和热稳定性检测。结果表明:ZD02被鉴定为枯草芽孢杆菌,具有耐受高温的特性,90℃水浴10min存活率93%,95℃水浴5min存活率93%,100℃水浴5min存活82%,95℃和100℃水浴2min不影响芽孢菌存活,在调质温度102℃条件下,制粒后芽孢菌存活率94%;将经过鉴定的枯草芽孢杆菌发酵培养后应用于凡纳滨对虾(Litopenaeus vannamei)的集约化精养殖,试验组A在基础饲料中添加0.3%的芽孢杆菌。试验组B保持配方成本不变的基础上添加0.3%的芽孢杆菌,结果显示,试验组的成活率显著高于对照组(P0.05),试验组A、B的产量分别比对照组提高了43.0%和36.7%(P0.05),产值分别提高35.4%和21.0%(P0.05),饲料系数比对照组分别降低了9.9%和8.7%(P0.05);换水耗电、水质调节剂、内服药物3项费用合计,A组和B组较对照组分别节省了21.3%和22.3%(P0.05);实验组之间各项指标差异不显著(P0.05);表明,在对虾的集约化精养过程中合理利用益生芽孢杆菌可以增产增收。     

Multiphysics simulation of blood flow and LDL transport in a porohyperelastic arterial wall model

Atherosclerosis localizes at a bend andor bifurcation of an artery, and low density lipoproteins (LDL) accumulate in the intima. Hemodynamic factors are known to affect this localization and LDL accumulation, but the details of the process remain unknown. It is thought that the LDL concentration will be affected by the filtration flow, and that the velocity of this flow will be affected by deformation of the arterial wall. Thus, a coupled model of a blood flow and a deformable arterial wall with filtration flow would be invaluable for simulation of the flow field and concentration field in sequence. However, this type of highly coupled interaction analysis has not yet been attempted. Therefore, we performed a coupled analysis of an artery with multiple bends in sequence. First, based on the theory of porous media, we modeled a deformable arterial wall using a porohyperelastic model (PHEM) that was able to express both the filtration flow and the viscoelastic behavior of the living tissue, and simulated a blood flow field in the arterial lumen, a filtration flow field and a displacement field in the arterial wall using a fluid-structure interaction (FSI) program code by the finite element method (FEM). Next, based on the obtained results, we further simulated LDL transport using a mass transfer analysis code by the FEM. We analyzed the PHEM in comparison with a rigid model. For the blood flow, stagnation was observed downward of the bends. The direction of the filtration flow was only from the lumen to the wall for the rigid model, while filtration flows from both the wall to the lumen and the lumen to the wall were observed for the PHEM. The LDL concentration was high at the lumenwall interface for both the PHEM and rigid model, and reached its maximum value at the stagnation area. For the PHEM, the maximum LDL concentration in the wall in the radial direction was observed at the position of 3% wall thickness from the lumenwall interface, while for the rigid model, it was observed just at the lumenwall interface. In addition, the peak LDL accumulation area of the PHEM moved about according to the pulsatile flow. These results demonstrate that the blood flow, arterial wall deformation, and filtration flow all affect the LDL concentration, and that LDL accumulation is due to stagnation and the presence of filtration flow. Thus, FSI analysis is indispensable.     

Circumventing the heterogeneity and instability of human serum albumin-interferon-alpha2b fusion protein by altering its orientation

Albuferon is a novel long-acting interferon resulted from the direct genetic fusion of human albumin and interferon-alpha2b (HSA-IFN-alpha2b). Albuferon, co-developed by Human Genome Sciences Inc. and Novartis, is currently in late stage development for the treatment of hepatitis C. It was unexpected that HSA-IFN-alpha2b secreted from Pichia pastoris migrated as doublets on non-reducing SDS-PAGE and was prone to form covalent aggregates in aqueous solution. The heterogeneity and instability of HSA-IFN-alpha2b lowered its recovery rate to about 10% and necessitated lyophilized formulation. Site-directed mutagenesis revealed that the heterogeneity and instability of HSA-IFN-alpha2b was caused by the incomplete disulfide bridge formation between Cys1 and Cys98 of IFN-alpha2b. To alleviate the structural perturbation of IFN-alpha2b by HSA, IFN-alpha2b-HSA fusion protein, in which IFN-alpha2b was located at the N-terminus, was created. IFN-alpha2b-HSA was shown to be homogeneous and stable at 37 degrees C for at least 10 days. The improved homogeneity and stability of IFN-alpha2b-HSA increased the recovery rate by 2.5-fold and made the development of stable solution formulation possible. In vitro antiviral assays showed that both fusion proteins retained the activity of IFN-alpha2b, and the EC(50) of HSA-IFN-alpha2b, and IFN-alpha2b-HSA was calculated to be 120+/-12.5, and 160+/-1 1.3ng/ml, respectively. The increased recovery rate and the possibility of solution formulation of IFN-alpha2b-HSA may compensate for its slightly decreased in vitro activity, and makes it to be a promising therapeutic agent that deserves further evaluation.