社区居民对基因检测服务需求意愿及其影响因素调查分析

目的:了解社区居民对基因检测服务的知晓、需求状况及需求意愿的影响因素。方法:采用典型抽样方法,对杭州市下城区1647户居民进行问卷调查。结果:调查人群对基因检测知晓率为21.8%,需求率为12.7%,有需求意愿的占到61.9%。有无家庭成员患病(OR=1.619,95%CI=1.279-2.049)、对基因检测服务知晓情况(OR=2.368,95%CI=1.782-3.146)、有无家族遗传史(OR=2.784,95%CI=1.685-4.600)年龄(x2=27.210,P<0.001)、对婚前基因检测的认可度(x2=23.185,P<0.001)等五个因素对社区居民基因检测服务需求意愿有显著影响。结论:社区居民对基因检测服务需求意愿较强,但是知晓率、需求率均偏低。有待加强相关方面的宣传、教育、培训等工作。     

慢病毒靶向携带Survivin siRNA抗肺腺癌的体内研究

目的:探讨慢病毒载体介导的靶向survivin基因小干涉RNA(survivin-siRNA)对裸鼠移植人肺腺癌的体内抑瘤活性。方法:构建表达survivin-siRNA的慢病毒载体和移植人肺腺癌裸鼠模型,肿瘤组织局部注射survivin-siRNA慢病毒载体,观察肿瘤体积及随时间生长变化;PI染色检测细胞凋亡;流式细胞术检测肿瘤细胞周期变化。结果:慢病毒载体介导survivin-siRNA对裸鼠肺腺癌的抑瘤率为46.07%;30～35%的肿瘤细胞凋亡;G1期细胞比例明显增加,S期细胞比例则明显减少。结论:靶向survivin的RNAi能有效抑制裸鼠移植人肺腺癌的生长,诱导肿瘤细胞凋亡和细胞周期改变。     

硫酸长春碱聚乳酸缓释纳米粒的制剂学研究和体外释放度考察

目的:制备硫酸长春碱聚乳酸纳米粒(VLB-PLA-NPs)并考察其体外释放度.方法:采用复乳挥发法制备VLB-PLA-NPs,以包封率为主要指标评价指标,选择聚乳酸用量、超声时间、外水相浓度为考察因素,优化制备工艺,考察体外释放度.结果:优化工艺制备得VLB-PLA-NPs平均粒径为65±3.03nm,包封率为(99.68±0.30)%,载药量为3.323±0.01μg/mg,体外释放可持续20天.结论:该制备工艺操作简便,结果稳定,缓释特征明显,应用前景良好.     

小剂量α-干扰素联合利巴韦林治疗丙型肝炎肝硬化脾功能亢进的疗效与安全性

目的:观察应用α-干扰素(IFN-α小剂量开始逐渐加量联合利巴韦林治疗丙型肝炎肝硬化代偿期脾功亢进的临床疗效及安全性.方法:14例丙型肝炎肝硬化脾功亢进患者采用IFN-α利巴韦林治疗.IFN-α1MU,隔日一次,皮下注射,两周后对耐受良好的患者,将IFN-α量增加至2MU,以后如能够耐受,则每4周增加1MU,直至5MU,如不能耐受增加剂量,则按原剂量治疗,疗程一年.利巴韦林开始剂量600mg/d,如能耐受逐渐增加至900～1200mg/d,不能耐受按原剂量治疗,疗程一年.观察基因型、血常规、肝功能、HCV-RNA及不良.结果:14例患者,8例为基因1b型;6例为基因2a型.12例患者完成一年疗程.8例出现病毒学应答,应答率66.7%,HCV-RNA平均下降3.6±1.7log10,SVR33.3%,肝功能复常率75%,白细胞、血小板治疗前后比较差异无统计学意义.无不良事件发生.1例合并特发性血小板减少性紫癜(ITP)患者白细胞、血小板恢复正常,PAIg-G明显下降.结论:丙型肝炎肝硬化脾功能亢进患者可选择小剂量IFN-α合利巴韦林逐渐加量治疗方案治疗,未见明显不良反应.     

多巴胺D2受体基因多态性位点与海洛因依赖的关系

目的:探讨多巴胺D2受体(Dopamine D2 receptors,DRD2)基因3'非翻译区Taq ⅠA、启动子区-141 Ins/Del 2个多态性位点和海洛因依赖的相关性.方法:采用聚合酶链反应-限制性片段长度多态(PCR-RFLP)技术检测320例海洛因依赖患者及300例健康对照组的TaqⅠA和-141 Ins/Del2个多态性位点的基因型.采用HaploView4.0及SPSS11.5软件分析这2个多态性位点的基因型、等位基因频率及组间差异.结果:DRD2基因Taq ⅠA位点的基因型及等位基因频率分布在海洛因依赖组与正常对照组存在显著性差异(p<0.01),海洛因依赖组TaqⅠA位点的等位基因A1频率显著高于正常对照组(x2=11.156,p=0.001,OR=1.463,95%CI=1.170～1.830);DRD2基因-141 Ins/Del位点的基因型及等位基因频率分布在海洛因依赖组与正常对照组之间无统计学差异(p<0.05).结论:DRD2基因TaqⅠA位点多态性可能与海洛因依赖有关,携带有TaqⅠA多态性位点A1等位基因的个体可能更容易对海洛因产生依赖.     

甘草酸二铵对于HCV相关性B-NHLCD25-和CD25+T细胞增殖影响研究

目的:本研究初步探究甘草酸二铵(Diammonium Glycyrrihizinate,DG)对HCV相关性B细胞非霍奇金淋巴瘤(B-cellnon-Hodgkin's lymphoma,B-NHL)CD25-T细胞、CD25+T细胞的免疫调控作用。方法:应用流式细胞分析仪检测HCV相关性B-NHL CD4+CD25+T细胞占CD4+T细胞的比例、CD25+细胞占总PBMC比例,并与单纯HCV感染患者、健康人检测结果相对照。应用免疫磁珠分离法(MACS)分选获得CD25-和CD25+细胞,将两者和未分选的外周血单个核细胞(peripheral blood mononuclearcell,PBMC)以CFSE标染孵育72小时后,应用流式细胞分析仪将APC CD3阳性细胞设定为门检测CD25-T细胞、未分选T细胞、CD25+T细胞的增殖情况,及甘草酸二铵干预后的CD25-T细胞、CD25+T细胞增殖情况。结果:应用流式细胞分析仪检测健康人、单纯HCV感染者和HCV相关性B-NHL患者在CD4+CD25+占总CD4+细胞比例和CD25+占总细胞比例上均呈现逐步递增的关系(分别为33.94%±2.18%,57.95%...     

The nucleocytoplasmic transport of viral proteins

Molecules can enter the nucleus by passive diffusion or active transport mechanisms, depending on their size. Small molecules up to size of 50-60 kDa or less than 10 nm in diameter can diffuse passively through the nuclear pore complex (NPC), while most proteins are transported by energy driven transport mechanisms. Active transport of viral proteins is mediated by nuclear localization signals (NLS), which were first identified in Simian Virus 40 large T antigen and had subsequently been identified in a large number of viral proteins. Usually they contain short stretches of lysine or arginine residues. These signals are recognized by the importin super-family (importin α and β) proteins that mediate the transport across the nuclear envelope through Ran-GTP. In contrast, only one class of the leucine-rich nuclear export signal (NES) on viral proteins is known at present. Chromosome region maintenance 1 (CRM1) protein mediates nuclear export of hundreds of viral proteins through the recognition of the leucine-rich NES.     

紫外诱变筛选海洋红酵母S8的尿嘧啶缺陷型菌株

本课题组从海南天然海域筛选到一株高产类胡萝卜素的海洋红酵母菌株S8,该菌株对鱼无毒害,并与鱼共生,欲将其应用于盐诱导表达外源蛋白的海洋红酵母工程菌的构建。本研究利用紫外诱变筛选的方法处理S8菌株,通过统计其UV致死率、5-氟乳清酸致死率等筛选S8的尿嘧啶营养缺陷型突变株。研究结果表明,供试菌株通过紫外线诱变、5-氟乳清酸致死和回复突变率的实验筛选,共获得16株稳定的尿嘧啶缺陷型突变株,突变菌株在基本培养基中培养了8d仍不能生长。选择了其中的一株ST5进行了产胡萝卜素能力的测定,结果表明,在同样的培养条件下,野生型S8菌株细胞生物产量可达87.55g/L,类胡萝卜素含量可达520μg/g,突变株ST5的细胞生物产量为85.45g/L,类胡萝卜素含量为512μg/g;ST5的产胡萝卜素能力方面与野生型S8无明显差异。因此,尿嘧啶缺陷型菌株ST5可为下一步海洋红酵母工程菌的构建提供受体菌。     

Structural plasticity of dendritic spines

Dendritic spines are the major targets of excitatory synaptic input. They exist in a wide variety of shapes and sizes, from thin to mushroom-shaped to stubby. One of the striking characteristics of dendritic spines is their motile nature. Spines can undergo various structural modifications such as changes in density, shape, size, and motility. During development, spines are highly dynamic and many spines are formed and eliminated. As animals mature, most spines become stable and the vast majority of them can last throughout life. However, spine morphology can still undergo progressive changes. Structural dynamics of dendritic spines is thought to play important roles in synapse plasticity and information processing. Abnormal spine structures are often associated with malfunction of the nervous system.     

Nitric oxide synthase activation and oxidative stress,but not intracellular zinc dyshomeostasis,regulate ultraviolet B light-induced apoptosis

AimsTo investigate the role of nitric oxide synthase (NOS) and intracellular free zinc ion (Zn²⁺) in regulation of ultraviolet B light (UVB)-induced cell damage and apoptosis.Main methodsReal-time confocal microscopy measurement was used to determine the changes of intracellular free zinc concentration under different conditions. Cell apoptotic death was determined using fluorescein isothiocyanate (FITC) conjugated-annexin V (ANX5)/PI labeling followed by flow cytometry. Western analysis was used to determine cell apoptosis and eNOS uncoupling.Key findingsUVB induced an elevation of Zn²⁺ within 2 min of exposure. The UVB-induced intracellular Zn²⁺ elevation was dependent on the increase of constitutive nitric oxide synthase (cNOS) activity and production of superoxide. Removal of Zn²⁺ with a lower concentration (< 25 μM) of N,N,N′,N′-tetrakis (2-pyridylmethyl) ethylenediamine (TPEN), a Zn²⁺-specific chelator, did not induce cell death or prevent cells from UVB-induced apoptosis. However, a higher [TPEN] (> 50 μM) was cytotoxic to cells, but prevented cells from further UVB-induced apoptosis. The higher [TPEN] also induced cNOS uncoupling. Furthermore, treating the cells with a membrane permeable superoxide dismutase (PEG-SOD) inhibited Zn²⁺ release and reduced apoptotic cell death after UVB treatment. The results demonstrated a complex and dynamic regulation of UVB-induced cell damage.SignificanceOur findings not only advance our understanding of the correlations between cNOS activation and Zn elevation, but also elucidated the role of cNOS in regulation of oxidative stress and apoptosis upon UVB-irradiation.