柴郁地仙方治疗围绝经期抑郁症体会

围绝经期抑郁症的发病基础是肾虚精血亏少,七情过极是诱因,病程中多表现为肝郁气滞、痰瘀交阻、闭塞脑窍。肾虚肝郁又常累及心脾二脏,故以补肾疏肝、化痰祛瘀、补益心脾立法,自拟柴郁地仙方治疗本病取得良好效果。从实验及临床研究结果来看,补肾疏肝、化痰祛瘀、补益心脾这种多脏腑整体调治,虚实兼顾治疗围绝经期抑郁症的思路较有效,且值得进一步推广和深入探讨。福建省卫生厅资助项目(WZPW101307);; 厦门市科技局资助项目(3502Z20133007

Preparation and determination of polyclonal antibody to thymosin α1

目的 :制备用于检测转胸腺素基因蓝藻表达产物的特异性抗体。方法 :用提纯的小肽 (2 8个氨基酸 )胸腺素α1与牛血清白蛋白偶联后作为抗原 ,采用皮下多点注射免疫的方法免疫大鼠。经过 3个月的免疫 ,获得抗Tα的多克隆抗体。结果 :用间接酶联免疫吸附 (ELISA)方法 ,测得抗体效价超过 40 96。蛋白印迹 (Westernblot)检测结果显示 ,该抗体能特异性地与胸腺素α1抗原产生明显免疫亲和反应。结论 :所制备的抗体具有很好的灵敏性和特异Objective:To prepare specific antibody against Thymosin α1 Methods:Thymosin α1(Tα1,28 peptide) was conjuncted with BSA as immune antigen,rats were immunized and the polycolonal antibody to Tα1 was obtained.Results:With ELISA detection,the titer of antibody was more than 1:4 096;Western blot analysis showed that the antibody can bind with Tα1 specifically.Conclusion:The prepared antibody against Tα1 has good specificity and reactivity and can be used to detect the expression products of transgenic cyanobacteria which expressed Tα1 gene.国家“863”课题资助!(No 819 0 4 0 3

重庆市主城区长江和嘉陵江水中有机污染物对斑马鱼胚胎仔鱼的毒性研究

目的以重庆市主城区大溪沟(嘉陵江)和寸滩(长江)两个点为代表,研究2004~2005年度重庆市水源水中有机污染物的组成和对斑马鱼胚胎仔鱼的毒性。方法固相萃取法萃取水中有机污染物,GC/MS检测有机污染物的种类;将萃取的有机污染物溶于DMSO中,并用斑马鱼胚胎仔鱼实验研究水中有机污染物对斑马鱼胚胎孵化率和仔鱼畸形率的影响。结果四个水样中均可检出有机污染物,污染物的种类以酞酸酯类和多环芳烃类为主;污染物可导致斑马鱼胚胎孵化率降低及仔鱼畸形率增加,其毒性呈现出时间和剂量依赖性;同一采样点中,枯水期水样毒性大于

CONSTRUCTION OF SHUTTLE EXPRESSION VECTOR AND EXPRESSION OF THYMOSIN α_1 IN SYNECHOCOCCUS SP.PCC7942

利用本实验室构建的含蓝藻Plectonema boryanum内源小质粒的穿梭质粒pPRS-1,改建成含热诱导启动子、泛素融合的胸腺素α_1(UB-Tα_1)目的基因、卡那霉素抗性选择标记、rbcs终止子的新的穿梭表达重组质粒pPRFUT。将这种重组质粒转化蓝藻Synechococcus sp.PCC7942,通过抗性筛选获得了具卡那霉素抗性的转化藻株。经Southern-blot杂交证实,穿梭表达质粒已转入蓝藻Synechococcus sp. PCC7942细胞中;在42℃热诱导30min后,目的基因UB-Tα_1得到较高水平表达,表达量约占总蛋白量的7.5%。The shutter expression vector pPREUT was constructed from the plasmid pPRS-1 containing the en dogenous snail plasmid of Plectonema boryanum. The expression elements such as heat shock gene groESL promoter, foreign gene Ub-thymosin α1, rbcS polyA terminator and Kanamycin resistance gene were all included. The shutter plasmid pPREUT was direcdy transferred into Synechococcus sp. PCC7942. The transformants were obtained through Kanamycin screening. Southern blotting analysis showed that the shutter plasmid have been transferred into Synechococcus sp. PCC7942. After induction by heat shock(42℃) for 30 min, the foreign protein LIR-Tα1 was ex pressed efficiently, which reacted 7.5% of total amount protein.国家863资助项目(863-819-04-03

Construction of High Level Expression System by Heat Shock Induced in Cyanobacterium

以聚球藻Synechococcussp .PCC794 2作为基因工程受体菌 ,构建了热休克诱导的高效表达系统。用PCR法扩增并克隆出Synechococcussp .PCC794 2自身的热休克基因 groESL操纵子的强启动区 (2 4 0bp) ,并紧靠其后组装上泛素 (UB)融合的胸腺素α1(Tα1)目的基因 ,在下游组装有rbcSpolyA终止子。此外 ,还组装了一个完整的卡那霉素抗性基因作为筛选标记基因。这样 ,就构建了完整的蓝藻的基因表达系统。经转化蓝藻Synechococcussp .PCC794 2细胞 ,在 4 2℃热诱导 30min后 ,目的基因UB Tα1得到较高水平表达 ,表达量约占可溶性蛋白的 8.7%A high level expression system by heat shock induced had been constructed for Synechococcus sp. PCC7942. First, the strong promoter (240bp) of heat shock gene groESL operon from Synechococcus sp. PCC7942 was cloned by PCR. Then the ubiquitin fused thymosinα 1 gene(UB Tα 1)?rbcS polyA terminator were subcloned sequentially in the downstream of groESL promoter. The kanamycin resistance gene, which as the marker gene, was also subcloned into this expression system to obtain the expression vector pEUTMT 1. Induced by 42℃ heat shock for 30 min, the level of UB Tα 1 protein reached a value of 8.7% of the total soluble protein in the transformants of Synechococcus sp. PCC7942. The result of the experiment demonstrates that the heat shock induced high level expression system of Synechococcus sp. PCC7942 was constructed successfully.863计划资助项目 (863 819 0 4 0 3