苹果柱型基因Co的一个AFLP 标记的SCAR转换

 将苹果柱型基因的一个AFLP 标记成功地转换成了简单实用的SCAR 标记。首先对AFLP 标记片段进行序列测定, 然后根据序列特点设计了两对特异引物CoA1/ CoA2 和CoA1/ CoA3 , 每条引物长20 bp。PCR 结果表明CoA1/ CoA2 可以扩增出216 bp 和148 bp 两条带, 其中216 bp 的为柱型性状的特征带; CoA1/CoA3 可以扩增出273 bp 和205 bp 的两条带, 其中273 bp 的为柱型性状的特征带。两对引物在杂交后代中扩增出的特征带与柱型性状的分离重组率都很低(CoA1/ CoA2 为6. 3 % ±2. 5 %; CoA1/ CoA3 为7. 3 % ±2. 6 %) , 所以它们都可以作为该SCAR 标记的特异引物所用。     

High efficient expression of human endostatin in E.coli and its antiangiogensis activity

AIM: To examine the expression of human endostatin in E.coli, produce its fusion protein antibody and observe its biological activity. METHODS: Endostatin gene was amplified by polymerase chain reaction,recombined with plasmid vector pGEX-2T and induced expression with IPTG.The protein activity was tested by endothelial cell proliferation inhibitory assay.Inclusion body crudely purified was used to generate polyclonal antibody to detect its expression at mouse's liver and kidney etc. RESULTS: The protein expressed was 20kD after digestion by thrombin,it appeared the anti-angiogenesis activity and Western blotting indicated the expression of endostatin in liver and kidney of mouse. CONCLUSION: The successful expression of human endostatin and the preparation of polycolonal antibody indicated its potential application in anti-angiogenesis therapy and diagnosis tumors.     

典型光活化毒素α-三噻吩对棉铃虫和亚洲玉米螟Na~+-K~+-ATPase的抑制作用

以棉铃虫 H elicoverpa armigera和亚洲玉米螟 Ostrinia furnacalis为试虫 ,建立 Na+- K+-ATPase最佳反应系统 ,研究典型光活化毒素 α-三噻吩 (简称 α- T)对 Na+- K+- ATPase的影响。棉铃虫 Na+- K+- ATPase活力最佳测试条件为酶源蛋白浓度 6μg/ m L ,反应温度 35～ 4 0℃ ,反应时间6min;亚洲玉米螟则为酶源蛋白浓度 8μg/ m L,反应温度 35℃ ,反应时间 6min。近紫外光照 (30 0～4 0 0 nm )对棉铃虫和亚洲玉米螟离体 Na+- K+- ATPase活力基本没有影响 ,但对活体活力有很强的抑制作用。光照和无光照条件下 ,α- T对两种昆虫离体和活体 Na+- K+- ATPase活力均有不同程度的抑制作用。光照组 α- T对亚洲玉米螟 Na+- K+- ATPase的抑制率高于无光照组 ,且处理浓度或剂量越高 ,其抑制率越大 ;对棉铃虫 Na+- K+- ATPase抑制作用不显著     

钻蛀板栗枝干害虫的发生情况分析

本文对钻蛀板栗枝干害虫的发生分布、致树木死亡的发生量和不同树龄期的发生差异等情况进行了分析。从中找到了一些规律，可供有关方面应用。     

磷化氢熏蒸处理对嗜卷书虱不同虫态的致死作用

实验室条件下系统研究了磷化氢(PH3)对储物害虫嗜卷书虱Liposcelis bostrychophila卵、各龄若虫和成虫的致死作用,并选用PH3间歇熏蒸以及PH3与气调交替处理等措施对嗜卷书虱进行处理,比较了不同处理措施对嗜卷书虱种群的控制效果。结果表明,PH3熏蒸处理对嗜卷书虱各虫态有不同的致死效果。对卵而言,24、72和120 h熏蒸处理的LC50分别为0.137、0.045和0.035 mg/L;而24 h熏蒸处理对若虫的LC50在4.285~7.364 μg/L之间,对成虫的LC50为20.404 μg/L;采用25 μg/L的PH3进行24 h熏蒸处理,间隔10 d后再分别进行第2次和第3次熏蒸处理,可以完全控制嗜卷书虱的发生。采用PH3 (12 μg/L)和气调(体积比例为35% CO2,1% O2,64% N2)交替处理能够延缓嗜卷书虱种群抗性的发展,交替处理3~5次可以完全控制嗜卷书虱的发生。     

应用电导仪测定番茄灰霉病菌对多菌灵抗药性的初步研究

1.0 mg/L和5.0 mg/L多菌灵盐酸盐溶液处理灰霉病菌多菌灵敏感菌株和抗性菌丝体,用电导仪测定溶液电导率的动态变化,结果表明:在敏感菌株存在时,多菌灵盐酸盐溶液2 h后的电导率显著降低,而抗性菌株存在的溶液中电导率则未见下降。     

江河源区草地资源现状及其可持续发展

在分析青海省江河源区草地资源现状的基础上，进一步剖析了造成该区域草地生态环境恶化的综合因素，提出了合理利用和保护天然草地，优化草地生产结构，加强基础设施建设，建立饲草料生产基地，遏制草地退化，使草地生态进入良性演替，实现江河源区草地资源可持续利用和草地畜牧业可持续发展的对策。     

Antitumor effect of chlorophyllin in vitro

AIM: To study the effect and mechanism of chlorophyllin (CHL) inhibiting HT29 cells. METHODS: IC₅₀ value and growth curve of HT29 cells were detected with MTT method. Apoptosis was detected with Wright-Giemsa staining, FCM and DNA electrophoresis. Telomerase was detected by PCR-ELISA, and protein and mRNA expression of COX-2 gene were detected through RT-PCR and Western blot. RESULTS: CHL inhibited the growth of HT29 in a dose-dependent manner. CHL blocked HT29 cells in G₁ phase but did not induce apoptosis. Different concentration of CHL inhibits the expression of telomerase and COX-2 in HT29 cells. CONCLUSION: CHL inhibited the growth of HT29 cells by inhibiting the expression of telomerase and COX-2 and blocking cells in G₁ phase.     

Expression of cyclooxygenase-2 in IL-1β-stimulated mesangial cells is medicated by NF-κB/IκB signal pathway

AIM: To investigate the role of NF-κB/IκB signal pathway in the regulation of cyclooxygenase-2 (COX-2) expression in human mesangial cells (HMC). METHODS: The PGE₂ concentration in supernatants of HMC was measured by radioimmunoassay. COX-2 mRNA and protein expression were determined by RT-PCR and Western blot. Electrophoretic mobility shift assay (EMSA) and Western blot were used to detect the activity of NF-κB and degradation of IκB. RESULTS: IL-1β significantly upregulated COX-2 expression and PGE₂ production in HMC. Significant up-regulation of NF-κB activation, nuclear translocation of p65 subunit, and degradation of IκB α and IκB β were observed in IL-1β-induced HMC. CONCLUSION: Expression of COX-2 in IL-1β-induced HMC is mediated by NF-κB/IκB signal pathway.     

Inhibition of K562 cell growth by antisense drug targeting with VEGF mRNA in vitro

AIM: To investigate inhibition of K562 cell growth by antisense drug targeted VEGF mRNA. METHODS: X7, 20-mer antisense sequences were selected, synthesized and modified with phosphorothioate. The drug was transfected into K562 cells in the present of lipofection. Cell growth was assayed by trypan blue dye exclusion assay and MTT. The level of VEGF protein in the media was determined by ELISA. The morphology of apoptotic cells were observed by Giemsa staining, and the propotion of apoptotic cells was detected by flow cytometry. RESULTS: The antisense drug inhibited growth of K562 and downregulated expression of VEGF protein significantly, compared with Scrambed control group and showed dose-dependent relation. Signs of apoptosis of K562 cells were not observed. CONCLUSION: Inhibition of K562 cell proliferation, but not cells apoptosis induction is the mechanism of inhibing growth of K562 cells by antisense drug targeted VEGF mRNA. At same time, VEGF has function of promoting K562 cell proliferation, and VEGF mRNA may be a new target attached by drugs.