新型取代氨基(或芳氧)磺酰基苯氧丙酸酯的合成及其除草活性

由α-苯氧基丙酸酯出发,合成了取代氨基(或芳氧)磺酰基苯氧丙酸酯,并测定了它们的除草活性。所有的化合物均经1H NMR和元素分析确证,初步的生测结果表明上述化合物具有一定的除草活性。     

双孢蘑菇性亲和性相关分子标记的初步筛选

以传统的形态，生理生化分析和最新的DCS－PDMA性亲和性测定方法为基础， 结合群体分离分析和RAPD技术来源于同一双孢蘑菇异核体菌株的12个不育同核原生质体个体进行分析，筛选与性亲和性相关的分子标记。研究结果表明，供试的12个不育同核原生质体个体被分成两大类性亲和性类型，其中一类（A^ ）包括不育同核原生质体个体B、C、D、E、F、G、H、I、J、L，另一类（A^-）则仅仅包括不育同核原生质体个体K和M，同时筛选到一个与性亲和性相关的分子标记OPA16 1500。从而为间地利用双孢蘑菇本身特有的交配型作标记来指导杂交育种工作和进一步将性亲和性基因定位分离克隆奠定了坚实的基础。     

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。     

Study on integrin-mediated albumin microbubbles adherence to activated leukocytes

AIM:To investigate the mechanism responsible for albumin microbubbles adherence to activated leukocytes. METHODS: In vitro studies were performed in which activated or nonactivated leukocytes were incubated with albumin microbubbles and observed under microscopy. The suspensions of leukocytes and microbubbles which contained or absented of integrins were analyzed with flow cytometry.RESULTS: A minimum of 50cells were identified under transillumination. 5 min after microbubbles were incubated with leukocytes, the number of cells interacting with microbubbles was greater for activated cells than for nonactivated cells(20.30±2.67 vs 4.50±1.43, P <0.01).Microbubbles attached to the surface of activated leukocytes were phagocytosed and remained intact for up to 30min. Microbubble attachment was inhibited notably by blocking the leukocyte β₂-integrin Mac-1(P <0.01) and by VLA-4mAb slightly(P <0.05) CONCLUSION: The mechanism of albumin microbubbles attaching to and phagocytosed by leukocytes was due to β₂-integrin and VLA-4 mediation. Phagocytosed microbubbles can remain at the regions of inflammation for15 min, also responsible to ultrasound.     

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     

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.     

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.     

Regulation of MMP-2 and TIMP-3 expression by uPA signal transduction system in human bone giant cell tumor

AIM: To study effects of urokinase-type plasminogen activator (uPA) signal transduction on expression of matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of matrix metalloproteinase-3 (TIMP-3) in giant cell tumor of bone (GCT). METHODS: Expression of uPAR, MMP-2 and TIMP-3 in GCT tissue was detected by immunohistochemistry. Phosphorylation level of mitogen-activated protein kinase (p44) in uPA/uPAR signal pathway in cultured GCT cells was detected by immunoprecipitation. The expression of MMP-2 and TIMP-3 in cultured cells after treatment with uPA-ATF or anti-uPAR antibody was also detected by Western blotting. RESULTS: 1) Urokinase-type plasminogen activator receptor (uPAR) was positive on the cell membrane and in cytoplasm of some mononuclear stromal cells (MSCs) and multinucleated giant cells (MGCs); 2) MMP-2 was positive in the cytoplasm and on the cell membrane of almost all of MSCs and some of MGCs. The polar distribution of MMP-2 in the cytoplasm of MGCs was especially obvious; 3) The expression of TIMP-3 of some MSCs and MGCs in GCT was much lower than MMP-2. The positive signal also showed a prominent polarity; 4) After treatment with uPA-ATF, the phosphorylation level of p44 in GCT cultured cells was much higher than the control. Addition of anti-uPAR antibody in the cells remarkably down-regulated the phosphorylation level of p44 as compared with the control group, suggesting that uPA-ATF participates cell signal transduction and this reaction can be inhibited by anti-uPAR antibody; 5) uPA-ATF cell signal pathway up-regulated expression of MMP-2 and TIMP-3, while anti-uPAR antibody down-regulated the expression of MMP-2 and TIMP-3. CONCLUSION: These results demonstrate for the first time that uPA-ATF directly regulates the expression of MMP-2 and TIMP-3 by signal transduction pathway, and the over-expression of MMP-2 and TIMP-3 may play an important role in local osteolysis of GCT.     

Expression of DNA repair gene ERCC1 and its relationship with PAH-DNA adducts in lung cancer tissues

AIM: To investigate the expression of nucleotide excision repair gene ERCC1 and its relationship with PAH (polycyclic aromatic hydrocarbons)-DNA adducts in lung cancer tissues. METHODS: ERCC1 mRNA expression and the PAH-induced DNA adducts were detected in 150 lung cancer tissues, 120 adjacent lung tissues without cancer cells, 40 benign lung lesions and 40 normal lung tissues. The effects of some exposure factors on the expression of ERCC1 gene and the connection between ERCC1 and PAH-DNA adduct was analyzed. RESULTS: Reduced expression levels of ERCC1 were observed in 46 of 150 (30.7%) lung cancer specimens and 1 of 40 (2.5%) normal lung tissues. Smoking may suppress the expression of ERCC1 gene. The level of PAH-DNA adduct was negatively correlated with the expression of ERCC1 gene, the Spearman coefficient was -0.648, P<0.01. CONCLUSION: ERCC1 is an important nucleotide excision repair gene and may participate in the repair of DNA damage, such as PAH-DNA adduct. Low expression of ERCC1 may play an important role in the development of human lung cancer.     

维生素E对绵羊鲜精及冻精精液品质的影响

日粮中添加维生素E可以提高绵羊鲜精的活率 ,对照组和试验组的活率分别为 0 72± 0 0 7和 0 78± 0 0 6(P <0 0 5 ) ,显著改善鲜精精液品质。采用两步稀释法在绵羊冷冻精液稀释液中添加维生素E ,可以极显著降低冷冻对精子顶体的冷刺激损害程度 ,试验组的活率 ( 0 43 5± 0 0 2 0 )极显著高于对照组 ( 0 3 65± 0 0 2 6) (P <0 0 1) ,精子顶体的总异常率试验组 ( 3 3 72 % )极显著低于对照组 ( 4 4 3 5 % ) (P <0 0 1) ,其中试验组顶体膨胀率和脱落率与对照组分别为 1 2 8%±0 5 8%、2 3 0 8%± 1 45 %与 2 68%± 0 5 9%、2 8 96%± 3 14 %。冷冻精液品质显著改善。