微透析法获取鹅掌柴嫩茎质外体汁液的研究

该文研究的重点是获取质外体汁液。假设微透析技术可以在植物上得到质外体汁液,并以盆栽鹅掌柴为材料,在活体嫩茎插入微透析探针后的不同时间段内分别收集透析液,测定其中的离子浓度和苹果酸脱氢酶活性加以验证。结果表明:在探针插入120 min后,收集的透析液中Na+、K+和Ca2+浓度逐渐趋于平缓,并且苹果酸脱氢酶的活性消失,从而证明120 min后得到的微透析液,没有受到破损细胞液的污染,是纯净的鹅掌柴嫩茎质外体汁液，从而为活体、方便、快捷地获取植物质外体汁液提供新的方法。      

含LacZ重组逆转录病毒的制备及其在NIH3T3细胞的表达

构建了含有3．7kbLacZ的重组逆转录病毒载体,转染包装细胞系PA317后,经G418筛选,得到了G418的抗性的产毒细胞系,用收获的含有重组逆转录病毒粒子的浓缩病毒上清,感染NIH3T3细胞。经X-gal染色检测,发现表达了LacZ的NIH3T3细胞呈蓝色。     

稻鸭共生防虫增收操作技术

稻鸭共生具有除草、除虫、培肥、中耕、促进稻株发育等特点。生产上综合效益显著,其产品为绿色无公害有机食品。稻鸭共生是种养集成技术,它构成农业自然生态体系,是农民增产、增收的途径,是农业持续发展的需要,值得大力推广。宿松县千岭地区2002～2004年在龙湖圩田、金川垄田、坂田、榜田大面积试验示范,取得了显著效益。1稻鸭共生的优良效果1.1除草经长期田间观察,鸭子喜食禾本科以外的植物和水面浮游杂草,有时也吃幼嫩的禾本科植物。鸭子在稻田活动,一刻也不停息的嘴和脚具有较好的除草作用,能干净地采食践踏牛毛草、鸭舌草、四叶草、节节…     

茉莉酸诱导小麦抗病虫性初步研究

初步研究了茉莉酸诱导对小麦苗抗病虫能力的影响,结果显示,小麦在喷施茉莉酸后能够提高植株对麦长管蚜和小麦白粉病菌、小麦叶锈病菌的抵抗能力,可显著降低小麦白粉病、叶锈病的发病级别和病斑数量,对麦长管蚜则在体重和产仔数量上有显著的抑制作用。     

螺旋毛壳ND35 β-1,3-葡聚糖酶的诱导、性质及其抑菌作用

 以病原菌Rhizoctonia solani的细胞壁为诱导物,模拟毛壳菌自然的重寄生过程,研究了内生真菌螺旋毛壳(Chaetomium spirale) ND35 β-1,3-葡聚糖酶的产酶条件、性质,尤其是不同碳源的调控作用。结果表明,不同种类的真菌细胞壁及几丁质和昆布多糖,均可诱导产生β-1,3-葡聚糖酶,而作为分解代谢产物的葡萄糖则抑制产酶。经硫酸铵沉淀、DEAE-Sepharose阴离子交换层析及Phenyl-Sepharose疏水层析,并通过SDS-PAGE鉴定,纯化了一种分子量约为73 kDa的内切β-1,3-葡聚糖酶GLUC73。其最适反应温度为55℃,在40℃以下较稳定;最适pH值为5.5,在pH 5-9范围内均很稳定;酶活性受Hg²⁺、Fe³⁺、Zn²⁺、Mg²⁺等金属离子不同程度的抑制,Mn²⁺和Co²⁺对酶有激活作用;以昆布多糖为底物时,该酶的米氏常数K_m为0.412 mg·mL^-1,最大反直速度V_max为3.876 U·mL^-1。粗酶液同时具有β-1,3-葡聚糖酶和几丁质酶活性,离体抑菌试验表明,对苹果炭疽病菌(Glomerella cingulata)、杨树腐烂病菌(Valsa sordida)、苹果树腐烂病菌(Valsa mali)的菌丝生长和孢子萌发有明显的抑制作用。通过对β-1,3-葡聚糖进行免疫细胞化学标记和超微结构观察,间接证明了β-1,3-葡聚糖酶在螺旋毛壳重寄生过程中的作用。     

RT-PCR结合限制性内切酶分析法区分NDV强弱毒株

通过设计的特异引物，采用RT-PCR对1996年以来分离的45株国内新城疫病毒分离株进行鉴定和分析，并与其MDT实验结果进行比较，建立了RT-PCR结合BglⅠ内切酶分析区分新城疫强弱毒的方法。此方法操作简便、迅速，不仅能区分新城疫的强弱毒株，还可诊断AMPV-1引起的其他禽类的感染。     

Overexpression of neuronal ARNT2 is involved in neuronal apoptosis evoked by low K+

AIM: To observe the expression of neuronal Aryl hydrocarbon receptor nuclear translocator 2 (ARNT2) involved in neuronal apoptosis evoked by low K+ and to investigate the relationship between ARNT2 and neuronal apoptosis. METHODS: After neuron apoptosis model was established, the changes of mRNA and protein of ARNT2 during apoptosis were investigated by RT-PCR and Western blotting, respectively. Immunofluorescence was analyzed by confocal microscopy to probe the subcellular localization of ARNT2. RESULTS: Induced by low K+, the expression of ARNT2 mRNA was up-regulated obviously at the point of 30 min, and peaks at the point of 1 h. This up-regulated expression lasted for 12 h, and the variation of ARNT2 protein was similar to that of mRNA. The results of immunofluorescence analyzed by confocal microscopy showed that the localization of ARNT2 protein was in the nucleus. CONCLUSION: ARNT2 locate in nuclei of normal cerebellar granule neurons of rat. During the process of apoptosis evoked by low K+, both mRNA and protein of ARNT2 are overexpressed.     

Hypoxia upregulates CD73 expression in mouse microvascular endothelial cells

AIM: To study the effect of hypoxia on CD73 expression in mouse microvascular endothelial cell line bEnd.3. METHODS: ① bEnd.3 cells were exposed to different periods of hypoxia. ② Concentration of LDH released by bEnd.3 cells into the culture medium was detected. ③ Surface CD73 activity in bEnd.3 cells was measured by HPLC according to the conversion of E-AMP to E-ADO. ④ CD73 mRNA expression were analyzed by semiquantitative RT-PCR. ⑤ Cell surface proteins were biotinylated and CD73 was detected by avidin blots of immunoprecipitation with mAb TY23. RESULTS: ① bEnd.3 cells exposed to hypoxia for 24 h demonstrated a significant increase in LDH release (P<0.01). ②Hypoxia induced CD73 activity in bEnd.3 cells in a time-dependent manner. ③ CD73 mRNA expression increased markedly in hypoxia for 4 h and 8 h (P<0.05). ④ bEnd.3 cells exposured to hypoxia induced a time-dependent increase in expression of CD73 (P<0.05). CONCLUSION: Hypoxia induces CD73 mRNA, protein expression and increases CD73 activity in mouse microvascular endothelial cells.     

Effects of antibiotic treatment on the inflammatory mediator expression in peritoneum and the peritoneal transport function in the rat with acute bacterial peritonitis

AIM: To observe the effect of antibiotic treatment on the inflammatory mediator expression in peritoneum and the peritoneal transport function in rats with acute peritonitis, and explore its mechanisms. METHODS: Eighty-six SD rats were randomly divided into three groups. Control group (n=28) were treated with PBS (ip), peritonitis group (n=28) and treatment group (n=28) were challenged with the E.coli (ip), but at 3 h and 9 h gentamicin was given (ip) in treatment group. Seven rats of every group were randomly sacrificed at 24 h, 48 h, 72 h and 7 d. Peritoneal equilibration test (PET) was did before they were killed. Leukocyte count, pathological changes and the expression of CD45, NF-κB， IL-1β， TNF-α in peritoneum were examined. RESULTS: (1)The blood leukocytes in peritonitis group decreased strikingly, but did not change obviously in other two groups. The peritoneal fluid leukocytes in peritonitis group increased significantly from 24 h to 72 h, while in treatment group, it enhanced more strikingly than peritonitis group at 24 h, and recovered earlier. (2) Both in peritonitis group and treatment group, the expression of activated NF-κB, IL-1β, TNF-α and CD45 increased significantly, but the treatment group was lower than model group at 48 h and 72 h. The mRNA level of IL-1β and TNF-α had the same trend as their protein expression. (3) Compared with the control group, UF and D/D0 Glu decreased significantly in model group and treatment group, and D/PTP increased dramatically. The D/P TP in treatment group lowered obviously compared with peritonitis group, while the net UF and D/D0 Glu had not significant difference between treatment group and model group. CONCLUSION: Antibiotic treatment can partly decrease the expression of inflammatory mediators in peritoneum of rats with acute peritonitis and also can improve the protein transport ability to some extent, but can not improve the peritoneal ultrafiltration and the glucose transport function.     

Effects of ibrolipim against atherosclerosis in minipigs

AIM: To explore the application mechanism of NO-1886 (ibrolipim), a synthetic compound, improving dyslipidemia and inhibiting atherosclerosis in Guizhou minipigs fed with high fat/high sucrose diet. METHODS: Fifteen Chinese Guizhou minipigs were randomized into three groups with similar body weight ［(n=5 in normal control group (CD); n=5 in high fat/high sucrose group (HFSD); n=5 in high fat/high sucrose supplemented ibrolipim group （HFSD+ibrolipim)］. Blood samples were withdrawn from the eyehole sinus venosus of the animals at the end of each month after fasting overnight. The animals were sacrificed at the end of 8 months. The concentrations of cholesterol ester in plasma HDL were analyzed by HPLC. The aortic fatty streak-lesions were quantified following lipid staining with Sudan IV. Lipid droplets in liver were observed by Oil red O staining. RESULTS: Compared with CD, fasting plasma TC, TG and FFA levels of HFSD were elevated significantly. The aortic fatty streak-lesions were clearly presented in the animals’ aortas. The intima became rougher and thicker. A lot of lipoid foam cells migrated to regions of intima and smooth muscle cells, which associated with the injuries of internal elastic lamina. Extensive fat deposited in the liver were observed. Supplementing of 1.0% ibrolipim into high fat/high sucrose diet induced the decrease in plasma TG and FFA concentrations and an increase in plasma HDL-C concentration compared with HFSD. A little fat deposited in the liver were observed. CONCLUSION: ibrolipim prevents AS in high fat/high sucrose diet feeding minipigs through decreasing the plasma TG and elevating the plasma HDL-C.