转录靶向猪瘟病毒3''''-UTR shRNA细胞株的初步建立

利用质粒载体在细胞内转录并加工成siRNA的方法，设计3对针对猪瘟病毒3＇-UTR不同位点的干扰片段，分别与干扰载体pGenesil-1连接，转化DH5a，筛选阳性克隆得到重组干扰质粒pGene-1，pGene-2和pGene-3，脂质体介导转染重组干扰质粒于PK-15细胞，G418抗性筛选得到转录靶向猪瘟病毒3＇-UTRshRNA的PK-15细胞株，为今后应用RNAi研究猪瘟病毒Y-UTR调控病毒复制的功能以及抑制猪瘟病毒增殖的研究奠定了基础。     

6HW-50型车载高射程喷雾机防治杨潜叶跳象试验

报道了使用6HW-50车载高射程喷雾机,采用1.2%苦烟乳油不同施药量进行杨潜叶跳象防治的对比试验的结果,旨在评估这一新药械防治杨潜叶跳象的效果.     

Alpha-amylase treatment increases extractable phenolics and antioxidant capacity of oat (Avena nuda L.) flour

In this work, α-amylase was used to treat oat flour with the intent to release phenolic compounds with known antioxidant properties. After methanol extraction, the amounts of nine beneficial phenolic compounds were measured using HPLC. The antioxidant activities of the extracts were assessed using 2,2′-azinobis (3- ethylbenzothiazoline-6-sulfonic acid) (ABTS)，2,2-diphenyl-1-picrylhydrazyl (DPPH), ferric reducing antioxidant power (FRAP) and protein oxidative damage protection assays. Compared with heating-only treated oat flour, that treated with α-amylase showed significant increase of extractable total phenolic content (0.46–1.35 μmol gallic acid equivalents per gram oat), total antioxidant capacity, and an increased ability on the protection of protein from oxidative damage. Heating-only increased caffeic acid and vanillin content by 17 (0.03 vs 0.54 μg/g oat) and 1.8 (0.62 vs 1.11 μg/g oat) folds, but slightly increased the content of other phenols. Excluding heating effect, α-amylase treatment increased gallic acid content by 2.6 folds (0.38 vs 1.38 μg/g oat), caffeic acid content by 2.4 (0.54 vs 1.82 μg/g oat) folds, and other phenols by 1.0–1.8 folds. In conclusion, α-amylase treatment can yield oat products containing more extractable phenolic compounds with increased antioxidant capacity.     

Micropropagation of dahlia in static liquid medium using slow-release tools of medium ingredients

Growth of dahlia shoots in vitro was ca. 4 times faster in liquid medium than on solidified medium. In liquid standard medium (3% sucrose, macroelements according to Driver–Kuniyuki Walnut medium, microelements according to Murashige–Skoog medium, 0.44 μM benzylaminopurine), the major medium ingredients were consumed for 75–80% during the first 6 weeks. Addition of extra ingredients increased growth, demonstrating that the amount of ingredients added at the start of culture was suboptimal. When the extra ingredients were given at the start of the culture, concentrations became too high and therefore inhibitory. When the ingredients were added during the subculture cycle by means of small aliquots of a concentrated solution or by means of slow-release tools, growth was strongly increased. Osmocote gave satisfactory results as a slow-release tool for inorganics. For organic ingredients (sucrose and benzylaminopurine), a novel slow-release tool was developed.     

两系杂交稻扬两优6号的选育及栽培特性研究

扬两优6号系江苏里下河地区农业科学研究所用广占63—4S与扬稻6号杂交配组育成的两系杂交籼稻新组合。该组合集优质、高产、多抗、适应性广于一体,先后通过江苏、贵州、河南、湖北等省和国家审定定名,已在南方稻区10多个省(市、区)推广应用,是发展粮食生产、促进品种结构调优、推动优质杂交水稻种子和优质稻米产业化发展的主体品种。     

Ag-ELISA and PCR for Monitoring the Vaccination of Cattle against Taenia saginata Cysticercosis Using an Oncospheral Adhesion Protein (HP6) with Surface and Secreted Localization

A Taenia saginata oncosphere-derived adhesion protein (HP6) with surface and secreted localization was used to successfully vaccinate calves against oral challenge with T. saginata eggs. In contrast, vaccination using a combination of T. saginata oncosphere-derived peptides, selected on the basis of their antigenic index, and including three derived from the HP6 molecule (HP6-1, HP6-2 and HP6-3), was unsuccessful. This either indicated that the wrong peptides were selected or, in the case of the HP6 protein, that the protective epitope is conformational in nature. The protection experiments were monitored using a parasite antigen detection ELISA (HP10 Ag-ELISA), which allowed the early determination of the success of the vaccination protocol, subsequently confirmed at autopsy. PCR assays were used for the first time to confirm the presence of T. saginata DNA in lesions recovered at autopsy and thus verify the parasite origin of the lesions.     

用两种方法生产猪丹毒(G4T10)活疫苗的比较试验

为提高猪丹毒(G4T10)活疫苗培养菌数,采用加NaOH调pH增菌和加吐温-80增菌两种方法进行比较试验,前一种方法生产152批,平均培养菌数为7.7×109 CFU/mL.后一种方法生产50批,平均菌数为1.792×1010 CFU/mL.结果表明,后一种方法较为理想.     

Effects of NS-398 on the proliferation and apoptosis of HepG2 cells

AIM: To evaluate the effects of NS-398, a cyclooxygenase-2(COX-2) inhibitor, on the proliferation and apoptosis in HepG2 cells. METHODS: The effects of NS-398 on the proliferation of HepG2 cells was evaluated by MTT. DNA fragmentation gel analysis was used to analyze the apoptotic cells; DNA ploidy and apoptotic cell percentage were examined by flow cytometry. Furthermore, the expression of COX-2 and Bcl-2 mRNA was identified by competitive RT-PCR. RESULTS: NS-398 inhibited cell proliferation and induced apoptosis in HepG2 in a concentration-dependent manner. DNA ploidy analysis showed that S phase cells were significantly decreased with NS-398 concentration increasing. The quiescent G₀/G₁ phase was accumulated with decreasing of Bcl-2 mRNA. Whereas NS-398 had no effect on the expression of COX-2 mRNA, no correlations were found between COX-2 mRNA and the HepG2 cell proliferation and apoptosis induced by NS-398 (r=0.056 and r=0.119, respectively). CONCLUSION: NS-398 significantly inhibits the proliferation and induces apoptosis in HepG2. Mechanisms may be involved in accumulation of quiescent G₀/G₁ phase and decrease in Bcl-2 mRNA expression, but independent to COX-2 mRNA expression.     

Evolutionary plasticity of monooxygenase-mediated resistance

The cytochrome P450 monooxygenases are an important metabolic system involved in the detoxification of xenobiotics, and are thus one of the major mechanisms by which insects evolve insecticide resistance. However, comparatively little is known about the evolutionary constraints of this insecticide resistance mechanism. We investigated the genetic basis of resistance in a strain of house fly (NG98) from Georgia, USA that had evolved 3700-fold resistance to the pyrethroid insecticide permethrin, and compared this to other permethrin resistant strains of house flies from the US and Japan. Resistance in NG98 was due to kdr on autosome 3 and monooxygenase-mediated resistance on autosomes 1, 2, and 5. These results indicate that the genes which evolve to produce monooxygenase-mediated resistance to permethrin are different between different populations, and that the P450 monooxygenases have some degree of plasticity in response to selection. Monooxygenase-mediated resistance appears to evolve using different P450s, and possibly different regulatory signals controlling P450 expression, even in strains selected with the same insecticide.     

旋毛虫T3223-6 cDNA的表达及鉴定

利用PCR技术将T3223-6 cDNA扩增克隆到原核表达载体pET-28a,将重组质粒转入克隆菌Nova-blue,提取质粒进行酶切和测序鉴定后转入表达菌BL21star(DE3).用1 mM IPTG诱导培养重组表达菌,对菌体裂解物进行SDS-PAGE分析,检测重组蛋白的表达情况.用旋毛虫感染猪血清和正常血清,通过western blotting检测重组蛋白的反应原性.结果表明:经IPTG诱导后重组转化菌的裂解产物出现44 kD左右的表达带,大小与理论值相符;western blotting检测结果显示重组蛋白可以被旋毛虫感染猪血清识别,具有反应原性.