青霉素V钾滴丸的研制

为减少青霉素V钾的水解并提高其疗效，用聚乙二醇400和聚乙二醇6000为基质制备青霉素V钾滴丸。体外抑菌试验表明，青霉素v钾滴丸抑菌作用略强于人药片剂。     

丝核菌细胞核染色技术的研究

比较了5种不同的染色方法对丝核菌（Rhizoctonia)菌株细胞核的染色效果。结果表明，番红O－KOH的染色效果最好，Giemsa、苯胺蓝－甘油次之，染色效果最差的是苯胺蓝－乳酚油及曲利苯蓝－乳酚油。用番红O－KOH染色方法对分离自13种作物的57个丝核菌菌株进行细胞核染色，其中3个菌株为双核丝核菌，其余54个菌株为多核丝核菌。     

表达大肠杆菌K88ac-ST1-LTB融合蛋白工程菌株的免疫原性研究

已构建的能表达大肠杆菌K88ac-ST1-LTB融合蛋白的工程菌株BL21（DE3）（pXKST3LT5)及其表达产物经动物试验证实没有毒性反应。用从IPTG诱导的工程菌中提取的包涵体或经甲醛灭活的工程菌制成抗原，免疫小鼠，结果免疫小鼠至少能抵抗2MLD的大肠杆菌强毒株C83902（K88ac,ST^ ,L^ )的攻击，用提取的包涵体免疫家兔后，采集的血清能够中和天然ST1的毒性，这表明构建的工程菌株BL21（DE3）（pXKST3LT5)可以作为预防幼畜大肠杆菌性腹泻基因工程菌苗的候选株。     

The effect of some Frescon analogs on the aquatic snail,Lymnaea stagnalis: II. Correlation of neurotoxicity to Molluscicidal effectiveness

Although the molluscicide Frescon is a strong neurotoxin to the Lymnaea stagnalis central nervous system in vitro, it is probable that the exposure of the whole animal to this molluscicide fails to result in central nervous system abnormalities: Frescon does not appear to reach the brain in sufficient quantity to disrupt its normal activity. However, only those Frescon analogs found to be neurotoxic were molluscicidal, suggesting some related mode, if not site, of action. Frescon and its analogs may act by affecting excitable tissues other than the nervous system (e.g., the snail musculature) by altering certain functional and/or structural membrane properties.     

Antisense drug targeting with VEGF mRNA designed by computer aid and inhibitory effects on K562 cell line in vitro

AIM:The effective antisense sequences targeted VEGF mRNA with computer software would be screened and designed, and effect of them on growth K562 cells and protein expression of VEGF were studied with experiments.METHODS:Seven antisense sequences were selected and synthesized, which consisted of 18-20 deoxynucleotide acid and were modified with phosphorothioate, according to principle of low free energy of overall △G₃₇ Overall. Cell growth was assayed by trypan blue dye exclusion assay and level of VEGF protien in the media was determined by ELISA.RESULTS:Six of seven sequences were capable of inhibing growth of K562 cells and downregulating the VEGF protein expression significantly, compared with Scrambed control group. It was found that there was a close correlation between low level of overall △G₃₇ and antisense effectiveness (r=0.887,P<0.01).CONCLUSION:VEGF mRNA antisense oliogdeoxynucleotides, which were designed by computer software of RNAstructure, were able to inhibit growth of K562 and its protein expression. The VEGF mRNA may be new target attached by drugs. At same time, the computer aided design is useful methods to obtain the effective antisense.     

Protective effect of onychin on the human umbilical vein endothelial cells injured by menadione

AIM: To investigate the protective effect of onychin on the endothelial cells injured by oxidative stress. METHODS: The injured model was established by endothelial cells treated with menadione. The growth inhibitory rate of endothelial cell was determined by MTT assay; NO₂^-／NO₃^- concentration in the medium was determined by nitrate reductase assay; eNOS and caveolin-1 protein levels were determined by Western blot. RESULTS: Onychin significantly decreased the growth inhibitory rate of endothelial cells injured by menadione, increased NO₂^-／NO₃^- concentration in the medium and eNOS activity and up-regulated caveolin-1 expression. CONCLUSION: Onychin possesses a protective effect against endothelial cell injury induced by menadione via caveolin-1/eNOS pathway.     

Bcl-2 antisense oligodeoxynucleotide increases the sensitivity of HL60 and K562 cells to daunorubicin

AIM:To investigate whether the bcl-2 antisense oligonucleotide increases the sensitivity of HL60 and K562 cell lines to daunorubicin.METHODS:IC50 for HL60 and K562 was determined with MTT method, the expression levels of Bcl-2 protein were assayed by immunofluorescence using fluoresce isothiocyanate labeling. In addition, apoptosis was detected by morphological observation and flow cytometric analysis of DNA fragmentation.RESULTS:It was found that the two oligonucleotides directed against the coding region and the translation initiation of bcl-2 mRNA, combined respectively with daunorubicin, inhibited expression of bcl-2 protein, increased apoptosis in HL60 and K562 cells, and decreased IC50 of daunorubicin significantly (P＜0.05). Compared to the antisense oligonucleotide directed against the translation initiation of bcl-2 mRNA, the antisense oligonucleotide directed against the coding region showed stronger effects in the aspects of increasing the sensitivity of HL60 cells to daunorubicin (P＜0.05).CONCLUSIONS:These two antisense sequences in the translation initiation and the coding region of bcl-2 mRNA increased the sensitivity of HL60 and K562 cell lines to daunorubicin in a sequence-specific manner.     

Effect of Liucha extract on tumor K562 cell growth

AIM:To investigate the effects of Liucha extract on the growth of tumor cells in vitro and its possible mechanism.METHODS:The capability of colony forming of human leukemia K562 cell in vitro and the cells metabolism were studied by semi-solid agar culture and MTT staining. Then , the changes in morphology in the tumor cells were examined using electronic microscope.RESULTS:Semi-solid agar culture and MTT colorimetric analysis showed that Liucha extrats could significantly inhibit the growth of the tumor cells and their capability of colony forming. Also,under the electronic microscope,it was found that the tumor K562 cell had a narrower perinuclear space,condensation of chromatin and an enlarged mitochondria , in which the cristase disappeared.CONCLUSION:The extract from Liucha possesses an inhibitory effect on K562 cell growth in vitro through affecting the metabolism of the tumor cells.     

Development of Stem-base Pathogens on Different Cultivars of Winter Wheat Determined by Quantitative PCR

The progress of development of stem-base pathogens in crops of second winter wheat was plotted in nine experiments in three years. The amount of each pathogen present was determined by quantitative PCR. Where Tapesia yallundae was present in quantifiable amounts, it usually developed earlier than the other eyespot pathogen, T. acuformis. Both species were usually present in greater amounts on cultivars which are more susceptible to eyespot. The sharp eyespot pathogen, Rhizoctonia cerealis, developed more erratically than either of the Tapesia spp. and there were no consistent effects on different cultivars. Fusarium spp., the cause of brown foot rot, were rarely present in quantifiable amounts, but Microdochium nivale was usually present as one or both of the varieties nivale and majus. Late-season (after anthesis) decreases in M. nivale suggest that any brown foot rot symptoms attributable to this fungus would have fully developed earlier. Cultivar differences in amounts of M. nivale were most clear in stems during internode extension and when relatively large amounts of DNA were present. Such differences approximately reflected eyespot susceptibility, cv. Soissons containing most and cv. Lynx containing least DNA. The results emphasise the difficulty in relating diagnoses, by quantitative PCR or other means, at early growth stages when decisions to apply fungicides against stem-base disease are made, to later disease severity.