Ribosomal DNA internal transcribed spacer sequences do not support the species status of Ampelomyces quisqualis, a hyperparasite of powdery mildew fungi

Phylogenetic relationships among Ampelomyces isolates, pycnidial hyperparasites and biological control agents of powdery mildews, were inferred from internal transcribed spacer (ITS) sequences of the ribosomal DNA (rDNA). Currently, these hyperparasites are considered to be a single species, A. quisqualis, despite observed morphological and cultural differences. Ten Ampelomyces isolates, representing seven previously defined ITS RFLP groups, were sequenced and analyzed. Sequence-divergence values among isolates belonging to different RFLP groups ranged from 4.3 to 22.4%, suggesting that these isolates may represent different taxa. When Ampelomyces ITS sequences were analyzed by cladistic methods with the sequences of other ascomycetous fungi, they formed two lineages in the Dothideales. Slow-growing Ampelomyces isolates formed a clade with Leptosphaeria microscopica and L. nodorum, whereas fast-growing Ampelomyces isolates formed a clade with Epicoccum nigrum. Sequence-divergence values between these two clades ranged from 17.3 to 22.4%, suggesting that the taxa in the two clades are not closely related and possibly not congeneric. The data presented here indicate that the identification of `A. quisqualis' isolates used in biological control experiments should be re-evaluated. Received: 10 March 1997 / Accepted: 13 February 1998     

Genetic analysis of DNA fingerprints and virulences in Erysiphe graminis f.sp. hordei

A DNA probe, E9, which has been used extensively in population genetic studies of the barley powdery mildew pathogen, Erysiphe graminis f.sp. hordei, was shown to be homologous to dispersed sequences in the genome of this fungus. In a cross of the isolates CC52 and DH14, fragments with homology to E9 mapped to six clusters of loci. Avirulences matching five resistance genes in barley were controlled by single genes, in accordance with the gene-for-gene hypothesis, while avirulence matching a sixth resistance gene, Mla13, was controlled by two genes. A gene which controls the response to a fungicide, ethirimol, was not linked to any other gene. In all, seven linkage groups, comprising 22 loci, were detected. The results indicate that E9 can be used to identify members of a clone of E.g. f.sp. hordei, but should not be used for quantitative population genetic analysis.     

HPLC法测定银杏叶粉针中总黄酮醇苷和萜类内酯

目的采用高效液相色谱法测定银杏叶粉针中总黄酮醇苷和总萜类内酯。方法黄酮醇苷水解后,采用HPLC-UV法测定;萜类内酯用醋酸乙酯萃取后采用HPLC-ELSD法测定。结果槲皮素、山柰素、异鼠李素分别在5.104～51.04、5.000～50.00、1.38～13.8μg/mL呈良好线性关系,平均回收率分别为99.34%、99.25%、101.2%(n=5);银杏内酯A、B、C及白果内酯分别在0.415～2.075、0.402～2.010、0.426～2.130、0.803～4.015mg/mL呈良好线性关系,平均回收率分别为99.28%、100.1%、101.0%、99.31%(n=5)。结论本实验采用的方法简便快速,结果准确,重现性好,可用于银杏叶粉针中黄酮和内酯类成分的质量控制。     

促肝细胞生长素注射液的临床前实验研究

目的 对促肝细胞生长素注射液(Injection of PHGF)进行临床前实验研究,为新剂型的研究提供依据。方法 3批促肝细胞生长素注射液在室温下进行加速实验,用高效液相色谱仪对其组分分析、用H3-掺入法和E花环法对3批注射液的生物学和免疫学活性研究;新西兰兔耳缘静脉注射促肝细胞生长素注射液,肉眼观察和显微镜检注射部位血管及其周围组织的病理改变,研究促肝细胞生长素注射液的血管刺激性;以生理盐水为阴性对照,注射用水为阳性对照,加入适量2％红细胞悬液,促肝细胞生长素注射液为实验组,进行溶血实验研究;常规方法进行过敏实验研究。结果 室温下加速实验6个月的3批促肝细胞生长素注射液与粉针剂的组分一致;生物学活性分别为8.6、5.6、5.0,免疫学活性分别为15.7％、12.9％、12.4％,与3批粉针剂比较均P>0.05,结果无显著性差异;促肝细胞生长素注射液无血管刺激性;无溶血现象;过敏实验结果符合规定。结论 促肝细胞生长素注射液室温下放置6个月稳定性好,无血管刺激性,不溶血,不致敏。     

Pathogen-derived nitric oxide influences formation of the appressorium infection structure in the phytopathogenic fungus Blumeria graminis

Nitric oxide (NO) is an important signal in plant resistance to pathogens. Here we report that NO is also generated by Blumeria graminis f.sp. hordei as a pathogenesis determinant on barley. Infection by B. graminis f.sp. hordei is dependent on appressorium formation in order to penetrate the host. Using fluorescent dye diaminofluorescein-2 diacetate (DAF-2DA) and confocal laser scanning microscopy, transient NO generation was detected within the B. graminis f.sp. hordei appressorium during its maturation. To confirm that NO was indeed being measured, DAF-2DA fluorescence was suppressed using a NO scavenger and a mammalian NO synthase inhibitor. Both chemicals affected the number of appressorial lobes produced by the fungus. These data indicate that NO plays a key role in formation of B. graminis f.sp. hordei appressoria.     

注射用葛根素粉针与常用药物在4种输液中的稳定性考察

目的模拟临床用量,考察注射用葛根素粉针与常用药物在不同输液中的稳定性。方法通过观察外观、pH、含量等方法,判断葛根素分别与氯化钾、维生素B6、西米替丁、三磷酸腺苷二钠（ATP）、辅酶A配伍后0,1,2,4,6h时在不同输液中的稳定性。结果与结论葛根素粉针分别与维生素B6、氯化钾、西米替丁、ATP在5％葡萄糖注射液中配伍,pH会发生显著性变化,应谨慎配伍;其他配伍液外观、pH、含量无显著性改变,可配伍应用。     

中药材养护片剂防虫防霉试验初报

两种中药材养护片剂对5种中药材饮片的防虫霉效果:对千年健和苦杏仁均有明显防虫作用,虫蛀率分别比对照减少94.95％和95.55％,对款冬花也有一定的防霉作用。     

赛百粉针对难治性肾病综合征凝血/纤溶系统的影响

目的:探讨赛百粉针对难治性肾病综合征(RNS)凝血/纤溶系统的影响.方法:30例RNS随机分为治疗组与对照组,在常规治疗基础上,治疗组加用纤溶酶制剂赛百粉针,对照组加用低分子肝素钙速碧林注射液,比较两组治疗前后24 h尿蛋白、组织型纤溶酶原激活物(t-PA)、纤溶酶原激活物抑制物1(PAI-1)活性及血浆纤维蛋白原(Fib),D-二聚体(D-D)、纤溶酶原(PLG)含量、部分凝血活酶时间(APTT)、凝血酶原时间(PT)的变化.结果:治疗1个疗程后,治疗组与对照组尿蛋白均显著下降(P≤0.001),两组间治疗前、后比较均无差异(P＞0.05);t-PA、PLG治疗前治疗组与对照组均较健康组明显下降(P＜0.005),PAI-1、D-D、Fib明显升高(P≤0.000),PT、APTT显著缩短(P≤0.000),两组间比较均无差异(P＞0.05).t-PA、PLG治疗后治疗组与对照组均较治疗前显著升高(P＜0.05),PAI-1、Fib、D-D显著下降(P≤0.005),除t-PA对照组升高更显著(P=0.018),PAI-1两组比较无统计学差异外(P＞0.05),D-D、Fib、PLG治疗组变化较对照组更显著(P≤0.005).治疗后两组PT延长均无统计学意义(P＞0.05),APTT对照组显著延长(P=0.021),治疗组无显著延长(P=0.155).结论:赛百粉针能显著改善RNS凝血/纤溶异常,减少蛋白尿,有助于RNS的缓解.     

HPLC法测定银杏叶粉针中总黄酮醇苷和萜类内酯

目的采用高效液相色谱法测定银杏叶粉针中总黄酮醇苷和总萜类内酯。方法黄酮醇苷水解后,采用HPLC-UV法测定;萜类内酯用醋酸乙酯萃取后采用HPLC-ELSD法测定。结果槲皮素、山柰素、异鼠李素分别在5.104～51.04、5.000～50.00、1.38～13.8μg/mL呈良好线性关系,平均回收率分别为99.34%、99.25%、101.2%(n=5);银杏内酯A、B、C及白果内酯分别在0.415～2.075、0.402～2.010、0.426～2.130、0.803～4.015mg/mL呈良好线性关系,平均回收率分别为99.28%、100.1%、101.0%、99.31%(n=5)。结论本实验采用的方法简便快速,结果准确,重现性好,可用于银杏叶粉针中黄酮和内酯类成分的质量控制。     

高效液相色谱-电化学法测定双黄连粉针中黄芩甙和黄芩素的含量

采用高效液相色谱法,以电化学检测测定双黄连粉针中黄芩甙和黄芩素的含量。流动相0.1mol/L磷酸二氢钾-乙酸四氢呋喃(1000∶100∶190),色谱柱ERC-ODS-1161(6mm×100mm),检测电压+650mV,柱温55℃,流速0.8ml/min。结果:本法的最小检测限为25ng和50ng,平均回收率为100.1%和99.7%,重现性好。本法测定快速、简便、准确,适用于含黄芩甙和黄芩素的中药材和制剂的含量测定。