医学微生物学双语教学研究

实施双语教学是适应我国高等教育国际化趋势的发展需要.结合本校实际,从采用英文原版教材进行医学微生物学双语教学必要性入手,对医学微生物学双语教学方式、教学评价、教学效果等进行了探讨,为我校双语教学模式的选择应用提供了一定的依据.     

用人工神神经网络法辨识发酵动力学模型参数

本文提出发酵动力学模型参数辩识的人工神经网络方法,并对几种典型的模型进行了具体尝试,结果表明,用这种方法估计参数效果极好。     

Processing of Cry1Ab δ-endotoxin from Bacillus thuringiensis by Manduca sexta and Spodoptera frugiperda midgut proteases: role in protoxin activation and toxin inactivation

Activation of Cry protoxins is carried out by midgut proteases. This process is important for toxicity and in some cases for specificity. Commercial proteases have been used for in vitro protoxin activation. In the case of Cry1A protoxins, trypsin digestion generates a toxic fragment of 60–65 kDa. Here, we have analyzed the in vitro and in vivo activation of Cry1Ab. We found differences in the processing of Cry1Ab protoxin by Manduca sexta and Spodoptera frugiperda midgut proteases as compared to trypsin. Midgut juice proteases produced two additional nicks at the N-terminal end removing helices α1 and α2a to produce a 58 kDa protein. A further cleavage within domain II splits the toxin into two fragments of 30 kDa. The resulting fragments were not separated, but instead coeluted with the 58 kDa monomer, in size-exclusion chromatography. To examine if this processing was involved in the activation or degradation of Cry1Ab toxin, binding, pore formation, and toxicity assays were performed. Pore formation assays showed that midgut juice treatment produced a more active toxin than trypsin treatment. In addition, it was determined that the α1 helix is dispensable for Cry1Ab activity. In contrast, the appearance of the 30 kDa fragments correlates with a decrease in pore formation and insecticidal activities. Our results suggest that the cleavage in domain II may be involved in toxin inactivation, and that the 30 kDa fragments are stable intermediates in the degradation pathway.     

鲤鲫人工多倍体谱系中同工酶和蛋白的基因表达

通过对红鲤、红鲫、镜鲤、鲤鲫杂种二倍体一代,二代,鲤鲫杂种三倍体,鲤鲫复合三倍体,鲤鲫杂种四倍体一代,二代的同工酶及蛋白电泳谱型和扫描数据分析表明,在鲤鲫人工多倍体谱系中,亲代的等位基因在杂交子代中共有四种表达模式;（1）两亲本基因在子代中共同表达,即共显表达;（2）父本的基因表达受到部分或完全的抑制,即母本的基因优先得到表达;（3）母本的基因表达受到抑制,父本的基因得到表达;（4）双亲本的基因表达均受到一定程度的抑制或都不表达。其中第一种表达模式是主要的模式。根据以上基因在杂交子代中的表达特点,可用同工酶和蛋白电泳图谱将鲤鲫人工多倍体谱系的各种生物型逐一加以区分。     

淡水育珠蚌外套膜表皮细胞分泌方式的研究

用光学显微镜、相差显微镜、透射电镜、扫描电镜对３种淡水育珠蚌外套膜表皮细胞的分泌方式进行了观察研究。几种显微镜观察对比研究的结果表明,外套膜细胞的分泌方式主要有小泡式的局部分泌;液流式、缺口式的顶浆分泌和巨浆分泌;分泌物排出时连同整个细胞一起排出和形成复层表皮后外层脱落的全浆分泌。内表皮细胞以小泡式和缺口式分泌为主;外表皮细胞多为液流式分泌和全浆分泌。内表皮分泌活动表现为持续性和连续性的特点,观察相邻细胞同时可见到各种不同的分泌时相;外表皮分泌活动呈节律性和区段性的特点,分泌活动旺盛的细胞区段和无分泌活动的区段相间出现。以上结果表明内、外表皮细胞在分泌方式及分泌物性质上均有显著差异,这种差异反映了内、外表皮功能上的不同。     

Catalytic properties and mode of action of endo-(1-->3)-beta-D-glucanase and beta-D-glucosidase from the marine mollusk Littorina kurila

A complex of the enzymes from the liver of the marine mollusk Littorina kurila that hydrolyzes laminaran was investigated. Two (1-->3)-beta-d-glucanases (G-I and G-II) were isolated. The molecular mass of G-I as estimated by gel-permeation chromatography and SDS-PAGE analysis was 32 and 40kDa, respectively. The G-II molecular mass according to SDS-PAGE analysis was about 200kDa. The pH optimum for both G-I and G-II was pH 5.4. The G-I had narrow substrate specificity and hydrolyzed only the (1-->3)-beta-d-glucosidic bonds in the mixed (1-->3),(1-->6)- and (1-->3),(1-->4)-beta-d-glucans down to glucose and glucooligosaccharides. This enzyme acted with retention of the anomeric configuration and catalyzed a transglycosylation reaction. G-I was classified as the glucan endo-(1-->3)-beta-d-glucosidase (EC 3.2.1.39). G-II exhibited both exo-glucanase and beta-d-glucoside activities. This enzyme released from the laminaran glucose as a single product, but retained the anomeric center configuration and possessed transglycosylation activity. The hydrolysis rate of glucooligosaccharides by G-I decreased with an increase of the substrate's degree of polymerization. In addition to (1-->3)-beta-d-glucanase activity, the enzyme had the ability to hydrolyze p-nitrophenyl beta-d-glucoside and beta-d-glucobioses: laminaribiose, gentiobiose, and cellobiose, with the rate ratio of 50:12:1. G-II may correspond to beta-d-glucoside glucohydrolase (EC 3.2.1.21).     

fMRI颞叶癫痫默认模式网络的研究

许多功能磁共振研究已经发现人脑的一些皮层区域在静息状态下出现共激活,这些区域形成连通的功能网络,称为"默认模式网络"。本文研究颞叶癫痫患者的默认模式网络,运用独立成分分析(Independent component analysis)分离出12例颞叶癫痫患者和12例正常对照的默认模式网络,进行组内分析得到两组被试的统计图,进行组间分析比较两组被试的默认模式网络的差异。结果表明默认模式网络均存在于颞叶癫痫患者和正常对照中,然而,在默认模式包含的网络中,颞叶癫痫患者前扣带回腹侧(ventral anterior cingulated cortex,vACC)、前额中分(medial prefrontal cortex,MPFC)、楔前叶(precuneus)、以及海马旁回区域比正常对照表现出代谢增强。这一结果有助于从脑功能的角度了解癫痫患者某些临床症状的发病机理,为今后癫痫诊治的发展提供一定的帮助。     

Design,synthesis and antifungal activity evaluation of isocryptolepine derivatives

In this paper, the nitrogen atom was inserted into the anthracycline system of the isocryptolepine nucleus to obtain the “Aza”-type structure benzo[4,5]imidazo[1,2-c] quinazoline. A series of “Aza”-type derivatives were designed, synthesized and evaluated for their antifungal activity against six plant fungi in vitro. Among all derivatives, compounds A-0, B-1 and B-2 showed significant antifungal activity against B. cinerea with the EC₅₀ values of 2.72 μg/mL, 5.90 μg/mL and 4.00 μg/mL, respectively. Compound A-2 had the highest activity against M. oryzae with the EC₅₀ values of 8.81 μg/mL, and compound A-1 demonstrated the most control efficacy against R. solani (EC₅₀, 6.27 μg/mL). Moreover, compound A-0 was selected to investigate the in vivo tests against B. cinerea and the results indicated that the preventative efficacy of it up to 72.80% at 100 μg/mL. Preliminary mechanism studies revealed that after treatment with A-0 at 5 µg/mL, the B. cinerea mycelia appeared curved, collapsed and the cell membrane integrity may be damaged. The reactive oxygen species production, mitochondrial membrane potential and nuclear morphometry of mycelia have been changed, and the membrane function and cell proliferation of mycelia were destroyed. Compounds A-0, A-1, B-1 and B-2 presented weaker toxicities against two cells lines than isocryptolepine. This study lays the foundation for the future development of isocryptolepine derivatives as environmentally friendly and safe agricultural fungicides.     

Spontaneous bacteriocin resistance in Listeria monocytogenes as a susceptibility screen for identifying different mechanisms of resistance and modes of action by bacteriocins of lactic acid bacteria

A practical system was devised for grouping bacteriocins of lactic acid bacteria (LAB) based on mode of action as determined by changes in inhibitory activity to spontaneously-acquired bacteriocin resistance (Bac^R). Wild type Listeria monocytogenes 39-2 was sensitive to five bacteriocins produced by 3 genera of LAB: pediocin PA-1 and pediocin Bac3 (Pediococcus), lacticin FS97 and lacticin FS56 (Lactococcus), and curvaticin FS47 (Lactobacillus). A spontaneous Bac^R derivative of L. monocytogenes 39-2 obtained by selective recovery against lacticin FS56 provided complete resistance to the bacteriocin made by Lactococcus lactis FS56. The lacticin FS56-resistant strain of L. monocyotgenes 39-2 was also cross-resistant to curvaticin FS47 and pediocin PA-1, but not to lacticin FS97 or pediocin Bac3. The same pattern of cross-resistance was also observed with Bac^R isolates obtained with L. monocytogenes Scott A-2. A spontaneous mutation that renders a strain cross-resistant to different bacteriocins indicates that they share a common mechanism of resistance due to similar modes of action of the bacteriocins. Spontaneous resistance was acquired to other bacteriocins (in aggregate) by following the same procedure against which the Bac^R strain was still sensitive. In subsequent challenge assays, mixtures of bacteriocins of different modes of action provided greater inhibition than mixtures of bacteriocins of the same mode of action (as determined by our screening method). This study identifies a methodical approach to classify bacteriocins into functional groups based on mechanism of resistance (i.e., mode of action) that could be used for identifying the best mixture of bacteriocins for use as biopreservatives.     

不同分娩方式对晚期早产儿肠道菌群的影响

目的 探讨不同分娩方式对晚期早产儿肠道菌群定植的影响。方法 以胎龄（周）为340/7～366/7的15例晚期早产儿为研究对象,根据分娩方式分为自然分娩组（n=8）和剖宫产组（n=7）。收集早产儿出生后3 d、7 d、14 d的粪便标本,应用高通量测序技术对细菌16S rRNA可变区中的V4区进行测序,分析肠道菌群多样性及组成结构。结果 （1）自然分娩组晚期早产儿粪便标本菌群多样性指数逐渐上升,剖宫产组的多样性指数较平稳,两组相比差异无统计学意义;（2）45份粪便标本中共检测出10个菌门,均以变形菌门、厚壁菌门、放线菌门和拟杆菌门为优势菌门,两组晚期早产儿生后变形菌门、拟杆菌门所占比例逐渐降低,厚壁菌门、放线菌门呈增多趋势。两组相比,剖宫产组7 d、14 d时拟杆菌门的相对丰度显著低于自然分娩组（Z=‒2.896,P=0.004;Z=‒2.120,P=0.040）,变形菌门相对丰度仅在7 d时显著高于自然分娩组（Z=‒2.190,P=0.030）;（3）两组研究对象中,除自然分娩组14 d时以双歧杆菌属为优势菌属外,余下均以肠杆菌属为优势菌属。相比于自然分娩组,在7 d时剖宫产组拟杆菌属所占比例显著降低（Z=‒2.806,P=0.005）,肠杆菌属所占比例显著升高（Z=‒2.199,P=0.030）。结论 剖宫产能显著影响婴儿早期肠道菌群的定植,降低肠道中早期拟杆菌的水平。