卷曲乳杆菌对三种结肠上皮细胞的粘附性

目的 研究卷曲乳杆菌对3种结肠上皮细胞（SW480细胞、SW620细胞、LOVO细胞）的粘附性。方法 将处于对数生长期的卷曲乳杆菌A7分别与SW480细胞、SW620细胞、LOVO细胞进行体外粘附试验,革兰染色后显微镜观察卷曲乳杆菌A7对3种结肠上皮细胞的粘附结果并计数。结果 卷曲乳杆菌A7对3种结肠上皮细胞的粘附均具有显著性,其中对于SW480细胞和LOVO细胞的粘附性明显高于SW620细胞。结论 卷曲乳杆菌A7对SW480细胞、SW620细胞、LOVO细胞均具有较强的粘附性,提示该菌株有望成为肠道益生菌的新成员。     

甲硝唑与乳杆菌活菌制剂对细菌性阴道病患者阴道微生物群影响的对比性研究

目的对300例细菌性阴道病（Bacterial Vaginosis,BV）患者随机使用甲硝唑（100例）、乳杆菌活菌制剂定君生（100例）及甲硝唑联合定君生（100例）,观察乳杆菌在阴道内定植情况及用药前后阴道内微生物多样性的变化,为临床治疗细菌性阴道病提供实验依据。方法分别提取300例BV患者用药前和用药后的阴道微生物总基因组DNA,以德氏乳杆菌16SrDNA特异性引物进行PCR,检测患者用药后德氏乳杆菌在阴道内的定植情况。以细菌16SrRNAV3区通用引物进行PCR,扩增产物进行变性梯度凝胶电泳（DGGE）,对所得指纹图谱进行分析,研究用药对微生态变化的影响状况。结果甲硝唑治疗组用药前后均没有德氏乳杆菌检出;定君生治疗组用药后德氏乳杆菌检出率为54％;甲硝唑联合定君生治疗组用药后德氏乳杆菌检出率为76％。甲硝唑治疗组用药后微生物多样性显著降低,微生物群结构失衡;定君生治疗组用药后微生物多样性有所降低,微生物群结构无显著差异;甲硝唑联合定君生治疗组用药后微生物多样性降低,但较前两组有所增加,微生物群结构差异无统计学意义。结论甲硝唑显著降低BV患者阴道微生物多样性,引起微生态失衡;定君生对BV患者阴道微生态的影响明显小于甲硝唑;定君生的使用有利于甲硝唑f预后阴道微生物多样性的恢复和微生态平衡的重建。     

我国登革3型病毒广西80-2株基因组全序列分析

对我国登革 3型病毒 80 2株基因组进行全序列测定 ,为了解其基因组结构与功能的关系提供依据 .根据登革 3型病毒H87株的序列设计并合成引物 ,应用RT PCR和RACE法 ,对 80 2株基因组RNA进行扩增、克隆测序后获得我国登革 3型病毒广西株基因组序列 .该株病毒基因组全长10 696nt ,不含poly(A)尾 ,4种碱基数分别为A :3 4 3 7,C :2 2 15,G :2 773 ,U :2 2 71.包含一个读码框架 ,自 95至 10 2 67位 ,共 10 170个碱基 ,编码 3 3 90个氨基酸 ,5′和 3′非编码区长度分别为 94nt和4 3 2nt.与H 87株比较 ,核苷酸和氨基酸序列同源性均在 99%以上 ,有 2 8个碱基发生改变 ,其中 2 6个碱基突变发生在读码框架内 ,碱基转换 18个 ,颠换 10个 ;碱基突变引起 14个氨基酸的改变 .80 2株与H87株病毒的基因组全序列同源性高 ,变异度小 .     

狂犬病病毒Evelyn- Rokitnicki-Abelseth疫苗株反向遗传系统的建立

摘要：【目的】建立狂犬病毒的反向遗传系统,为研制不含狂犬病病毒致病性的新型安全高效的狂犬疫苗提供技术依据。【方法】本研究采用反向遗传学方法和分子克隆技术,建立了狂犬病病毒Evelyn- Rokitnicki-Abelseth (ERA)疫苗株的CMV/ T7、T7启动子病毒拯救系统,构建了表达N、P、L蛋白的辅助质粒。【结果】成功拯救出野生型病毒rERA-VC,在Vero细胞上的生长动力学特性与父母本ERA 相同,第三代可在Vero细胞上可获得很高的生长滴度。【结论】建立了狂犬病病毒的反向遗传系统,拯救出的野生型病毒生物学特性与父母本相同。     

近交系小鼠微卫星DNA多态性的研究

随机选择位于小鼠不同染色体上的微卫星引物42对,用PCR技术对C3H、C57、BALB/c、DBA、TA2、T739、B615、BACB/c－nu－nu和SCID等9种实验室常用近交系小鼠微卫星DNA多态性进行了研究。结果显示有信息的40对引物中,9种近交系小鼠在各基因座上均出现一条清晰条带,28个基因座表现为多态性。其中D3Mit22、D7Nds1、D11Mit12、D12Nds2、D15Mit17、D16Mit3、D16Mit4基因座表现为显著多态性。T739、B615和TA2的遗传背景相近,其相似系数分别为90％和85％;其次为TA2、SCID和B615,其相似系数分别为80％和82.5％。结果表明所检测的小鼠符合近交要求,筛选出的引物能典型地反映9个近交系小鼠的品系特异性和遗传背景,可用于常规检测小鼠品系来源和遗传背景等。 Abstract：Forty-two microsatellites DNA loci on different chromosomes in nine kinds of inbred strain mice including C3H,C57,BALB/c,DBA,TA2,T739,B615,BALB/c-nu-nu and SCID were investigated by PCR analysis.It showed that all these mice tested display single allelic gene band with forty pairs of informative primers.Twenty-eight loci are polymorphisms,among which the polymorphisms of D3Mit22,D7Nds1,D11Mit12,D12Nds2,D15Mit17,D16Mit3,and D16Mit4 loci are significant.The genetic background of T739 was similarity with that of B615 and TA2 ,the similarity indices were 90％ and 85％ respectively;and that of TA2 was similarity with SCID and B615,the similarity indices were 80％ and 82.5％.These results suggest that these mice tested meet the request of inbred strain.Screened primers showing marked polymorphisms topically reflect the speciality of strains and genetic backgrounds,which could be used in determining the strains origin and genetic background of mice.     

苹果属皱叶矮生型株系及其亲本和后代实生苗的过氧化物酶同工酶特性研究

以母本平邑甜茶1株、父本扎矮山定子1株、皱叶矮生型株系(F1)6株及其自然授粉后代实生苗(F2)15株为试材,采用聚丙烯酰胺凝胶电泳法研究叶片过氧化物酶同工酶谱的异同。结果表明:皱叶矮生型株系叶片的过氧化物酶同工酶酶谱共有10条酶带,R f值范围0.18~0.90,各植株间的酶带数量基本一致,只有强弱不同,与平邑甜茶酶谱的差异也很小。在皱叶矮生型株系的后代实生苗叶片中,除15 a单株有16条酶带外,其他实生苗的过氧化物酶同工酶酶谱约有10条酶带,但株间差异明显,R f值范围0.18~0.84。不同树龄平邑甜茶叶片的酶谱一致性较好。     

The Bacterial Peptide Pheromone Plantaricin A Permeabilizes Cancerous, but not Normal, Rat Pituitary Cells and Differentiates between the Outer and Inner Membrane Leaflet

Plantaricin A (PlnA) is a 26-mer peptide pheromone with membrane-permeabilizing, strain-specific antibacterial activity, produced by Lactobacillus plantarum C11. We investigated the membrane-permeabilizing effects of PlnA on cultured cancerous and normal rat anterior pituitary cells using patch-clamp techniques and microfluorometry (fura-2). Cancerous cells displayed massive permeabilization within 5 s after exposure to 10–100 μm PlnA. The membrane depolarized to nearly 0 mV, and the membrane resistance decreased to a mere fraction of the initial value after less than 1 min. In outside-out membrane patches, 10 μm PlnA induced membrane currents reversing at 0 mV, which is compatible with an unspecific conductance increase. The d and l forms of the peptide had similar potency, indicating a nonchiral mechanism for the membrane-permeabilizing effect. Surprisingly, inside-out patches were insensitive to 1 mm PlnA. Primary cultures of normal rat anterior pituitary cells were also insensitive to the peptide. Thus, PlnA differentiates between plasma membranes and membrane leaflets. Microfluorometric recordings of [Ca²⁺] _i and cytosolic concentration of fluorochrome verified the rapid permeabilizing effect of PlnA on cancerous cells and the insensitivity of normal pituitary cells.     

环丙沙星预先处理牙鲆消化道对干酪乳杆菌定植的影响

利用1株干酪乳杆菌,通过实验研究用环丙沙星预先处理牙鲆消化道后乳杆菌的定植和演替规律。在投喂含有1.2×10^9CFU/g乳杆菌的饲料5 d后,消化道定植的乳杆菌超过106CFU/g,其后维持在10^6～10^8CFU/g动态平衡中。同时随着乳杆菌的投喂,不经环丙沙星预先处理牙鲆消化道的正常组,鱼消化道的弧菌数从10^7～8CFU/g降低到10^6CFU/g左右;而经环丙沙星预先处理牙鲆消化道的药饵组,鱼胃、小肠和盲囊的弧菌数则是先增加,然后显著下降。停喂乳杆菌7 d后,2个实验组鱼消化道的乳杆菌均从10^5～6CFU/g下降到10^4CFU/g,干酪乳杆菌正常组鱼盲囊中弧菌从10^5CFU/g回升到10^6CFU/g,胃和小肠中弧菌数量基本不变。干酪乳杆菌药饵组则有所不同：除胃中弧菌数量则基本不变外,盲囊和小肠中弧菌数量继续下降,其原因有待进一步研究。实验结果表明,干酪乳杆菌能在牙鲆消化道内定植,而用预先处理牙鲆消化道后,更有利于乳杆菌的定植;乳杆菌的投喂和定植,使牙鲆消化道中的弧菌数量明显下降。     

唾液酸醛缩酶基因工程菌的构建和表达*

唾液酸(NANA)是一族神经氨酸类衍生物,它处于许多糖蛋白的寡糖链的非还原末端,具有重要的生理功能和药用价值。由于唾液酸的常规化学合成分离方法复杂,难以大量制备,因此价格很贵。而用酶法合成唾液酸,即在唾液酸醛缩酶(ALD)作用下,以丙酮酸钠和NI乙酰甘露糖胺为底物合成唾液酸．则原料便宜,步骤简单、产率高,且适合工业化生产。但唾液酸醛缩酶是一个诱导酶,只能在以唾液酸为唯一碳源的培养基下才能生成,这就大大影响了它的应用。因此,我们构建了产该酶的工程菌,为唾液酸作为药物和原料药的大量应用打下了基础。     

Burkholderia phenoliruptrix sp. nov., to accommodate the 2,4,5-trichlorophenoxyacetic acid and halophenol-degrading strain AC1100

Strain AC1100 is well-known for its ability to degrade a variety of recalcitrant xenobiotics, including 2,4,5-trichlorophenoxyacetic acid. We performed a polyphasic-taxonomic study to determine its taxonomic position. The G+C content of strain AC1100 was 62.6 mol%. On the basis of 16S rRNA gene sequence similarity, strain AC1100 belonged to the b-Proteobacteria and was most closely related to Burkholderia fungorum (98.3% similarity). DNA-DNA hybridisations, comparison of protein profiles, cellular fatty acid analysis and biochemical tests allowed genotypic and phenotypic differentiation of strain AC1100 from other Burkholderia species. Our data show that strain AC1100 represents a novel species for which the name Burkholderia phenoliruptrix sp. nov. is proposed. The type strain is AC1100^T (= LMG 22037^T = CCUG 48558^T).