不同冷冻保藏条件对Glarea lozoyensis生长及产纽莫康定B_0能力的影响

棘白菌素类化合物纽莫康定B_0生产菌株Glarea lozoyensis,在其连续传代和发酵过程中都观察到菌株的变异,给科研、生产带来极大困扰。因此,采用低温冷冻保藏对G.lozoyensis Q1进行长期保存。采用单因素实验法研究了低温保护剂类型、保藏温度以及低温保护剂浓度对G.lozoyensis Q1菌体形态和发酵性能的影响。以作用于全细胞的80 g/L甘油和作用于细胞壁外的200 g/L PEG-6000作为低温保护剂,在-80℃保藏3个月后,纽莫康定B_0产量达到1.6 g/L,与保藏前基本一致。并且,当菌体保藏过后菌丝形态能维持一定褶皱的实验组产量均能保持在1 g/L以上,当表面趋于光滑时产量急剧下降,仅为初始的25%左右。结果表明,以甘油及PEG-6000作为冷冻保护剂,在-80℃条件下可实现G.lozoyensis Q1长期保藏。     

不同培养条件对猪卵母细胞IVM、IVF的影响

通过优化猪卵母细胞体外成熟、体外受精和胚胎体外发育体系,以进一步提高体外胚胎的生产效率和质量.研究了激素存在时间、不同激素和不同血清对猪卵母细胞体外成熟的影响;共培养体系、精卵作用时间、去除卵丘细胞的方法对猪体外受精及早期胚胎发育的影响.猪卵母细胞IVM培养48h,前24h内加入PMSG、hCG,后24h将其去除,卵母细胞总成熟率为79.54%;培养液添加15?S或15%NCS,卵母细胞成熟率分别为79.48%和74.81%;PMSG、HCG和E2配合使用后卵母细胞成熟率为81.42%.在IVF前用吹打法获得的卵裂率、桑椹胚率分别为37.89%和8.54%,精卵共孵育6h或8h的卵裂率(40.52%,37.24%)、桑椹胚率(8.42%,7.85%),以及用输卵管上皮细胞共培养所获得的卵裂率(40.84%)、桑椹胚率(9.53%)均显著高于其它各组.     

睫状神经营养因子对大鼠去神经骨骼肌的营养作用

目的：了解睫状神经营养因子（CNTF）对去神经引起的肌肉萎缩的治疗作用。方法：离断SD大鼠一侧坐骨神经,连续给予CNTF20d,观察肌肉湿重、蛋白含量、肌纤维横截面积、收缩性能和残肢程度。结果：①给予0.2mg/kg的CNTF,可使损务侧肌纤维横截面积增加35％,肌肉湿重增加38％,胫前肌总蛋白含量增加24％,腓长肌强直收缩强度提高40％,显著改善肢残程度;②0.2mg/kg的CNTF作用明显强于0.05mg/kg的CNTF;③此目鱼肌（慢肌）比伸趾长肌（快肌）对CNTF更敏感。结论：CNTF能显著改善成年大鼠坐骨神经离断后骨骼肌的萎缩和功能丧失,该效应的强弱与用药剂量和肌肉类型有关。     

早期人胚胎cDNA文库构建及目的基因筛选

收集受精后３、４和５周龄药物流产胚胎,用改良一步法提取总ＲＮＡ,ｏｌｉｇｏ（ｄＴ）纤维素柱纯化ｍＲＮＡ,逆转录合成一链ｃＤＮＡ,完成二链ｃＤＮＡ的合成后,经碱变性电泳检测,合成ｃＤＮＡ的大小为０．４～９．０ｋｂ之间,且主要集中在１．０～２．０ｋｂ。除去多余的接头,收集大于４００ｂｐ的ｃＤＮＡ片段,与载体ｐＳＰＯＲＴ１和和γＺｉｐＬｏｘ连接,分别得到３、４、５周龄人胚胎质粒文加和噬菌体文库。另外,采     

药用植物栀子的组织培养

栀子(Gardenia jasminoides)为药用木本植物。以栀子果皮、种子团和种子为外植体,研究不同激素配比及不同培养方式对愈伤组织诱导和芽分化的影响。研究结果表明,培养基成分为MS+0.5 mg·L–12,4-D+0.25 mg·L–16-BA较适宜果皮和种子愈伤组织的诱导,诱导率分别为83.3%和88.5%;培养基成分为MS+1.0 mg·L–12,4-D+1.0 mg·L–16-BA较适宜种子团愈伤组织的诱导,诱导率为78.1%。3种外植体诱导的愈伤组织中,只有种子愈伤组织能通过液体培养分化出芽;TDZ对芽分化有明显的促进作用;最佳的芽分化培养基为MS+0.05 mg·L–1NAA+0.10 mg·L–1TDZ,其愈伤组织分化率为8.75%。该研究以栀子种子为外植体,并获得了再生植株,为药用植物栀子转基因体系的建立奠定了基础。     

Microbial 2,3-butanediol production: a state-of-the-art review

2,3-Butanediol is a promising bulk chemical due to its extensive industry applications. The state-of-the-art nature of microbial 2,3-butanediol production is reviewed in this paper. Various strategies for efficient and economical microbial 2,3-butanediol production, including strain improvement, substrate alternation, and process development, are reviewed and compared with regard to their pros and cons. This review also summarizes value added derivatives of biologically produced 2,3-butanediol and different strategies for downstream processing. The future prospects of microbial 2,3-butanediol production are discussed in light of the current progress, challenges, and trends in this field. Guidelines for future studies are also proposed.     

A combined A431 cell membrane chromatography and online high performance liquid chromatography/mass spectrometry method for screening compounds from total alkaloid of Radix Caulophylli acting on the human EGFR

We have developed an online analytical method that combines A431 cell membrane chromatography (A431/CMC) with high performance liquid chromatography and mass spectrometry (LC/MS) for identifying active components from Radix Caulophylli acting on human EGFR. Retention fractions on A431/CMC model were captured onto an enrichment column and the components were directly analyzed by combining a 10-port column switcher with an LC/MS system for separation and preliminary identification. Using Sorafenib tosylate as a positive control, taspine and caulophine from Radix Caulophylli were identified as the active molecules which could act on the EGFR. This A431/CMC-online-LC/MS method can be applied for screening active components acting on EGFR from traditional Chinese medicines exemplified by Radix Caulophylli and will be of great utility in drug discovery using natural medicinal herbs as a source of novel compounds.