益生菌Lactobacillus casei Zhang固态发酵条件的优化

利用一株分离自传统发酵酸马奶中的益生干酪乳杆菌（Lactobacillus casei Zhang）进行固态发酵（Solid State Fermentation,SSF）。以发酵物中的活菌数为主要指标,采用九因素四水平（L32（4^9））的正交试验优化固态发酵培养基,并在优化的培养基基础上研究不同的初始含水量及培养时间对Lactobacillus casei Zhang活菌数的影响。实验结果表明,在固态发酵培养基组成为4g豆粕、5g麸皮、0．6g乳清粉、0．3g葡萄糖、0．3g碳酸钙、0．02g硫酸铵、0．01g硫酸镁,初始含水量为55％的优化条件下,37℃发酵60h,发酵物中Lactobacillus casei Zhang活菌数可达到4．08×10^10CFU／g。     

FCGR2B and FCRLB Gene Polymorphisms Associated with IgA Nephropathy

Background

IgA nephropathy (IgAN) is a complex syndrome characterized by deposition of IgA and IgA containing immune complexes (ICs) composed of IgG and complement C3 proteins in the mesangial area of glomeruli. The low-affinity receptors for the Fc region of IgG (FcγRs) are involved in autoantibody/immune complex-induced organ injury as well as ICs clearance. The aim of the study was to associate multiple polymorphisms within FCGR gene locus with IgAN in a large Chinese cohort.

Patients and Methods

60 single nucleotide polymorphisms (SNPs) spanning a 400 kb range within FCGR gene locus were analyzed in 2100 DNA samples from patients with biopsy proven IgAN and healthy age- and sex-matched controls from the same population in Chinese.

Results

Among the 60 SNPs investigated, 15 gene polymorphisms within FCGR gene locus (25%) were associated with susceptibility to IgAN. The most significantly associated SNPs within individual genes were FCGR2B rs12118043 (p = 8.74*10⁻³, OR 0.76, 95% CI 0.62–0.93), and FCRLB rs4657093 (p = 2.28*10⁻³, OR 0.77, 95% CI 0.65–0.91). Both conditional analysis and linkage disequilibrium analysis suggested they were independent signals associated with IgAN. Associations between FCGR2B rs12118043 and proteinuria (p = 3.65×10⁻²) as well as gross hematuria (p = 4.53×10⁻²), between FCRLB rs4657093 and levels of serum creatinine (p = 2.67×10⁻²) as well as eGFR (p = 5.41*10⁻³) were also observed. Electronic cis-expression quantative trait loci analysis supported their possible functional significance, with protective genotypes correlating lower gene expressions.

Conclusion

Our data from genetic associations and expression associations revealed potentially pathogenic roles of Fc receptor gene polymorphisms in IgAN.     

平贝母传粉生物学及繁育系统的研究

用TTC法测定花粉的活力,用联苯胺-过氧化氢法测定柱头的可授性,通过实地观测访花者、套袋实验、花粉-胚珠比(P/O比)的估算及花粉粒形态的观察研究了平贝母的传粉特性及繁育系统类型.结果表明,平贝母的花粉活力在开花后2～5 d可达90%以上,6 d后有所减弱,10 d后仍有71.3%的花粉保持活力.柱头的可授期约持续8 d,在开花后3～4 d柱头的可授性最强,5 d以后可授性逐渐减弱.平贝母的繁育系统是以自花传粉为主,虫媒异花传粉为辅的兼性自交.研究结果为平贝母的品种选育、杂交育种、引种栽培及物种保育提供了理论和技术依据,为深入研究贝母属植物的演化提供了生殖生态学依据.     

Ad-Ang1-IRES-VEGF165腺病毒载体的构建及其促血管形成作用

目的:构建人血管生成素1(Ang1)和血管内皮生长因子VEGF165 (VEGF165)的共表达腺病毒载体Ad-Ang1-IRES-VEGF165(简称Ad-AV),为研究Ad-AV转基因细胞表达产物血管诱生活性提供实验依据。方法:采用IRES介导的Ang1和VEGF165双基因腺病毒共表达模式,通过常规的基因克隆和重组技术,构建Ad-AV双基因共表达腺病毒载体,经感染人胚肾QBI-293A细胞(293A)进行扩增和效价测定后,再感染WI-38人胚肺成纤维细胞,均用ELISA法检测VEGF、Ang1目的基因的表达,并采用鸡胚尿囊膜血管形成实验(CAM)法分析其对血管形成的影响。结果: 扩增的Ad-AV腺病毒效价可达4×10¹⁰pfu/ml;Ad-AV不仅能在293A细胞中成功表达目的基因Ang1、VEGF,而且在WI-38成纤维细胞也能成功表达,其表达产物具有显著的促进CAM上血管生成的活性。结论:成功构建并获得了Ad-Ang1-IRES-VEGF165重组病毒子, 目的基因均能在人胚肾和人胚肺成纤维细胞中表达,其表达产物具有诱导血管形成的功能。     

Selective method for the determination of cefdinir in human plasma using liquid chromatography electrospray ionization tandam mass spectrometry

A sensitive and selective liquid chromatographic-tandem mass spectrometric (LC-MS/MS) method was developed for the determination of cefdinir in human plasma. After a simple protein precipitation using trichloracetic acid, the post-treatment samples were applied to a prepacked RP18 Waters SymmetryShield column interfaced with a triple quadrupole tandem mass spectrometer. Positive electrospray ionization was employed as the ionization source. The mobile phase consisted of methanol-water-formic acid (25:75:0.075, v/v/v). The analyte and I.S. cefaclor were both detected by the use of selected reaction monitoring mode. The method was linear in the concentration range of 5-2,000 ng/ml. The lower limit of quantification was 5 ng/ml. The intra- and inter-day relative standard deviation across three validation runs over the entire concentration range was less than 4.3%. The accuracy determined at three concentrations (36, 360 and 1,800 ng/ml for cefdinir) ranged from 99.6 to 106.7% in terms of recovery. The chromatographic run time for each plasma sample was less than 3 min. The method herein described was successfully applied for the evaluation of pharmacokinetic profiles of cefdinir capsule in 12 healthy volunteers.     

双歧异黄酮奶粉对化疗荷瘤小鼠减毒增效作用的研究

目的：研究双歧异黄酮奶粉对化疗后荷瘤小鼠免疫功能的影响。方法：以S180荷瘤C57BL／6小鼠为模型,系统地研究双歧异黄酮奶粉对瘤重和细胞功能的影响。结果：双歧异黄酮奶粉对荷瘤鼠化疗所引起的免疫功能低下具有明显的恢复作用,可显著提高化疗荷瘤鼠T细胞转化能力,明显促进化疗荷瘤鼠白介素—2(IL—2)分泌水平。结论;双歧异黄酮奶粉能够拮抗肿瘤抗原和化疗所引起的免疫抑制。     

冬小麦光合特征及叶绿素含量对保水剂和氮肥的响应

以不施保水剂和氮（N）肥为对照,测定了保水剂（60 kg·hm^-2）与不同N肥水平（0、225、450 kg·hm^-2）及其配施条件下大田小麦的光合特征、叶绿素含量和水分利用效率等指标,研究了冬小麦拔节期和灌浆期光合生理特征、叶绿素含量及水分利用对保水剂和N肥的响应.结果表明：灌浆期各处理的光合速率、气孔导度、胞间CO₂浓度、叶片水分利用效率及叶绿素含量均大于拔节期.在拔节期,单施N肥条件下,随施N量的增加,单叶水分利用效率提高,光合速率、气孔导度、胞间CO₂浓度及蒸腾速率均先增后减;225 kg·hm^-2 N肥处理的叶绿素含量最高.施用保水剂后,随施N量的增加,胞间CO₂浓度降低,而光合速率等均提高;单施保水剂及其与N肥配施提高了叶绿素含量,而过多N肥效果不显著在灌浆期,单施N肥显著提高了小麦的光合速率及水分利用效率,降低了气孔导度、胞间CO₂浓度及蒸腾速率;叶绿素含量随N肥用量的增加而增加.施用保水剂后,随N肥用量的增加,光合速率和叶片水分利用效率均先增后减,而胞间CO₂浓度和蒸腾速率先减后增,但均低于对照,气孔导度随施N量的增加而提高.单施保水剂的叶绿素含量显著提高,但其与N肥配施叶绿素含量有所降低.保水剂与N肥配合施用显著提高了小麦的千粒重、产量及水分生产效率.其中,保水剂与225 kg·hm^-2N肥配施处理的产量及水分生产效率均最高.     

黑麦1R染色体的显微分离、体外扩增及扩增产物的鉴定

借助Leitz显微操作器,在国产倒置显微镜下(400×)用玻璃针对处于有丝分裂中期的黑麦根尖细胞中的1R染色体成功地进行了分离。分离出来的1R染色体转入0.5 ml的Eppendorf管中,用蛋白酶K处理,把DNA释放出来;经Sau3A酶切,再与人工合成的Sau3A连接头连接;以连接头的一条链的核苷酸顺序片段为引物对DNA酶切片段进行了PCR扩增。琼脂糖凝胶电泳显示扩增产物的长度大约为300~1000 bp。以生物素分别标记的黑麦总体DNA和小麦rDNA为探针进行斑点杂交,结果表明PCR扩增产物确实来源于黑麦的1R染色体DNA。这个方法为构建黑麦1R染色体亚基因组文库和筛选1R染色体特异性探针奠定了基础。     

保水剂用量对小麦不同生育期根系生理特性的影响

在河南禹州试验基地进行田间试验,研究了保水剂不同施用量（0、30、60、90 kg·hm^-2）对两个冬小麦品种（郑麦9694和矮抗58）根系生理生化特征、生物量及产量的影响,以探明保水剂对不同生育阶段冬小麦根系的作用机理.结果表明：施用保水剂降低了冬小麦根系质膜透性和可溶性糖含量,提高了根系活力.在各生育期,郑麦9694根系质膜透性降低幅度均大于矮抗58;除90 kg·hm^-2处理外,矮抗58的根系活力均显著大于郑麦9694.在孕穗期和灌浆期,郑麦9694的可溶性糖含量降低幅度显著大于矮抗58.在各生育期内,60 kg·hm^-2处理的两品种质膜透性和可溶性糖含量均最小,90 kg·hm^-2与60 kg·hm^-2处理差异不显著.随保水剂用量的增加,郑麦9694的根系活力显著提高,而矮抗58 在60 kg·hm^-2处理下最高.施用保水剂还提高了小麦根系生物量,在拔节期和孕穗期,矮抗58的根系生物量大于郑麦9694;而在灌浆期,矮抗58在60 kg·hm^-2和90 kg·hm^-2处理下根系生物量显著低于郑麦9694.郑麦9694和矮抗58产量均以60 kg·hm^-2处理增幅最高.综上,保水剂对郑麦9694的作用效果较矮抗58显著,并以60 kg·hm^-2施用量为佳.