树莓叶黄酮提取的正交试验和响应曲面法工艺优化对比

旨在为树莓叶黄酮的开发应用提供数据参考,通过正交试验与响应曲面法设计改良乙醇提取工艺,并通过HPLC建立树莓叶黄酮的色谱分离条件为:Agilent ZorBax SB色谱柱,流动相为1mg/mL乙酸-乙腈=72.5∶27.5,等度分离,柱温30℃,检测波长:257nm,完成树莓叶黄酮的初步分离,以及黄酮中芦丁含量的准确测定和验证。最佳提取工艺为:646mL/mL乙醇,料液比1∶12.5,41.9℃加热回流173.3min,最佳工艺所得提取率1.127%,响应曲面法最佳工艺提取率略高于正交试验法,且其工艺条件均较优于正交试验所得工艺,可为后期研发树莓叶芦丁兽用中药制剂提供理论参考。     

响应面法优化当归补血汤液体发酵工艺

为确定当归补血汤液体发酵的最佳工艺,本试验采用枯草芽孢杆菌为发酵菌种,以多糖提取率为评价指标进行液体发酵工艺优化。首先采用单因素试验考察发酵培养基中葡萄糖和蛋白胨添加量、菌液接种量和发酵时间对多糖提取率的影响,再以此为基础,采用Box-Behnken中心组合试验设计法,选取葡萄糖添加量、蛋白胨添加量、接种量、发酵时间为发酵优化的4个因素,以多糖提取率为响应值,设计4因素3水平的试验方案并进行响应面分析,以确定各因素与响应值多糖提取率的关系。结果显示,各因素对多糖提取率变化的贡献大小依次为葡萄糖添加量、蛋白胨添加量、接种量、发酵时间;利用响应面分析结果显示,蛋白胨添加量与接种量和发酵时间、接种量和发酵时间的交互作用较为显著,最终确定最佳发酵工艺条件为：葡萄糖添加量0.95%,蛋白胨添加量1.52%,接种量2.92%,发酵时间35.8 h。该条件下多糖提取率达到9.23%,与未发酵组（4.62%）相比,多糖含量提高近2倍。该工艺对液体发酵当归补血汤产多糖优化效果显著,为发酵当归补血汤的进一步开发利用提供了理论基础。     

野菊和神农香菊毛状体及叶片表面分泌物的比较

本研究以野菊(Dendranthema indicum)和神农香菊(D.indicumvar.aromaticum)为研究对象,对比分析了其叶片的表皮毛形态、密度以及叶片表面分泌物种类和含量。结果表明,其叶片表皮毛主要有两种类型,一类是没有分泌功能的T-形非腺毛,一类是具有分泌功能的头状腺毛。T-形非腺毛在上下表皮的密度差异不大,而头状腺毛差异较大,主要集中在叶片的下表皮。两种植物材料叶片表面分泌物里共分析出73种化合物,其中大部分为萜类及其衍生物,共有的化合物有18种;野菊和神农香菊中分别分析出48、43种化合物,分别占分泌物总相对含量的90.96%和84.33%。     

Characterization of tuberculous granulomas in different stages of progression and associated tertiary lymphoid tissue in goats experimentally infected with Mycobacterium avium subsp. hominissuis

Oral infection of goats with Mycobacterium avium subsp. hominissuis (MAH) resulted in a large variety of granulomas in organized gut-associated lymphatic tissues and intestinal lymph nodes. To characterize the cellular composition of granulomas, CD4⁺, CD8⁺, γδ, B lymphocytes and plasma, CD25⁺, CD68⁺, MHC-II⁺, Ki67⁺ and endothelial cells were labeled in consecutive frozen sections by immunohistochemistry and acid fast bacilli (AFB) by Kinyoun stain. Granulomas with extensive necrosis, little mineralization and variable numbers of AFB surrounded by many CD4⁺ T cells, but only few epitheloid macrophages were observed in severely sick goats at 2–3 mpi. They were interpreted as exuberant immune reaction. Organized granulomas with very few AFB were seen in clinically healthy goats at 13 mpi. The necrotic cores were surrounded by a zone of granulomatous infiltrate with many epitheloid macrophages and few lymphocytes. This zone was initially wide and highly vascularized and became progressively smaller. It was enclosed by an increasing layer of connective tissue. All organized granulomas were surrounded by compartimentalized tertiary lymphoid tissue. The granulomas in experimental infection of goats with MAH reflect the heterogeneity of lesions seen in mycobacterial infections of humans and ruminants and are therefore valuable for comparative research.     

3个棘孢木霉菌株对山新杨组培移栽苗生长和光合特性的影响

为了收集具有促进林木生长作用的棘孢木霉(Trichoderma asperellum)功能菌株,本研究采用盆栽方法,以1年龄山新杨(Populus davidiana×P.albavar.pyramidalis)组培移栽苗为试材,分析了根施5×10~3 cfu·cm~(-3)土的3个棘孢木霉菌株Ta492、Ta536和Ta4分生孢子对山新杨幼苗生长状况、叶绿素含量、光合日变化及光响应曲线特征参数的影响。结果发现,经3个棘孢木霉菌株诱导后,杨树苗苗高、地径及生物量均不同程度地高于对照组(CK);杨树苗叶绿素含量和叶绿素a/b在多数时间点均显著高于CK(P0.05),为光合速率的提高奠定了物质基础。同时,试验组杨树苗叶片的净光合速率(P_n)、气孔导度(G_s)、蒸腾速率(T_r)的日变化均高于CK,但变化趋势均与CK相同;并且,最大净光合速率(P_(n max))、光饱和点(LSP)、表观量子效率(AQY)等光响应曲线特征参数也在不同程度上高于CK。说明这3个棘孢木霉菌株均能有效提高山新杨的光合能力,促进其生长。并且,Ta536菌株对山新杨幼苗的诱导效果最好。