猪链球菌蛋白表面展示系统的构建及展示蛋白的初步观察

为构建猪链球菌(Streptococcus suis)蛋白表面展示系统,本研究通过序列分析,确定猪链球菌的LPxTG蛋白及其信号肽(SP)和胞壁锚定基序(CWA),通过PCR扩增Peno-SP、GFP、CWA的DNA片段并融合,构建强启动子Peno控制表达编码SP-GFP-CWA融合蛋白的DNA片段,将该重组DNA片段连接pSET2载体,获得蛋白表面展示质粒,转化猪链球菌,构建得到以GFP为报告蛋白的猪链球菌蛋白表面展示系统。结果显示,利用猪链球菌的10个LPxTG蛋白及其SP和CWA序列,构建了10个含有Peno-SP-GFP-CWA融合片段的重组pSET2表面蛋白展示质粒pSsPSD1至pSsPSD10,分别转化猪链球菌05ZYH33,PCR鉴定显示其中7个转化猪链球菌。采用western blot初步检测其展示蛋白,结果显示,7个转化阳性菌株均能有效表达GFP蛋白,以成熟GFP条带为指标,均表现出了一定的外源GFP表面展示水平,分别命名为SsPSD1、SsPSD2、SsPSD4、SsPSD7-SsPSD10,其中SsPSD1、SsPSD4、SsPSD8和SsPSD9表面展示水平相对较好,在猪链球菌表面展示外源蛋白方面具有很好的潜力。本研究首次尝试建立猪链球菌蛋白表面展示系统,为猪链球菌表面递呈外源蛋白或抗原提供了新的策略。     

Cigarette dust particles induced lung function injury of chronic obstructive pulmonary disease in mice

AIM: To investigate the pathogenesis of chronic obstructive disease (COPD) in mice by using nasal drip of cigarette dust particles (DSP) induced pulmonary function damage model.METHODS: BALB/c mice were randomly divided into control group, LPS 100 mg/L group, DSP 0.75 mL/L group, DSP 1.5 mL/L group and DSP 3 mL/L group for 30 days. The method of nasal drip was used for 30 days to establish the COPD model. Rrs, Ers, Crs, Est, Cst, P-3/8Rn, P-3/8G and P-3/8H were measured for evaluating lung function of the mice in each group by the method of FlexiVent. The effect on the increase of airway resistance induced by methacholine (Mach) was determined using main bronchial rings by Myograph method. The HE, Masson and Resorcinol fuchsin staining of mouse tracheas and lung tissues were conducted. RESULTS: Continuous nasal drip with DSP for 30 days increased Rrs, Ers, Est, P-3/8Rn and decreased Crs, Cst, P-3/8G and P-3/8H in the mice. DSP significantly shifted the dose-effect curve of tracheal contraction induced by Mach to the left, increased the sensitivity of the airway to Mach, and significantly increased the maximal contractile airway effect of Mach. Exposure to DSP caused fibrosis of airway subepithelial, deposition of collagen in the airway basement membrane under the reticular plate, induced reticular plate thickening, pulmonary bronchial lumen serious deformation, and the inflammatory cell infiltration in the lung of mice. Significantly increased alveolar wall muscle fibers and collagen fibers were also observed. CONCLUSION: The lung function and pathomorphological changes of COPD mice induced by 30 days nasal drip of cigarette dust particles were similar to those of human COPD.     

大球盖菇1号的分离纯化及其液体培养基优化

为缩短大球盖菇(Stropharia rugoso-annulata)1号液体种生长周期,提高菌丝体产量,为大球盖菇液体种工业化生产提供参考和依据。试验以大球盖菇1号菌丝球干质量(Y)为指标,通过Plackett-Burman设计试验,得出玉米粉、MgSO4和VB1对大球盖菇的生物量有显著影响,通过最陡爬坡试验和Box-Behnken响应面分析法对大球盖菇液体培养基组成进行优化,得到大球盖菇1号最适液体培养基为蔗糖20 g·L^-1、蛋白胨6 g·L^-1、酵母膏2 g·L^-1、KH2PO43 g·L^-1、MgSO44.56 g·L^-1、玉米粉4.25 g·L^-1、VB12.43 g·L^-1,通过对模型进行验证试验,大球盖菇1号菌丝生物量为1.2154 g·150-1mL-1,较基础培养基相比生物量提高36.2%,且数学模型的预测值与试验观察值相符,相对误差约为1.9%,证明响应面法优化大球盖菇1号液体培养基可行。     

钢支撑代替混凝土支撑可行性分析

在地铁深基坑施工中，围护结构对基坑的安全起着至关重要的作用。本文依据合肥地铁一号线明光路站，施工过程中，由于车站3号出入口为出土口，原计划的第一道钢筋混凝土支撑无法实现，更改为钢支撑，通过有限元分析软件MIDAS-GTS\NX，建立二维模型进行数值模拟，对围护结构的变形和地表沉降进行分析，结果表明：第一道支撑采用钢支撑代替钢筋混凝土支撑对围护结构的侧移值影响不大，钢筋混凝土支撑控制地表沉降的效果要比钢支撑好，对今后类似工程变更提供依据。     

TFSQ型高效环保组合式粮食杂质清理筛的研制

通过对振动筛理、垂直风选、旋风分离与沉降系统的研究,研制出集振动筛理、风选、旋风分离沉降、含尘气流循环再利用为一体的TFSQ型高效环保组合式粮食杂质清理筛。经应用测试和专家鉴定:该设备具有清理效率高、清理过程粉尘污染小,使用维护方便、可靠性强等优点,具有广阔的市场推广前景。     

温度和盐度对吉富品系尼罗罗非鱼受精率和孵化率的联合影响

采用中心复合设计和响应曲面方法研究了温度(T,℃)和盐度(S)对吉富品系尼罗罗非鱼人工受精率(FR,％)和孵化率(HR,％)的综合影响.设定的温度范围为20～34,盐度范围为0～18.建立了温度、盐度与受精和孵化之间关系的定量模型,并通过分析明确了温度与盐度的最优组合.结果表明,吉富罗非鱼在温度与盐度较高或较低范围内受精率与孵化率都处于较低水平.温度和盐度的一次与二次效应对受精与孵化均有极显著的影响(P＜0.01).温度与盐度二者间协同效应不显著(P＞0.05).建立了吉富罗非鱼受精率与孵化率的多项式方程,复相关系数分别达到了0.957 8和0.960 1 (P＜0.01),可以用于预测吉富罗非鱼的受精率和孵化率.利用统计优化技术明确,在温度27.14℃,盐度8.95时,罗非鱼最佳的受精率为80.8％,孵化率为77.63％.本研究旨在考察温度与盐度同时变化对吉富罗非鱼受精率与孵化率的影响,探讨最佳温度－盐度组合以增加吉富罗非鱼苗种产量.     

Preparation of Adjuvant Diluent for Live Vaccine against Classical Swine Fever and Evaluation of its Immune Effects

The aim of this study was to develop a novel oil-in-water (O/W) emulsion as adjuvant diluents (AD) for live vaccine against classical swine fever (CSF) that could effectively enhance the immune effect of vaccine.The AD was prepared by high-pressure homogenization technique.Formulations and preparation parameters were optimized with response surface design.Its stability, particle size, polydispersity (PDI) and Zeta potential were characterized.The humoral immune response and cellular immune response of the AD were evaluated with BALB/c mice by intramuscular injection.The particle size of the AD prepared by optimized formulation and parameters was 100.4 nm, PDI was 0.147, and Zeta potential was —28.7 mV.The experiment results showed that the AD had good stability.The AD was inoculated combined with live vaccine against CSF into BALB/c mice by intramuscular injection.The results showed that the live vaccine against CSF specific immune responses could be evoked in mice by co-inoculation with AD and vaccine.The cellular immune response levels in co-inoculated groups were significantly higher than control group (P<0.05), with obvious phenomena of higher levels of IFN-γ, IL-6 and IL-4 in serum.The result revealed that cellular immune capability significantly improved with the AD.The results strongly revealed that cellular immune capability significantly improved by introducing AD for effective immune-adjuvant for live vaccine against CSF.     

超声波辅助溶剂法提取金线莲总黄酮工艺优化

旨在研究金线莲总黄酮的最佳提取工艺条件。采用超声波辅助溶剂法,在单因素实验的基础上,采用响应面法优化了金线莲总黄酮的提取工艺。结果表明:当金线莲颗粒度为100目、乙醇体积分数为40%、料液比为1:40(mg:m L)、超声波功率为250 W、超声波时间为30 min时,所得金线莲总黄酮提取率为2.13%。超声波辅助溶剂法强化了金线莲总黄酮的提取效果。     

美味牛肝菌中油脂提取工艺的研究

采用超声波辅助法提取美味牛肝菌中的油脂，在单因素（提取时间、液料比和温度）实验考察的基础上，以油脂提取率为指标，通过响应面法对美味牛肝菌油脂的提取工艺进行优化。实验结果表明：响应面回归模型显著，拟合性好，可用于预测油脂提取率；优化后的油脂提取工艺为：液料比34.2 mL/g，温度36℃，提取时间4.5 h，此条件下油脂提取率为3.54826 %，与模型预测值相符，表明优化的油脂提取工艺合理。