长尖对齿藓原丝体快速培育的关键影响因子研究

为了给苔藓结皮野外恢复的大规模接种提供丰富种源,本文利用0.1% NaClO（10、15、20、30、45、60、120 s）和0.1% HgCl₂（10、15、20、30、45、60、120 s）两种消毒液,采用Knop、MS和Hoagland 3种培养基,设置5.5、6、6.5、7、7.5、8、8.5七个水平的pH值,通过单因素试验设计方法,测定长尖对齿藓[Didymodon ditrichoides（Broth.）]茎叶体的成活率、原丝体长度和分枝数等指标,探讨影响长尖对齿藓原丝体扩繁的关键因子。结果表明：（1）消毒方式、培养基及pH均对长尖对齿藓茎叶体的成活率、原丝体长度和分枝数有显著影响（P<0.05）,影响大小依次均为消毒方式 > 培养基 > pH;（2）最适合的消毒方式为：0.1% NaClO消毒15~20 s;（3）最适合长尖对齿藓原丝体生长的培养基是Knop和Hoagland培养基;（4）最适合长尖对齿藓原丝体生长的pH是7.5。从而得出快速培育长尖对齿藓原丝体的最佳组合为：0.1% NaClO消毒15~20 s+Hoagland/Knop+pH=7.5。本文的研究结果可以缩短长尖对齿藓生长周期以进行快速繁殖,为苔藓结皮的快速培育以及工程化应用提供一定的借鉴。     

补肾温阳化瘀法对子宫内膜异位症患者子宫内膜腺上皮细胞OPN与MMP-9的影响

目的:探讨补肾温阳化瘀法对子宫内膜异位症患者子宫内膜腺上皮细胞骨桥蛋白(OPN)与基质金属蛋白酶-9(MMP-9)的影响。方法:选择我院2013年4月~2016年4月收治的92例子宫内膜异位症患者,分为对照组与观察组,各46例,对照组行常规西医治疗,观察组行补肾温阳化瘀法治疗,比较两组OPN及MMP-9,雌二醇(E2)、促黄体生成素(LH)、卵泡刺激素(FSH),肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、白细胞介素-8(IL-8),CD3~+、CD4~+、CD8~+、CD4~+/CD8~+,临床疗效及安全性。结果:治疗后,观察组OPN、MMP-9,TNF-α、IL-6、IL-8,CD8~+低于对照组,观察组CD3~+、CD4~+、CD4~+/CD8~+高于对照组,差异有统计学意义(P0.05)。观察组总有效率高于对照组,不良反应率低于对照组(P0.05)。结论:补肾温阳化瘀法可下调子宫内膜异位症患者子宫内膜腺上皮细胞OPN及MPP-9表达。     

基于标准病例的案例教学法在普外科见习教学中的应用探讨

目的:探讨基于标准病例(SP)的案例教学法(CBL)在普外科见习教学中的应用效果。方法:选取2014年6月至2015年6月我院见习的五年制临床本科医学生共112人,利用随机数字表的方法将其分为对照组和研究组,两组人数各56人。对研究组采取基于SP的CBL教学法,而对照组则使用传统临床带教方法,1个月后对理论、技能进行评价,并对学生和教师的满意进行问卷调查。结果:在技能考核和病例分析方面,研究组学生的得分显著高于对照组(P0.05),理论成绩无明显差异(P0.05)。对学生满意度的调查中,在"课堂气氛活跃"、"对学习兴趣的激发"、"利于理论与实践结合"以及"有利于形成正确临床思维"四个方面,研究组的满意度显著高于对照组(P0.05)。在对教师满意度调查中,认为试验组教学法的效率要高于对照组教学法(P0.05),而在"易实施性"满意度低于对照组教学法(P0.05),在"可操作性"方面差异无统计学意义(P0.05)。结论:与传统教学模式比较,基于SP的CBL教学法具有显著的优势,能更直观的展示某种疾病的特征,使学生通过实践巩固对基础知识的记忆,值得进一步探索并推广。     

载HCPT纳米粒的制备及其体外抗肿瘤活性研究

目的:制备载羟基喜树碱(HCPT)的PLGA-hyd-PEG-FA纳米粒(HCPT@PLGA-hyd-PEG-FA),并对其体外抗肿瘤活性进行研究。方法:采用乳化溶剂挥发法制备HCPT@PLGA-hyd-PEG-FA,通过单因素试验考察超声功率、聚合物浓度、PVA浓度、水相和油相体积比及投药量对纳米粒粒径的影响;采用zeta电位及激光粒度分析仪测定纳米粒的粒径及zeta电位,用透射电镜(TEM)观察其形态;采用透析法评价HCPT@PLGA-hyd-PEG-FA的体外释药特性;采用MTT法测定HCPT@PLGA-hyd-PEG-FA对HepG2细胞的细胞毒性。结果:HCPT@PLGA-hyd-PEG-FA平均粒径约为109±3 nm,zeta电位为-11.57 mV,载药量为5.6%,TEM显示其为球形;体外释药结果表明HCPT@PLGA-hyd-PEG-FA对HCPT的释放具有p H值依赖性;HCPT和HCPT@PLGA-hyd-PEG-FA的IC50值分别为474.6 ng/mL和286.0 ng/mL。结论:HCPT@PLGA-hyd-PEG-FA体外释药性能良好,HCPT@PLGA-hyd-PEG-FA的细胞毒性明显大于游离的HCPT,值得进一步研究。     

Performance of Holstein calves receiving equal quantities of milk at fixed or variable amounts per day during milk-feeding period

Two experiments were conducted to determine the effects of feeding equal quantities of milk during the pre-weaning period through different milk-feeding regimes on calf growth, starter intake and selected blood metabolites. In experiment 1, 44 female Holstein calves (3 days of age and 39.2±4.3 kg of BW) were distributed randomly to one of two milk-feeding programs (1 calf per pen; 22 pens per treatment group): (1) consistent (CONS; 6 l/day of milk from days 3 to 60 and 3 l/day from days 61 to 65 of age) or (2) step-up/step-down (SUSD; 5 l/day of milk from days 3 to 15, 8 l/day from days 16 to 40, 6 l/day from days 41 to 50, 3 l/day from days 51 to 60 and then 2 l/day from days 61 to 65 of age). No difference between treatments was observed in starter consumption, feed efficiency, hip width and heart girth. However, pre-weaning average daily gain (ADG) tended to be greater in CONS than in SUSD calves (0.78 v. 0.70 kg/day; P=0.07). Blood β-hydroxybutyrate at day 45 (pre-weaning) was lower in SUSD than in CONS calves (0.14 v. 0.21±0.013 mmol/l). In experiment 2, 26 male Holstein calves (3 days of age and 39.4±4.1 kg of BW) were assigned at random to one of two milk-feeding protocols (1 calf per pen; 13 pens per treatment group): (1) consistent (CONS; (7 l/day of milk from days 3 to 40 and 2 l/day from days 41 to 45 of age) or (2) step-down (STD; 8 l/day of milk from days 3 to 30, 4 l/day from days 31 to 40 and 2 l/day from days 41 to 45 of age). The milk-feeding program had no effect on the performance measurements, with the exception that ADG (days 15 to 30), starter intake (days 30 to 45) and heart girth (day 45) were greater in STD than in CONS calves. In conclusion, it appears that if the total amount of milk intake is held constant over the course of milk-feeding period, the method of milk feeding would have negligible effects on calf performance.     

Measurement of pO2 by luminescence lifetime spectroscopy: A comparative study of the phototoxicity and sensitivity of [Ru(Phen)3]2+ and PdTCPP in vivo

Dysfunctions in tissue metabolism can be detected at early stages by oxygen partial pressure (pO₂) measurement. The measurement of emission lifetimes offers very promising and non‐invasive approach to estimate pO₂ in vivo. This study compares two extensively used oxygen sensors and assesses their in vivo oxygen sensitivity and phototoxic effect. Luminescence lifetime of Ru‐polypyridyl complex and of Pd‐porphyrin is measured in the Chick's Chorioallantoic Membrane (CAM) model with a dedicated optical fiber‐based, time‐resolved spectrometer. The Pd‐porphyrin luminescence lifetimes measured in the CAM model exposed to different pO₂ levels are longer and have a broader dynamic range (10–100 μs) than those of Ru‐polypyridyl complex (0.6–1 μs). The combined statistical analysis based on an estimate of the kurtosis and skewness, bootstrapping method and routine normality tests is performed. The indicators of the averages and signal to noise ratio stability are also calculated. The combination of several data processing allows selection of the better sensor for a given application. In particular, it is found that the advantage of Ru‐polypyridyl complex over Pd‐porphyrin is two‐fold: i) Ru‐polypyridyl complex datasets have consistently better statistical characteristics, ii) Ru‐polypyridyl exhibits lower cytotoxicity.

     

Four new limonoids from the seeds of Chukrasia tabularis A. Juss.

Two new C-15 enolic acyl phragmalin-type limonoid orthoesters (1-2) which possessed a C-15-propionyl phragmalin skeleton and two new mexicanolide-type limonoids (3-4) were isolated from the ethanol extract of seeds of Chukrasia tabularis A. Juss. Their structures were established on the basis of spectroscopic analyses and electronic circular dichroism (ECD) exciton chirality method. Additionally, all of the compounds were screened against three human tumor cell lines MCF-7, SMMC-7721, and U2OS.     

A Quadrupole Dalton-based multi-attribute method for product characterization,process development,and quality control of therapeutic proteins

During manufacturing and storage process, therapeutic proteins are subject to various post-translational modifications (PTMs), such as isomerization, deamidation, oxidation, disulfide bond modifications and glycosylation. Certain PTMs may affect bioactivity, stability or pharmacokinetics and pharmacodynamics profile and are therefore classified as potential critical quality attributes (pCQAs). Identifying, monitoring and controlling these PTMs are usually key elements of the Quality by Design (QbD) approach. Traditionally, multiple analytical methods are utilized for these purposes, which is time consuming and costly. In recent years, multi-attribute monitoring methods have been developed in the biopharmaceutical industry. However, these methods combine high-end mass spectrometry with complicated data analysis software, which could pose difficulty when implementing in a quality control (QC) environment. Here we report a multi-attribute method (MAM) using a Quadrupole Dalton (QDa) mass detector to selectively monitor and quantitate PTMs in a therapeutic monoclonal antibody. The result output from the QDa-based MAM is straightforward and automatic. Evaluation results indicate this method provides comparable results to the traditional assays. To ensure future application in the QC environment, this method was qualified according to the International Conference on Harmonization (ICH) guideline and applied in the characterization of drug substance and stability samples. The QDa-based MAM is shown to be an extremely useful tool for product and process characterization studies that facilitates facile understanding of process impact on multiple quality attributes, while being QC friendly and cost-effective.     

Optimization‐based framework for resin selection strategies in biopharmaceutical purification process development

This work addresses rapid resin selection for integrated chromatographic separations when conducted as part of a high‐throughput screening exercise during the early stages of purification process development. An optimization‐based decision support framework is proposed to process the data generated from microscale experiments to identify the best resins to maximize key performance metrics for a biopharmaceutical manufacturing process, such as yield and purity. A multiobjective mixed integer nonlinear programming model is developed and solved using the ε‐constraint method. Dinkelbach's algorithm is used to solve the resulting mixed integer linear fractional programming model. The proposed framework is successfully applied to an industrial case study of a process to purify recombinant Fc Fusion protein from low molecular weight and high molecular weight product related impurities, involving two chromatographic steps with eight and three candidate resins for each step, respectively. The computational results show the advantage of the proposed framework in terms of computational efficiency and flexibility. © 2017 The Authors Biotechnology Progress published by Wiley Periodicals, Inc. on behalf of American Institute of Chemical Engineers Biotechnol. Prog., 33:1116–1126, 2017