注射用转移因子不同生产方法比较

比较从猪脾中提取活性转移因子的不同生产方法,以建立适合产业化的制造工艺。将猪脾匀浆后,分别冻融0、3、5次,再以超滤法或Lawence法提取转移因子。与lawrence法相比,经5次冻融并超滤制备的产品收率高,符合国家标准。匀浆冻融次数对转移因子的收率和质量有一定的影响,超滤法适合于转移因子的大规模生产。     

苦瓜皂苷提取工艺研究

采用正交设计研究回流法苦瓜皂苷的提取工艺。考察提取温度、乙醇浓度、固液比和提取时间等因素在不同水平下对苦瓜皂苷提取率的影响,并对结果进行方差分析和显著性检验。结果显示乙醇提取法最佳工艺组合为A3B1C2D3,即70％乙醇,固液比为1：15,100℃回流提取1．5h,重复提取3次。     

微波法提取枸杞叶甜菜碱工艺的研究

研究微波法提取枸杞叶中甜菜碱的最佳工艺,考察工艺参数对枸杞叶中甜菜碱提取率的影响.以甜菜碱提取率为指标,通过L9(3(4))正交实验与方差分析优选出枸杞叶中甜菜碱最佳提取工艺条件.结果表明,最佳工艺为固液比为(g:ml)1:10,20 min,微波功率200 w.枸杞叶中甜菜碱最佳提取率平均值可高达4.48%.该工艺简...     

正交试验法研究决明子提取工艺

采用正交试验法优选出了决明子中蒽醌衍生物的最佳提取工艺条件为：50％乙醇水为浸提剂;固液比1：5,提取温度70℃,提取时间2h。总蒽醌衍生物的产率为0．83％。如果采用醋酸酸化预处理工艺,可使蒽醌衍生物(抗癌、抗菌有效部位)的产率增加0．19％。通过溶剂萃取和沉淀分离得到两大有效部位：脂溶性部位(蒽醌衍性物)和水溶性部位(蒽醌甙类)。     

以豆腐柴叶为原料提取果胶工艺研究

本文开发了一条从豆腐柴叶中提取果胶的新工艺。该工艺中,树脂吸附纯化、超滤浓缩和喷雾干燥是三个重要的单元操作。为了确定最佳提取条件和考察树脂吸附纯化和超滤浓缩两个单元操作的商业应用可行性,进行了三种不同规模的试验。结果表明,所开发的工艺在能耗和果胶质量方面明显优于传统的醇沉淀法。     

紫花前胡的提取工艺优化研究

目的:采用多指标综合评分法结合响应面法优选紫花前胡香豆素类成分的最佳提取工艺参数.方法:通过HPLC检测紫花前胡中的化学成分伞形花内酯、紫花前胡苷、补骨脂素、花椒毒素和佛手柑内酯的含量,以综合评分值作为评价指标,并对液料比、甲醇浓度和超声时间进行考察,且在单因素试验基础上,运用Box-Behnken设计响应面法优化提取...     

水蒸气蒸馏法提取大蒜挥发油的工艺研究

以挥发油提取率为指标,通过单因素和L9(34)正交试验设计确定了水蒸气蒸馏法提取大蒜挥发油的最佳提取工艺及条件。结果表明,各因素作用主次顺序为蒸馏时间(D)>料液比(C)>发酵温度(A)>浸泡时间(B),各因子的最优组合为A2B2C2D3,即以1∶3.5的料液比(m∶V),在35℃时浸泡3h后蒸馏2h为大蒜挥发油的最佳提取工艺,此时挥发油的提取率达到0.32%,证明此方法是大蒜挥发油提取的可靠、易行的科学方法。     

罗汉果皂甙的提取工艺研究

从罗汉果中提取了具有保健作用的强甜味的皂甙。实验证明Ｃａ（ＯＨ）２是一种很好的澄清剂,Ｄ２８０、Ｄ２９０、Ｄ６１和Ｄ１５１三种国产离交树脂有很好的脱色能力,ＡＢ０８吸附树脂能有效地分离这种皂甙。     

茶多酚提取优化工艺研究

本文研究了可食用茶多酚的提取优化工艺。萃取剂为乙醇,料液比1：15,浸提过程伴随300W超声波震荡,就乙醇的浓度、提取温度、提取时间和提取次数等因素利用正交设计筛选了茶多酚的最佳提取工艺条件,并对浸提液最佳离子沉淀方法作了比较。结果表明,茶多酚提取的最佳工艺条件为：65％的乙醇溶液作浸提剂,当浸提温度为50℃,浸提时间30min,两次超声波辐射浸提后茶多酚从粗茶叶中的提取率为20．1％。沉淀剂AlCl3和ZnSO4质量比为1：2对茶多酚的沉淀效果最佳,pH=6．0。     

肺传递因数测试方法的研究

本文介绍了一种有于测试肺的传递因数的仪器,探讨了改进设计的方法和影响传递因数的一些因素。在仪器设计中使用了随机参数的贝叶斯计算技术来估计肺的传递因数。实验结果表明这个参数与生物统计参数之间有较强的关系,而且肯定了特色文献的发现,即测量持续时间可以缩短。     

Nucleoplasmin facilitates reprogramming and in vivo development of bovine nuclear transfer embryos

Successful cloning by somatic cell nuclear transfer (NT) involves an oocyte-driven transition in gene expression from an inherited somatic pattern, to an embryonic form, during early development. This reprogramming of gene expression is thought to require the remodeling of somatic chromatin and as such, faulty and/or incomplete chromatin remodeling may contribute to the aberrant gene expression and abnormal development observed in NT embryos. We used a novel approach to supplement the oocyte with chromatin remodeling factors and determined the impact of these molecules on gene expression and development of bovine NT embryos. Nucleoplasmin (NPL) or polyglutamic acid (PGA) was injected into bovine oocytes at different concentrations, either before (pre-NT) or after (post-NT) NT. Pre-implantation embryos were then transferred to bovine recipients to assess in vivo development. Microinjection of remodeling factors resulted in apparent differences in the rate of blastocyst development and in pregnancy initiation rates in both NPL- and PGA-injected embryos, and these differences were dependent on factor concentration and/or the time of injection. Post-NT NPL-injected embryos that produced the highest rate of pregnancy also demonstrated differentially expressed genes relative to pre-NT NPL embryos and control NT embryos, both of which had lower pregnancy rates. Over 200 genes were upregulated following post-NT NPL injection. Several of these genes were previously shown to be downregulated in NT embryos when compared to bovine IVF embryos. These data suggest that addition of chromatin remodeling factors to the oocyte may improve development of NT embryos by facilitating reprogramming of the somatic nucleus.     

Radioimmunoassay for fibroblast growth factor (FGF): release by the bovine anterior pituitary in vitro

A radioimmunoassay (RIA) was developed to measure fibroblast growth factor (FGF) using antiserum generated against a synthetic replicate of [Tyr¹⁰]FGF(1–10). The antisera, previously shown to be capable of inhibiting the biological action of FGF on bovine aortic arch endothelial cells in vitro [1], are highly specific for the amino-terminus of FGF. In the RIA, the antisera recognize the decapeptide antigen [Tyr¹⁰]FGF(1–10) and the intact mitogen on an equimolar basis and show less than 0.01% cross-reactivity with N-acetyl-[Tyr¹⁰]FGF(1–10).

Bovine adenohypophysial cells maintained in primary monolayer culture release and ir-FGF which is indistinguishable from the intact mitogen in as much as it is retained on heparin-Sepharose affinity columns and shows a dose-dependent and parallel displacement in RIA. The release of ir-FGF by the bovine adenohypophysis can be increased with forskolin (10⁻⁵ M) or KCl (50 mM). Preincubation of pituitary cells with 17β-estradiol has no measurable effects on basal ir-FGF, but increases the release after KCl treatment 2–3-fold. These results show that ir-FGF can be released by the bovine adenohypophysis in vitro and lend credence to the hypothesis that FGF plays a physiological role in the homeostatic mechanisms regulating mesoderm-derived cell growth.     

细胞核移植牛供质卵母细胞研究现状

核移植牛的研究具有巨大的经济价值,国外对此的研究不断深入,但国内开展此项研究相对滞后。本文就牛卵母细胞的成熟、去核与激活作一综述,重点介绍目前国外常用的方法,包括其效率和影响因素,其中涉及到一些常用的参数,这对从事核移植牛的同仁们会有一定的参考价值。     

Plasmodium berghei: The spleen in sporozoite-induced immunity to mouse malaria

A/J mice were splenectomized (Splx) or sham-splenectomized (SSplx) prior to administration of a single injection of irradiated sporozoites. Following challenge 7 days after immunization, it was found that none of the splenectomized mice were protected whereas 50% of the sham-splenectomized and intact animals were resistant to challenge. In another series of experiments similar groups, along with mice splenectomized just prior to challenge, were injected with 1.5 × 10⁵ irradiated sporozoites over a 5 week period. This resulted in protection of (1) 60–100% of the animals splenectomized before immunization, and (2) 90–100% protection of the animals splenectomized prior to challenge, as well as the intact and sham-splenectomized mice. Serum levels of antisporozoite antibodies (CSP and SNA) increased during immunization of the intact animals. Only 15–20% of the animals splenectomized prior to immunization presented positive CSP reactions and little if any sporozoite neutralizing activity (SNA) was detected. Serum from intact animals immunized and found resistant to sporozoite challenge was used for passive transfer studies. Immune serum recipients were challenged with small numbers of sporozoites. Only one out of 18 recipients was protected against sporozoite challenge.     

Nucleolar and mitochondrial morphology in bovine embryos reconstructed by nuclear transfer

The nucleolar and mitochondrial morphology of developing reconstructed bovine nuclear transfer (NT) embryos and stage-matched in vivo-produced control embryos were examined under the electron microscope. Each reconstructed embryo at the one-cell (n = 12), two-cell (n = 5), three-cell (n = 3), four-cell (n = 5), 5–8 cell (n = 5) and blastocyst (n = 3) stages was produced by fusion of a 16–32-cell-stage blatomere with an aged enucleated bovine oocyte. The normal and reconstructed embryos showed similar mitochondrial morphology. However, NT embryos produced several pleiomorphic forms not seen in controls, and were more heterogenous at early stages of development. Control embryos exhibited nucleolar features considered indicative of rRNA synthesis from the eight-cell stage onwards. In contrast, the NT embryos presented nucleoli with morphology consistent with rRNA synthesis in all embryos examined, except in the three-cell and in two of the five four-cell embryos. From this nucleolar morphology, it was concluded that nuclear reprogramming does not occur immediately following nuclear transfer, but occurs gradually over the first two or three cell cycles. © 1996 Wiley-Liss, Inc.     

Pilot Study of Bioaccumulation and Distribution of Cesium,Potassium, Sodium and Calcium in King Oyster Mushroom (Pleurotus Eryngii) Grown Under Controlled Conditions

This pilot study presents preliminary results on interrelations between alkali and alkaline earth elements during their transfer to mycelium and fruitbodies of saprophytic fungi. The accumulation and distribution of four elements (cesium, potassium, sodium, and calcium) was evaluated in king oyster mushroom (Pleurotus eryngii) cultivated under controlled conditions. Elemental composition of caps, stipes, and the substrate was analyzed by atomic absorption/emission spectroscopy to evaluate discrimination, concentration, and transfer factors. The transfer factors determined for all the investigated elements were different and can be put in the following order: Cs > K > Na > Ca. There has been a higher accumulation of cesium in caps than in stipes. Distribution of cesium in fruitbodies depended on the presence of other ions in the substrate. The addition of Ca²⁺ limited the transport of cesium and potassium from stipes to caps. Sodium and calcium were mainly accumulated in the stipes. In a control experiment, without supplementation with K⁺, Na⁺, and Ca²⁺, ～ 62% of the cesium present in the substrate was extracted by mycelium and transported to the fruitbodies. Possible applications of fruiting saprophytic fungi in bioremediation are discussed.     

猪脾转移因子对La Sota株鸡新城疫弱毒疫苗的免疫增强作用

【目的】本文旨在研究猪脾转移因子（TF）对新城疫（ND）病毒弱毒疫苗La Sota株的免疫增强作用及机理,为兽医临床防控提供理论依据。【方法】以4种不同剂量La Sota疫苗株分别单独、与TF联合免疫SPF鸡,14 d后以ND病毒（NDV）参考强毒F₄₈E₉株(10^4.7 ELD₅₀)进行攻毒,采用蛋白质芯片技术测定鸡外周血IL-6、IL-10、IL-16和IL-21浓度,并以血凝抑制（HI）试验和荧光定量RT-PCR方法分别检测鸡ND HI抗体效价和F₄₈E₉株的病毒血症水平。【结果】攻毒保护试验表明,单独免疫10^5.17、10^4.17、10^3.17和10^2.17 EID₅₀剂量时疫苗攻毒保护率分别为100%、55%、0%和0%,半数保护量(PD₅₀)为12 023 EID₅₀;对应剂量联合免疫时疫苗攻...     

Effect of two activation treatments and age of blastomere karyoplasts on in vitro development of bovine nuclear transfer embryos

The yield and quality of (a) parthenogenetic blastocysts produced by two activation treatments (cycloheximide [CHX] or 6-dimethylaminopurine [DMAP]) and (b) nuclear transfer blastocysts generated using these two activation treatments and three different ages of karyoplast derived from day 3, 4, or 5 in vitro produced donor embryos, were examined in order to define an optimal nuclear transfer protocol. The two activation protocols comprised calcium ionophore followed by either CHX or DMAP. Parthenogenetic blastocyst yields were greater (P < 0.001) following activation with DMAP than CHX (59.7 +/- 5.1 vs. 31.4 +/- 4.5 [mean +/- SEM]). In contrast, nuclear transfer blastocyst rates per fused embryo were lower (P < 0.0001) using cytoplasts activated with DMAP. The individual rates using day 3, 4, and 5 donors and using CHX and DMAP activation treatments were 31.9 +/- 5.0, 31.7 +/- 6.2, 20.4 +/- 7.3 and 27.8 +/- 4.7, 20.1 +/- 7.5, 12.7 +/- 8.3, respectively. Blastocyst rate per fused embryo was negatively correlated (P = 0.0091) with the total number of blastomeres per donor embryo. Despite this inverse relationship, the calculated potential blastocyst yield per donor embryo was positively correlated (P < 0.0048) to karyoplast age. The individual potential yields on days 3, 4, and 5 and for the two activation protocols (CHX and DMAP) were 4.7 +/- 0.8, 7.2 +/- 1.2, 10.1 +/- 2.1 and 3.8 +/- 0.8, 5.5 +/- 2.1, 7.3 +/- 4.1, respectively. One possible explanation for the observed inverse relationship is that differentiation events during early cleavage are able to reduce the ability of the cytoplast to reprogram the transferred karyoplast and hence reduce blastocyst yields. The mechanism that mediates the differential effect of the CHX and DMAP on blastocysts yields between parthenogenetic and nuclear transfer embryos remains to be elucidated. In conclusion, the results indicate that although activation of oocytes with DMAP can produce a higher percentage of blastocysts, CHX activation is superior for use in nuclear transfer.